Clinical review: The role of advanced glycation end products in progression and complications of diabetes.
Goh Su-Yen,Cooper Mark E
The Journal of clinical endocrinology and metabolism
CONTEXT:Diabetic complications appear to be multifactorial in origin, but in particular, the biochemical process of advanced glycation, which is accelerated in diabetes as a result of chronic hyperglycemia and increased oxidative stress, has been postulated to play a central role in these disorders. Advanced glycation involves the generation of a heterogenous group of chemical moieties known as advanced glycated end products (AGEs), this reaction occurring as a result of a nonenzymatic reaction with glucose interacting with proteins, lipids, and nucleic acids, and involves key intermediates such as methylglyoxal. EVIDENCE SYNTHESIS:In this review we report on how these AGEs may exert deleterious effects in diabetes, as well as address current strategies to interrupt the formation or action of AGEs. First, AGEs act directly to induce cross-linking of long-lived proteins such as collagen to promote vascular stiffness, and, thus, alter vascular structure and function. Second, AGEs can interact with certain receptors, such as the receptor for AGE, to induce intracellular signaling that leads to enhanced oxidative stress and elaboration of key proinflammatory and prosclerotic cytokines. Over the last decade, a large number of preclinical studies have been performed, targeting the formation and degradation of AGEs, as well as the interaction of these AGEs with receptors such as the receptor for AGE. CONCLUSION:It is hoped that over the next few years, some of these promising therapies will be fully evaluated in the clinical context with the ultimate aim to reduce the major economical and medical burden of diabetes, its vascular complications.
GLUT-1 overexpression: Link between hemodynamic and metabolic factors in glomerular injury?
Gnudi Luigi,Viberti GianCarlo,Raij Leopoldo,Rodriguez Veronica,Burt Davina,Cortes Pedro,Hartley Barry,Thomas Stephen,Maestrini Sabrina,Gruden Gabriella
Hypertension (Dallas, Tex. : 1979)
Mesangial matrix deposition is the hallmark of hypertensive and diabetic glomerulopathy. At similar levels of systemic hypertension, Dahl salt-sensitive but not spontaneously hypertensive rats (SHR) develop glomerular hypertension, which is accompanied by upregulation of transforming growth factor beta1 (TGF-beta1), mesangial matrix expansion, and sclerosis. GLUT-1 is ubiquitously expressed and is the predominant glucose transporter in mesangial cells. In mesangial cells in vitro, GLUT-1 overexpression increases basal glucose transport, resulting in excess fibronectin and collagen production. TGF-beta1 has been shown to upregulate GLUT-1 expression. We demonstrated that in hypertensive Dahl salt-sensitive (S) rats fed 4% NaCl (systolic blood pressure [SBP]: 236+/-9 mm Hg), but not in similarly hypertensive SHR (SBP: 230+/-10 mm Hg) or their normotensive counterparts (Dahl S fed 0.5% NaCl, SBP: 145+/-5 mm Hg; and Wistar-Kyoto, SBP: 137+/-3 mm Hg), there was an 80% upregulation of glomerular GLUT-1 protein expression (P< or =0.03). This was accompanied by a 2.7-fold upregulation of TGF-beta1 protein expression in glomeruli of DSH compared with DSN rats (P=0.02). TGF-beta1 expression was not upregulated and did not differ in the glomeruli of Wistar-Kyoto and SHR rats. As an in vitro surrogate of the in vivo hemodynamic stress imposed by glomerular hypertension, we used mechanical stretching of human and rat mesangial cells. We found that after 33 hours of stretching, mesangial cells overexpressed GLUT-1 (40%) and showed an increase in basal glucose transport of similar magnitude (both P< or =0.01), which could be blocked with an anti TGF-beta1-neutralizing antibody. These studies suggest a novel link between hemodynamic and metabolic factors that may cooperate in inducing progressive glomerular injury in conditions characterized by glomerular hypertension.
High glucose evokes an intrinsic proapoptotic signaling pathway in mesangial cells.
Mishra Rangnath,Emancipator Steven N,Kern Timothy,Simonson Michael S
BACKGROUND:In response to chronic hyperglycemia, microvascular cells undergo stress and injury, which can lead to cell death. We characterized a proapoptotic signaling pathway whereby high glucose evokes an intrinsic, caspase-9-dependent mechanism of cell death in human mesangial cells. METHODS:Biochemical (caspase activity, cytochrome-c release, etc.) and morphologic (chromatin condensation and nuclear segmentation) features of apoptotic cell death were assessed in cultured human mesangial cells exposed to high glucose, a risk factor for mesangial cell injury and diabetic glomerulosclerosis. Proapoptotic signaling was also analyzed in the db/db murine model of kidney injury in diabetes. RESULTS:Incubation in high glucose caused cytotoxicity and apoptosis in mesangial cells. High glucose stimulated mitochondrial release of cytochrome-c, cleavage of procaspase-9, and caspase-9 enzyme activity, suggesting an intrinsic pathway of proapoptotic signaling. In contrast, caspase-8 was unaffected by high glucose. A cell-permeable, caspase-9-selective inhibitor blocked caspase-3 activation and prevented chromatin condensation and nuclear segmentation in cells treated with high glucose. To determine whether an intrinsic signaling pathway occurs in the diabetic kidney in vivo, apoptosis was investigated in diabetic 8- and 16-week db/db murine kidneys. Effector caspases-3 and -7 were activated in diabetic db/db kidneys but not in age-matched nondiabetic db/m controls. At 16 weeks, apoptotic cells in db/db glomeruli were identified on the basis of nuclear segmentation and DNA fragmentation. Apoptosis of glomerular cells correlated with expansion of the mesangial matrix and with worsening of albuminuria. Consistent with an intrinsic signaling pathway, caspase-9 cleavage was elevated only in db/db kidneys, whereas activation of caspase-8 and caspase-12 was undetectable. CONCLUSION:These findings support the hypothesis that hyperglycemia evokes an intrinsic pathway of proapoptotic signaling in mesangial cells. In addition, these results point to an important role for the intrinsic pathway in microvascular injury in the diabetic kidney in vivo.
A metagenome-wide association study of gut microbiota in type 2 diabetes.
Qin Junjie,Li Yingrui,Cai Zhiming,Li Shenghui,Zhu Jianfeng,Zhang Fan,Liang Suisha,Zhang Wenwei,Guan Yuanlin,Shen Dongqian,Peng Yangqing,Zhang Dongya,Jie Zhuye,Wu Wenxian,Qin Youwen,Xue Wenbin,Li Junhua,Han Lingchuan,Lu Donghui,Wu Peixian,Dai Yali,Sun Xiaojuan,Li Zesong,Tang Aifa,Zhong Shilong,Li Xiaoping,Chen Weineng,Xu Ran,Wang Mingbang,Feng Qiang,Gong Meihua,Yu Jing,Zhang Yanyan,Zhang Ming,Hansen Torben,Sanchez Gaston,Raes Jeroen,Falony Gwen,Okuda Shujiro,Almeida Mathieu,LeChatelier Emmanuelle,Renault Pierre,Pons Nicolas,Batto Jean-Michel,Zhang Zhaoxi,Chen Hua,Yang Ruifu,Zheng Weimou,Li Songgang,Yang Huanming,Wang Jian,Ehrlich S Dusko,Nielsen Rasmus,Pedersen Oluf,Kristiansen Karsten,Wang Jun
Assessment and characterization of gut microbiota has become a major research area in human disease, including type 2 diabetes, the most prevalent endocrine disease worldwide. To carry out analysis on gut microbial content in patients with type 2 diabetes, we developed a protocol for a metagenome-wide association study (MGWAS) and undertook a two-stage MGWAS based on deep shotgun sequencing of the gut microbial DNA from 345 Chinese individuals. We identified and validated approximately 60,000 type-2-diabetes-associated markers and established the concept of a metagenomic linkage group, enabling taxonomic species-level analyses. MGWAS analysis showed that patients with type 2 diabetes were characterized by a moderate degree of gut microbial dysbiosis, a decrease in the abundance of some universal butyrate-producing bacteria and an increase in various opportunistic pathogens, as well as an enrichment of other microbial functions conferring sulphate reduction and oxidative stress resistance. An analysis of 23 additional individuals demonstrated that these gut microbial markers might be useful for classifying type 2 diabetes.
Gut microbiota in human adults with type 2 diabetes differs from non-diabetic adults.
Larsen Nadja,Vogensen Finn K,van den Berg Frans W J,Nielsen Dennis Sandris,Andreasen Anne Sofie,Pedersen Bente K,Al-Soud Waleed Abu,Sørensen Søren J,Hansen Lars H,Jakobsen Mogens
BACKGROUND:Recent evidence suggests that there is a link between metabolic diseases and bacterial populations in the gut. The aim of this study was to assess the differences between the composition of the intestinal microbiota in humans with type 2 diabetes and non-diabetic persons as control. METHODS AND FINDINGS:The study included 36 male adults with a broad range of age and body-mass indices (BMIs), among which 18 subjects were diagnosed with diabetes type 2. The fecal bacterial composition was investigated by real-time quantitative PCR (qPCR) and in a subgroup of subjects (N = 20) by tag-encoded amplicon pyrosequencing of the V4 region of the 16S rRNA gene. The proportions of phylum Firmicutes and class Clostridia were significantly reduced in the diabetic group compared to the control group (P = 0.03). Furthermore, the ratios of Bacteroidetes to Firmicutes as well as the ratios of Bacteroides-Prevotella group to C. coccoides-E. rectale group correlated positively and significantly with plasma glucose concentration (P = 0.04) but not with BMIs. Similarly, class Betaproteobacteria was highly enriched in diabetic compared to non-diabetic persons (P = 0.02) and positively correlated with plasma glucose (P = 0.04). CONCLUSIONS:The results of this study indicate that type 2 diabetes in humans is associated with compositional changes in intestinal microbiota. The level of glucose tolerance should be considered when linking microbiota with metabolic diseases such as obesity and developing strategies to control metabolic diseases by modifying the gut microbiota.
Disentangling type 2 diabetes and metformin treatment signatures in the human gut microbiota.
Forslund Kristoffer,Hildebrand Falk,Nielsen Trine,Falony Gwen,Le Chatelier Emmanuelle,Sunagawa Shinichi,Prifti Edi,Vieira-Silva Sara,Gudmundsdottir Valborg,Pedersen Helle K,Arumugam Manimozhiyan,Kristiansen Karsten,Voigt Anita Yvonne,Vestergaard Henrik,Hercog Rajna,Costea Paul Igor,Kultima Jens Roat,Li Junhua,Jørgensen Torben,Levenez Florence,Dore Joël, ,Nielsen H Bjørn,Brunak Søren,Raes Jeroen,Hansen Torben,Wang Jun,Ehrlich S Dusko,Bork Peer,Pedersen Oluf
In recent years, several associations between common chronic human disorders and altered gut microbiome composition and function have been reported. In most of these reports, treatment regimens were not controlled for and conclusions could thus be confounded by the effects of various drugs on the microbiota, which may obscure microbial causes, protective factors or diagnostically relevant signals. Our study addresses disease and drug signatures in the human gut microbiome of type 2 diabetes mellitus (T2D). Two previous quantitative gut metagenomics studies of T2D patients that were unstratified for treatment yielded divergent conclusions regarding its associated gut microbial dysbiosis. Here we show, using 784 available human gut metagenomes, how antidiabetic medication confounds these results, and analyse in detail the effects of the most widely used antidiabetic drug metformin. We provide support for microbial mediation of the therapeutic effects of metformin through short-chain fatty acid production, as well as for potential microbiota-mediated mechanisms behind known intestinal adverse effects in the form of a relative increase in abundance of Escherichia species. Controlling for metformin treatment, we report a unified signature of gut microbiome shifts in T2D with a depletion of butyrate-producing taxa. These in turn cause functional microbiome shifts, in part alleviated by metformin-induced changes. Overall, the present study emphasizes the need to disentangle gut microbiota signatures of specific human diseases from those of medication.
Agavins reverse the metabolic disorders in overweight mice through the increment of short chain fatty acids and hormones.
Huazano-García Alicia,López Mercedes G
Food & function
In this study, the effects of agavins (branched fructans) along with a diet shift on metabolic parameters, short chain fatty acid (SCFA) production and gastrointestinal hormones in overweight mice were established. Male C57BL/6 mice were fed with a standard (ST) or high fat (HF) diet over the course of 5 weeks, with the objective to induce overweightness in the animals, followed by a diet shift (HF_ST) and a diet shift with agavins (HF_ST + A) or inulin (HF_ST + O) for 5 additional weeks. After the first 5 weeks, the HF group showed a 30% body weight gain and an increase in glucose, triglyceride and cholesterol concentrations of 9%, 79% and 38% respectively when compared to the ST group (P < 0.05). Only the overweight mice that received agavins or inulin in their diets reversed the metabolic disorders induced by consumption of the HF diet, reaching the values very close to those of the ST group (P < 0.05). Furthermore, the consumption of agavins or inulin led to higher SCFA concentrations in the gut and modulated hormones such as GLP-1 and leptin involved in food intake regulation (P < 0.05). These findings demonstrate that a change of diet and fructan consumption such as agavins is a good alternative to increase weight loss and to improve the metabolic disorders associated with being overweight.
Microbiota-generated metabolites promote metabolic benefits via gut-brain neural circuits.
De Vadder Filipe,Kovatcheva-Datchary Petia,Goncalves Daisy,Vinera Jennifer,Zitoun Carine,Duchampt Adeline,Bäckhed Fredrik,Mithieux Gilles
Soluble dietary fibers promote metabolic benefits on body weight and glucose control, but underlying mechanisms are poorly understood. Recent evidence indicates that intestinal gluconeogenesis (IGN) has beneficial effects on glucose and energy homeostasis. Here, we show that the short-chain fatty acids (SCFAs) propionate and butyrate, which are generated by fermentation of soluble fiber by the gut microbiota, activate IGN via complementary mechanisms. Butyrate activates IGN gene expression through a cAMP-dependent mechanism, while propionate, itself a substrate of IGN, activates IGN gene expression via a gut-brain neural circuit involving the fatty acid receptor FFAR3. The metabolic benefits on body weight and glucose control induced by SCFAs or dietary fiber in normal mice are absent in mice deficient for IGN, despite similar modifications in gut microbiota composition. Thus, the regulation of IGN is necessary for the metabolic benefits associated with SCFAs and soluble fiber.
Dietary fructans, but not cellulose, decrease triglyceride accumulation in the liver of obese Zucker fa/fa rats.
Daubioul Catherine,Rousseau Nicolas,Demeure Roger,Gallez Bernard,Taper Henryk,Declerck Barbara,Delzenne Nathalie
The Journal of nutrition
This study was designed to compare the effects of dietary supplementation with nondigestible carbohydrates, differing in fermentability by colonic bacteria, on hepatic steatosis in growing obese Zucker rats. Male Zucker fa/fa rats were divided into three groups: a control group that received the basal diet, a fructan group that received 10 g highly fermented Synergy 1/100 g diet and a cellulose group that received 10 g poorly fermented Vivapur Microcrystalline cellulose/100 g diet. Rats consuming fructan had a lower energy intake, a lower body weight and less triacylglycerol accumulation in the liver as assessed in vivo by nuclear magnetic resonance (NMR) spectroscopy, and ex vivo by biochemical and histochemical analysis compared with the control and/or cellulose groups. The high fermentation of fructans compared with cellulose was reflected by greater cecal contents and by a twofold greater propionate concentration in the portal vein of rats fed fructan compared with those fed cellulose. By measuring the capacity of hepatocytes isolated from liver of Zucker rats to synthesize triglycerides or total lipids from different precursors, we showed that propionate, at the concentrations measured in the portal vein of rats treated with fructan, selectively decreased the incorporation of acetate into total lipids, a phenomenon that could contribute, along with the lower energy intake, to less triglyceride accumulation in the liver of obese Zucker rats fed dietary fructans.
The role of pH in determining the species composition of the human colonic microbiota.
Duncan Sylvia H,Louis Petra,Thomson John M,Flint Harry J
The pH of the colonic lumen varies with anatomical site and microbial fermentation of dietary residue. We have investigated the impact of mildly acidic pH, which occurs in the proximal colon, on the growth of different species of human colonic bacteria in pure culture and in the complete microbial community. Growth was determined for 33 representative human colonic bacteria at three initial pH values (approximately 5.5, 6.2 and 6.7) in anaerobic YCFA medium, which includes a mixture of short-chain fatty acids (SCFA) with 0.2% glucose as energy source. Representatives of all eight Bacteroides species tested grew poorly at pH 5.5, as did Escherichia coli, whereas 19 of the 23 gram-positive anaerobes tested gave growth rates at pH 5.5 that were at least 50% of those at pH 6.7. Growth inhibition of B. thetaiotaomicron at pH 5.5 was increased by the presence of the SCFA mix (33 mM acetate, 9 mM propionate and 1 mM each of iso-valerate, valerate and iso-butyrate). Analysis of amplified 16S rRNA sequences demonstrated a major pH-driven shift within a human faecal bacterial community in a continuous flow fermentor. Bacteroides spp. accounted for 27% of 16S rRNA sequences detected at pH 5.5, but 86% of sequences at pH 6.7. Conversely, butyrate-producing gram-positive bacteria related to Eubacterium rectale represented 50% of all 16S rRNA sequences at pH 5.5, but were not detected at pH 6.7. Inhibition of the growth of a major group of gram-negative bacteria at mildly acidic pH apparently creates niches that can be exploited by more low pH-tolerant microorganisms.
Starch utilization by the human large intestinal microflora.
Macfarlane G T,Englyst H N
The Journal of applied bacteriology
High levels (2-565 units/g) of amylase activity were observed in human faeces. Over 92% of amylase activity in faeces obtained from healthy persons was extracellular, whereas only about 9% of activity was associated with particulate material and washed cells. Bacterial cell-bound amylases were considerably more efficient in breaking down starch, however, than were the soluble enzymes which occurred in cell-free faecal supernatant fluids. Cell population densities of anaerobic starch-hydrolysing bacteria in the stools of ten persons ranged from 1.1 X 10(10) to 3.3 X 10(12)/g of faeces. Identification of 120 starch-hydrolysing colonies isolated from the stools of six subjects showed that the predominant amylolytic bacteria belonged to the genera Bifidobacterium, Bacteroides, Fusobacterium and Butyrivibrio. Mixed populations of gut bacteria rapidly fermented starch with the production of volatile fatty acids and organic acids. Lactate was observed to be a major, though transient intermediate during starch fermentation by these cultures. Approximately 60% of starch utilized was converted to volatile fatty acids, which in the human colon would be potentially available for absorption.
Nutraceutical potential of Aerva lanata (L.) Juss. ex Schult ameliorates secondary complications in streptozotocin-induced diabetic rats.
Riya M P,Antu K A,Pal S,Srivastava A K,Sharma S,Raghu K G
Food & function
Nutraceuticals provide health benefits beyond their basic nutrition by modulating a number of biochemical pathways. They are derived from natural products and have gained recognition worldwide as an adjuvant or therapy in the treatment of metabolic disorders such as diabetes. Although the regulation of blood glucose with drugs and insulin greatly reduces the incidence of secondary complications, the need for long-term treatment raises issues of tolerance and affordability. Therefore, the aim of the present study is to explore the nutraceutical potential of Aerva lanata, a herb widely used for its culinary and therapeutic potential in streptozotocin (STZ)-induced diabetic rats. Treatment with 70% ethanolic extract (ALE) at 500 mg per kg b.w per day for 21 days significantly improved the fasting blood glucose (120.33 ± 1.99 mg dL(-1)), insulin level (9.81 ± 0.38 mU L(-1)), HbA1c (7.3 ± 0.36%) and glycogen content in the liver (35.33 ± 1.38 mg g(-1) protein) and muscle (7.67 ± 0.11 mg g(-1) protein) compared to diabetic controls. The extract also showed a significant decrease in blood glucose by 47.29% towards the end of 2 h in oral glucose tolerance test on Day 21. Its therapeutic potential could be partly attributable to the presence of flavonoids, tannins and terpenes (alpha amyrin, betulin and beta sitosterol) along with micronutrients such as potassium, magnesium, calcium and zinc. Hence, we suggest the suitability of Aerva lanata as a nutraceutical for diabetic patients.
Contraction of gut smooth muscle cells assessed by fluorescence imaging.
Tokita Yohei,Akiho Hirotada,Nakamura Kazuhiko,Ihara Eikichi,Yamamoto Masahiro
Journal of pharmacological sciences
Here we discuss the development of a novel cell imaging system for the evaluation of smooth muscle cell (SMC) contraction. SMCs were isolated from the circular and longitudinal muscular layers of mouse small intestine by enzymatic digestion. SMCs were stimulated by test agents, thereafter fixed in acrolein. Actin in fixed SMCs was stained with phalloidin and cell length was determined by measuring diameter at the large end of phalloidin-stained strings within the cells. The contractile response was taken as the decrease in the average length of a population of stimulated-SMCs. Various mediators and chemically identified compounds of daikenchuto (DKT), pharmaceutical-grade traditional Japanese prokinetics, were examined. Verification of the integrity of SMC morphology by phalloidin and DAPI staining and semi-automatic measurement of cell length using an imaging analyzer was a reliable method by which to quantify the contractile response. Serotonin, substance P, prostaglandin E2 and histamine induced SMC contraction in concentration-dependent manner. Two components of DKT, hydroxy-α-sanshool and hydroxy-β-sanshool, induced contraction of SMCs. We established a novel cell imaging technique to evaluate SMC contractility. This method may facilitate investigation into SMC activity and its role in gastrointestinal motility, and may assist in the discovery of new prokinetic agents.
All-trans-configuration in Zanthoxylum alkylamides swaps the tingling with a numbing sensation and diminishes salivation.
Bader Matthias,Stark Timo D,Dawid Corinna,Lösch Sofie,Hofmann Thomas
Journal of agricultural and food chemistry
The methanol soluble prepared from a supercritical fluid extract of Szechuan pepper (Zanthoxylum piperitum) was screened for its key tingling and numbing chemosensates by application of taste dilution analysis. Further separation of fractions perceived with the highest sensory impact, followed by LC-TOF-MS, LC-MS, and 1D/2D NMR experiments, led to the structure determination of the known alkylamides hydroxy-γ-sanshool (1), hydroxy-α-sanshool (2), hydroxy-β-sanshool (3), bungeanool (4), isobungeanool (5), and hydroxy-γ-isosanshool (6), as well as hydroxy-ε-sanshool (7), the structure of which has not yet been confirmed by NMR, and hydroxy-ζ-sanshool (8), which has not been previously reported in the literature. Psychophysical half-tongue experiments using filter paper rectangles (1 × 2 cm) as the vehicle revealed amides 1, 2, 4, 5, 7, and 8, showing at least one cis-configured double bond, elicited the well-known tingling and paresthetic orosensation above threshold levels of 3.5-8.3 nmol/cm(2). In contrast, the all-trans-configured amides 3 and 6 induced a numbing and anesthetic sensation above thresholds of 3.9 and 7.1 nmol/cm(2), respectively. Interestingly, the mono-cis-configured major amide 2 was found to induce massive salivation, whereas the all-trans-configuration of 3 did not.
Pungent qualities of sanshool-related compounds evaluated by a sensory test and activation of rat TRPV1.
Sugai Etsuko,Morimitsu Yasujiro,Iwasaki Yusaku,Morita Akihito,Watanabe Tatsuo,Kubota Kikue
Bioscience, biotechnology, and biochemistry
The detection threshold and taste characteristics of sanshools were examined by sensory evaluation, after isolating four sanshools (alpha-, beta-, gamma-, and delta-), and two hydroxy sanshools (alpha- and beta-) from the pericarp of Japanese pepper. The Scoville unit (SU) values of the four sanshools were in the range of 80,000-110,000, while those of hydroxy sanshools were 3-5 fold lower than corresponding sanshools. The pungent qualities of each sanshool were different. Burning and tingling were predominantly perceived and lasted for the longest time with alpha-sanshool. Burning and fresh for gamma-sanshool, and tingling and numbing for hydroxy alpha-sanshool were perceived. Tests on the activation of rat TRPV1 were also performed. All of them were weak agonists. Among them, gamma-sanshool was the most potent agonist, although its EC50 value of 5.3 microM was 230 fold higher than that of capsaicin. These results indicate that it would be difficult to explain the pungent quality of each sanshool simply in terms of TRPV1 activation.
Gut microorganisms as promising targets for the management of type 2 diabetes.
Delzenne Nathalie M,Cani Patrice D,Everard Amandine,Neyrinck Audrey M,Bindels Laure B
Each human intestine harbours not only hundreds of trillions of bacteria but also bacteriophage particles, viruses, fungi and archaea, which constitute a complex and dynamic ecosystem referred to as the gut microbiota. An increasing number of data obtained during the last 10 years have indicated changes in gut bacterial composition or function in type 2 diabetic patients. Analysis of this 'dysbiosis' enables the detection of alterations in specific bacteria, clusters of bacteria or bacterial functions associated with the occurrence or evolution of type 2 diabetes; these bacteria are predominantly involved in the control of inflammation and energy homeostasis. Our review focuses on two key questions: does gut dysbiosis truly play a role in the occurrence of type 2 diabetes, and will recent discoveries linking the gut microbiota to host health be helpful for the development of novel therapeutic approaches for type 2 diabetes? Here we review how pharmacological, surgical and nutritional interventions for type 2 diabetic patients may impact the gut microbiota. Experimental studies in animals are identifying which bacterial metabolites and components act on host immune homeostasis and glucose metabolism, primarily by targeting intestinal cells involved in endocrine and gut barrier functions. We discuss novel approaches (e.g. probiotics, prebiotics and faecal transfer) and the need for research and adequate intervention studies to evaluate the feasibility and relevance of these new therapies for the management of type 2 diabetes.
Roles of stem cell factor on the depletion of interstitial cells of Cajal in the colon of diabetic mice.
Lin Lin,Xu Li-ming,Zhang Wei,Ge Ying-bin,Tang Yu-rong,Zhang Hong-jie,Li Xue-liang,Chen Jiande D Z
American journal of physiology. Gastrointestinal and liver physiology
The aim of this study was to investigate the effects of stem cell factor (SCF) on interstitial cell of Cajal (ICC) depletion in the colon of diabetic mice. Male C57/BL6 mice were treated by a single intraperitoneally injected dose of streptozotocin, and those displaying sustained high blood glucose were selected as diabetes mellitus models. Six groups of mice were used: three groups of normal nondiabetic mice (untreated and treated with IgG or SCF antibody), and three groups of diabetic mice (untreated and treated with vehicle or SCF). Changes of the ICC quantities were analyzed by immunohistochemistry. ICC morphologies were observed with transmission electron microscopy. The SCF levels in sera and colon tissues were detected by ELISA and Western blot, respectively. The nondiabetic mice treated with SCF antibody and the untreated diabetic mice showed decreased SCF levels in the sera and colonic tissues, reduced numbers of ICC, and pathological changes of the ICC ultrastructures, whereas the nondiabetic mice treated with mouse IgG showed no significant changes compared with the nondiabetic mice. The diabetic mice treated with exogenous SCF showed restored SCF levels in both sera and colon tissues and improvement in the numbers of ICC and the damages of ICC ultrastructures, whereas the vehicle control of diabetic mice showed no significant changes compared with the diabetic mice. The blood glucose remained high and unchanged with the treatment of SCF or vehicle in the diabetic mice. These results indicate that diabetic mice show a decline in the number of ICC and impairment in the ultrastructures of ICC, and these abnormalities are attributed to a deficiency in the endogenous SCF but are not related to hyperglycemia. Exogenous SCF partially reverses the pathological changes of ICC in diabetic mice.
Effect of Kaiyu Qingwei Jianji on the morphometry and residual strain distribution of small intestine in experimental diabetic rats.
Sha Hong,Zhao Jing-Bo,Zhang Zhi-Yuan,Zhou Shui-Ping,Tong Xiao-Lin,Zhuang Feng-Yuan,Gregersen Hans
World journal of gastroenterology
AIM:To investigate the effect of a Chinese medicine, Kaiyu Qingwei Jianji (KYQWJJ) used for diabetic treatment, on the morphometry and residual strain distribution of the small intestine in streptozotocin (STZ) -induced diabetic rats. Correlation analysis was also performed between the opening angle and residual strain with the blood glucose level. METHODS:Forty-two male Wistar rats weighing 220-240 g were included in this study. Thirty-two STZ-induced diabetic rats were subdivided into four groups (n = 8 in each group), i.e. diabetic control group (DM); high dose of KYQWJJ (T1, 36 g/kg per day); low dose of KYQWJJ (T2, 17 g/kg per day) and Gliclazide (T3, 50 mg/kg per day). Another ten rats were used as non-diabetic control (CON). The medicines were poured directly into stomach lumen by gastric lavage twice daily. The rats of CON and DM groups were only poured the physiological saline. Blood glucose and plasma insulin levels were measured. Experimental period was 35 d. At the end of experiment, three 5-cm long segments were harvested from the duodenum, jejunum and ileum. Three rings of 1-2 mm in length for no-load and zero-stress state tests were cut from the middle of different segments. The morphometric data, such as the circumferential length, the wall thickness and the opening angle were measured from the digitized images of intestinal segments in the no-load state and zero-stress state. The residual strain was computed from the morphometry data. Furthermore, the linear regression analysis was performed between blood glucose level with morphometric and biomechanical data in the different intestinal segments. RESULTS:The blood glucose level of DM group was consistent 4-fold to 5-fold higher than those in CON group during the experiment (16.89+/-1.11 vs 3.44+/-0.15 mmol/L, P < 0.001). The blood glucose level in the T1 (16.89+/-1.11 vs 11.08+/-2.67 mmol/L, P < 0.01) and T3 groups (16.89+/-1.11 vs 13.54+/-1.73 mmol/L, P < 0.05), but not in T2 group (P > 0.05) was significantly lower than those in DM group. The plasma insulin levels of DM, T1, T2 and T3 groups were significantly lower than those in CON group (10.98+/-1.02, 12.52+/-1.42,13.54+/-1.56,10.96+/-0.96 vs 17.84+/-2.34 pmol/L respectively, P < 0.05), but no significantly difference among the groups with exception of CON group. The wet weight/cm and total wall thickness of duodenum, jejunum and ileum in DM group were significantly higher than those in CON group (wet weight (g/cm): duodenum 0.209+/-0.012 vs 0.166+/-0.010, jejunum 0.149+/-0.008 vs 0.121+/-0.004, ileum 0.134+/-0.013 vs 0.112+/-0.007; Wall thickness (mm): duodenum 0.849+/-0.027 vs 0.710+/-0.026, jejunum 0.7259+/-0.034 vs 0.627+/-0.025, ileum 0.532+/-0.023 vs 0.470+/-0.010, all P < 0.05), T1 and T3 treatment could partly restore change of wall thickness, but T2 could not. The opening angle and absolute value of inner and outer residual stain were significantly smaller in duodenal segment (188+/-11 degrees, -0.31+/-0.02 and 0.35+/-0.03 vs 259+/-15 degrees, -0.40+/-0.02 and 0.43+/-0.05) and larger in jejunal (215+/-20 degrees, -0.30+/-0.03 and 0.36+/-0.06 vs 172+/-19 degrees, -0.25+/-0.02 and 0.27+/-0.02) and ileal segments (183+/-20 degrees, -0.28+/-0.01 and 0.34+/-0.05 vs 153+/-14 degrees, -0.23+/-0.03 and 0.29+/-0.04) in DM group than in CON group (P < 0.01). T1 and T3 treatment could partly restore this biomechanical alteration, but strong effect was found in T1 treatment (duodenum 243+/-14 degrees, -0.36+/-0.02 and 0.42+/-0.06, jejunum 180+/-15 degrees, -0.26+/-0.03 and 0.30+/-0.06 and ileum 163+/-17 degrees, -0.23+/-0.03 and 0.30+/-0.05, compared with DM, P < 0.05). The linear association was found between the glucose level with most morphometric and biomechanical data. CONCLUSION:KYQWJJ (high dose) treatment could partly restore the changes of blood glucose level and the remodeling of morphometry and residual strain of small intestine in diabetic rats. The linear regression analysis demonstrated that the effect of KYQWJJ on intestinal opening angle and residual strain is partially through its effect on the blood glucose level.
Role of the intestinal tight junction modulator zonulin in the pathogenesis of type I diabetes in BB diabetic-prone rats.
Watts Tammara,Berti Irene,Sapone Anna,Gerarduzzi Tania,Not Tarcisio,Zielke Ronald,Fasano Alessio
Proceedings of the National Academy of Sciences of the United States of America
Increased intestinal permeability has been observed in numerous human autoimmune diseases, including type-1 diabetes (T1D) and its' animal model, the BB-wor diabetic prone rat. We have recently described zonulin, a protein that regulates intercellular tight junctions. The objective of this study was to establish whether zonulin-dependent increased intestinal permeability plays a role in the pathogenesis of T1D. In the BB diabetic-prone rat model of T1D, intestinal intraluminal zonulin levels were elevated 35-fold compared to control BB diabetic-resistant rats. Zonulin up-regulation was coincident with decreased small intestinal transepithelial electrical resistance, and was followed by the production of autoantibodies against pancreatic beta cells, which preceded the onset of clinically evident T1D by approximately 25 days. In those diabetic prone rats that did not progress to diabetes, both intraluminal zonulin and transepithelial electrical resistance were similar to those detected in diabetic-resistant animal controls. Blockade of the zonulin receptor reduced the cumulative incidence of T1D by 70%, despite the persistence of intraluminal zonulin up-regulation. Moreover, treatment responders did not seroconvert to islet cell antibodies. Combined together, these findings suggest that the zonulin-induced loss in small intestinal barrier function is involved in the pathogenesis of T1D in the BB diabetic-prone animal model.
Cloning and molecular characterization of the ontogeny of a rat ileal sodium-dependent bile acid transporter.
Shneider B L,Dawson P A,Christie D M,Hardikar W,Wong M H,Suchy F J
The Journal of clinical investigation
Sodium-dependent bile acid transport in the rat ileum is abruptly expressed at weaning. Degenerate oligonucleotides, based on amino acid sequence identities between the rat liver and hamster ileal transporters, were used to amplify a rat ileal probe. A 1.2-kb cDNA clone, which contains the full coding region (348 amino acids, 38 kD), was isolated by hybridization screening. In vitro translation yielded a 38-kD protein which glycosylated to 48 kD. Sodium-dependent uptake of taurocholate was observed in oocytes injected with cRNA. Northern blot analysis revealed a 5.0-kb mRNA in ileum, kidney, and cecum. A 48-kD protein was detected in ileal brush border membranes and localized to the apical border of villus ileal enterocytes. mRNA and protein expression, which were negligible before weaning, increased dramatically at weaning. Nuclear transcription rates for the transporter increased 15-fold between postnatal days 7 and 28. The apparent molecular weight of the transporter also increased between days 19 and 28. In summary, the developmental regulation of the rat ileal sodium-dependent bile acid cotransporter is characterized by transcriptionally regulated increases in mRNA and protein levels at the time of weaning with changes in apparent molecular weight of the protein after weaning.
Nonfasting triglycerides and risk of myocardial infarction, ischemic heart disease, and death in men and women.
Nordestgaard Børge G,Benn Marianne,Schnohr Peter,Tybjaerg-Hansen Anne
CONTEXT:Elevated nonfasting triglycerides indicate the presence of remnant lipoproteins, which may promote atherosclerosis. OBJECTIVE:To test the hypothesis that very high levels of nonfasting triglycerides predict myocardial infarction (MI), ischemic heart disease (IHD), and death. DESIGN, SETTING, AND PARTICIPANTS:A prospective cohort study of 7587 women and 6394 men from the general population of Copenhagen, Denmark, aged 20 to 93 years, followed up from baseline (1976-1978) until 2004. MAIN OUTCOME MEASURES:Hazard ratios (HRs) for incident MI, IHD, and total death according to baseline nonfasting triglyceride level categories of 1 to 1.99 mmol/L (88.5-176.1 mg/dL), 2 to 2.99 mmol/L (177.0-264.6 mg/dL), 3 to 3.99 mmol/L (265.5-353.0 mg/dL), 4 to 4.99 mmol/L (354.0-441.6 mg/dL), and 5 mmol/L or more (> or =442.5 mg/dL) vs triglyceride levels of less than 1 mmol/L (<88.5 mg/dL). RESULTS:With increasing levels of nonfasting triglycerides, levels of remnant lipoprotein cholesterol increased. During a mean follow-up of 26 years, 1793 participants (691 women and 1102 men) developed MI, 3479 (1567 women and 1912 men) developed IHD, and 7818 (3731 women and 4087 men) died. For MI, among women, the age-adjusted HRs and multifactorially adjusted HRs (aHRs) for each respective category per 1-mmol/L increase in nonfasting triglyceride levels were 2.2 (aHR, 1.7), 4.4 (aHR, 2.5), 3.9 (aHR, 2.1), 5.1 (aHR, 2.4), and 16.8 (aHR, 5.4); for both, P for trend < .001. For MI, among men, the values were 1.6 (aHR, 1.4), 2.3 (aHR, 1.6), 3.6 (aHR, 2.3), 3.3 (aHR, 1.9), and 4.6 (aHR, 2.4); for both, P for trend < .001. For IHD, among women, the values were 1.7 (aHR, 1.4), 2.8 (aHR, 1.8), 3.0 (aHR, 1.8), 2.1 (aHR, 1.2), and 5.9 (aHR, 2.6); for both, P for trend < .001. For IHD, among men, the values were 1.3 (aHR, 1.1), 1.7 (aHR, 1.3), 2.1 (aHR, 1.3), 2.0 (aHR, 1.2), and 2.9 (aHR, 1.5); P for trend < .001 for age-adjusted and P for trend = .03 for multifactorially adjusted. For total death, among women, the values were 1.3 (aHR, 1.3), 1.7 (aHR, 1.6), 2.2 (aHR, 2.2), 2.2 (aHR, 1.9), and 4.3 (aHR, 3.3); for both, P for trend < .001. For total death, among men, the values were 1.3 (aHR, 1.2), 1.4 (aHR, 1.4), 1.7 (aHR, 1.5), 1.8 (aHR, 1.6), and 2.0 (aHR, 1.8); for both, P for trend < .001. CONCLUSION:In this general population cohort, elevated nonfasting triglyceride levels were associated with increased risk of MI, IHD, and death in men and women.
The effect of high-amylose cornstarch on lipid metabolism in OVX rats is affected by fructose feeding.
Liu Xiong,Ogawa Hiroshi,Kishida Taro,Ebihara Kiyoshi
The Journal of nutritional biochemistry
We examined whether the effects of high-amylose cornstarch (HACS) on lipid metabolism in ovariectomized (OVX) rats were affected by high-fructose feeding. Sucrose (482 g/kg diet) was used as fructose source. OVX rats were fed one of the following four diets for 21 days: a sucrose-based or cornstarch-based cholesterol-free diet with or without HACS (150 g/kg diet). Body weight and food intake were increased by sucrose. Plasma total cholesterol and low-density lipoprotein cholesterol concentrations were increased by sucrose and decreased by HACS in cornstarch-fed rats, but not in sucrose-fed rats. Liver total lipids and concentrations of plasma and liver triacylglycerol (TAG) were increased by sucrose, whereas plasma TAG concentration was decreased by HACS, in sucrose-fed rats. However, liver cholesterol concentration was not affected by diet. The amount of cholesterol in small-intestinal contents was increased in sucrose-fed rats, but not in cornstarch-fed rats, but that of bile acids was not affected by diet. Fecal excretions of bile acids and neutral sterols were increased by HACS. The level of sterol-regulatory element-binding protein-1c mRNA was increased by sucrose and decreased by HACS in sucrose-fed rats, but not in cornstarch-fed rats. The level of farnesoid X receptor mRNA was decreased by sucrose and increased by HACS in cornstarch-fed rats, but not in sucrose-fed rats, as was the level of cholesterol 7alpha-hydroxylase mRNA. These results show that the effect of HACS on hyperlipidemia induced by ovarian hormone deficiency would be affected by the consumption of fructose-rich sweeteners such as sucrose and high-fructose syrup.
Effect of dietary resistant starch and protein on colonic fermentation and intestinal tumourigenesis in rats.
Le Leu Richard K,Brown Ian L,Hu Ying,Morita Tatsuya,Esterman Adrian,Young Graeme P
Protein as well as starch is fermented in the colon, but the interaction between protein and starch fermentation and the impact on colonic oncogenesis is unknown. High-protein diets increase delivery of protein to the colon and might promote oncogenesis through generation of toxic products. We investigated the interaction of resistant starch (RS) with digestion-resistant potato protein (PP) on colonic fermentation events and their relationship to intestinal tumourigenesis. Male Sprague-Dawley rats were fed an AIN-76A-based diet for 4 weeks and intestinal neoplasms were induced by azoxymethane. Experimental diets included the following: no added RS or PP, 10% high amylose maize starch (source of RS) replacing digestible starch, 15% PP replacing casein and 10% high amylose maize starch+15% PP. Rats were maintained on diets until killed at 30 weeks. Feeding RS significantly increased short-chain fatty acid (SCFA) levels (P<0.001) in the caecum and colon. Importantly, butyrate concentration was significantly increased in the distal colon with RS (P<0.001). Feeding PP increased protein fermentation products, but this effect was reduced by adding RS to the diet. Intestinal neoplasms and colorectal adenocarcinomas were reduced by feeding RS (P<0.01) regardless of whether PP was fed, whereas PP alone increased the incidence and number of small intestinal neoplasms including the adenocarcinomas (P<0.01). In conclusion, RS altered the colonic luminal environment by increasing the concentration of SCFAs including butyrate and lowering production of potentially toxic protein fermentation products. These effects of RS not only protected against intestinal tumourigenesis but also ameliorated the tumour-enhancing effects of feeding indigestible protein.
Chemical constituents and biological activities of the fruit of Zanthoxylum integrifoliolum.
Chen I S,Chen T L,Chang Y L,Teng C M,Lin W Y
Journal of natural products
Through continuing studies on the chemical constituents and antiplatelet aggregation principles of the fruit of the Formosan Zanthoxylum integrifoliolum, four new compounds-including two new lignans, (+)-pinoresinol-di-3,3-dimethylallyl ether (1), and (+)-pinoresinol-3,3-dimethylallyl ether (2); zanthonitrile (3), and one new flavonoid, 3,5-diacetyltambulin (4)-and 18 known compounds were isolated from the CHCl3-soluble fraction. Their structures were elucidated on the basis of spectral data and chemical evidence. Among the isolates, including the previously reported isobutylamides, 13 compounds showed strong in vitro antiplatelet aggregation activity, with only (-)-tetrahydroberberine showing weak vasorelaxing effect in high potassium- or norepinephrine-induced contraction of rat aorta.
Inhibition of liver microsomal cytochrome P450 activity and metabolism of the tobacco-specific nitrosamine NNK by capsaicin and ellagic acid.
Zhang Z,Hamilton S M,Stewart C,Strother A,Teel R W
The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3- pyridyl)-1-butanone (NNK), present in tobacco and tobacco smoke, is metabolically activated by microsomal enzymes. In this study, we examined the effect of capsaicin and ellagic acid on the in vitro metabolism of NNK by hamster and rat liver microsomes. Capsaicin is the principal component of Capsicum fruits used widely throughout the world as a food additive. Ellagic acid, with reported anticarcinogenic properties, is found in various soft fruits and nuts. Both capsaicin and ellagic acid inhibited the major pathways of NNK-reduction, N-pyridine oxidation and a-hydroxylation by hamster liver microsomes. Capsaicin inhibited NNK-reduction and a-hydroxylation and ellagic acid inhibited N-oxidation and a-hydroxylation by rat liver microsomes. The effects of capsaicin and ellagic acid on isozymes of cytochrome P450 were observed in the hydroxylation reactions of the metabolism of the steroid hormone testosterone. Results of these experiments indicated that both capsaicin and ellagic acid strongly inhibited the constitutive enzymes CYP 2A2, 3A1, 2C11, 2B1, 2B2 and 2C6. This study suggests that capsaicin and ellagic acid, as naturally occurring dietary constituents, possess antimutagenic and anticarcinogenic properties through the inhibition of xenobiotic metabolizing enzymes.
Beneficial influence of dietary curcumin, capsaicin and garlic on erythrocyte integrity in high-fat fed rats.
Kempaiah Rayavara K,Srinivasan Krishnapura
The Journal of nutritional biochemistry
In rats rendered hyperlipidemic by maintaining them on a high-fat diet (30%) for 8 weeks, inclusion of spice principles [curcumin (0.2%) or capsaicin (0.015%)] or garlic (2.0%) in the diet produced significant hypotriglyceridemic effect. Plasma cholesterol remained unaffected in high-fat treatment. Hepatic triglyceride content was significantly higher in high-fat fed rats, and this increase was effectively countered by inclusion of the hypolipidemic spice agents -- curcumin, capsaicin or garlic in the diet. The lipid profile of erythrocyte membranes of hyperlipidemic rats was similar to basal controls. An examination of the osmotic fragility of erythrocytes in various groups indicated that the red blood cells of hyperlipidemic rats display a slight resistance to osmotic lysis. Inclusion of spice principles [curcumin (0.2%) or capsaicin (0.015%)] or garlic (2.0%) in the diet, which produced the hypotriglyceridemic effect, appeared to beneficially correct this altered osmotic fragility of erythrocytes. Activities of ouabain-sensitive Na(+),K(+)-ATPase as well as acetylcholinesterase of erythrocyte membranes in high-fat fed rats remained unaltered. Activity of Ca(2+),Mg(2+)-ATPase in erythrocyte membrane was significantly decreased in high-fat fed animals, whereas dietary spice principles and garlic countered this reduction in enzyme activity. In the absence of any change in the cholesterol/phospholipid molar ratio in the erythrocyte membrane, a decreased activity of membrane-bound Ca(2+),Mg(2+)-ATPase could have probably contributed to the accumulation of intracellular calcium leading to the diminished deformability of the erythrocytes in high-fat fed rats.
Integrity of erythrocytes of hypercholesterolemic rats during spices treatment.
Kempaiah R K,Srinivasan K
Molecular and cellular biochemistry
In rats rendered hypercholesterolemic by maintaining them on a cholesterol-enriched diet (0.5%) for 8 weeks, inclusion of spice principles--curcumin (0.2%) or capsaicin (0.015%) or the spice--garlic powder (2.0%) in the diet, produced the expected hypolipidemic effect. Plasma cholesterol which was more than 200% that of basal control in hypercholesterolemic rats, was decreased by these dietary spice principles and garlic by 25-39%. Erythrocyte membranes of hypercholesterolemic rats were relatively enriched in cholesterol, which was about 120% of basal control, while membrane phospholipid was unaffected. This resulted in a significant alteration in cholesterol to phospholipid ratio of RBC membranes. Dietary curcumin, capsaicin and garlic were observed to counter this altered lipid profile of erythrocyte membranes in hypercholesterolemic situation by producing a significant 10-14% decrease in membrane cholesterol content. As a result of alteration in membrane structural lipids, the structural integrity of RBCs was also affected. An examination of the osmotic fragility of erythrocytes in various groups, indicated that RBCs of hypercholesterolemic rats were relatively fragile compared to normal controls. Dietary curcumin, capsaicin and garlic appeared to correct this increased fragility of erythrocytes.
Effect of capsaicin on substrate oxidation and weight maintenance after modest body-weight loss in human subjects.
Lejeune Manuela P G M,Kovacs Eva M R,Westerterp-Plantenga Margriet S
The British journal of nutrition
The aim of the present study was to investigate whether capsaicin assists weight maintenance by limiting weight regain after weight loss of 5 to 10 %. In this randomized double-blind placebo-controlled study, ninety-one moderately overweight subjects were randomly assigned to an intensive group that underwent all the measurements, and an extensive group that underwent the same measurements except the metabolism measurements. After a 4-week very-low-energy diet (VLED) intervention, a 3-month weight-maintenance period followed. During weight maintenance, subjects were divided into a capsaicin (135 mg capsaicin/d) and a placebo group. Body mass was measured before and after the VLED and after 1, 2 and 3 months of weight maintenance. The mean body-mass loss during the VLED was 6.6 (SD 2.0) kg (7.8 (SD 1.8) % initial body mass), and was not different between the subsequent treatment and placebo group. During weight maintenance, mean % regain during treatment was not significantly different compared with placebo (33.3 (SD 35.7) v. 19.2 (SD 41.8) %, P=0.09). RQ was significantly less increased during weight maintenance in the treatment group compared with placebo (0.04 (SD 0.06) v. 0.07 (SD 0.05), P<0.05), indicating a relatively more sustained fat oxidation. Fat oxidation (g/h) after weight maintenance was higher in the capsaicin group compared with placebo (4.2 (SD 1.1) v. 3.5 (SD 0.9), P<0.05). These results indicate that capsaicin treatment caused sustained fat oxidation during weight maintenance compared with placebo. However, capsaicin treatment has no limiting effect on 3-month weight regain after modest weight loss.
Effects of novel capsinoid treatment on fatness and energy metabolism in humans: possible pharmacogenetic implications.
The American journal of clinical nutrition
BACKGROUND:Capsinoids from the Capsicum genus of plants are nonpungent capsaicin-related substances with effects on metabolism and body weight in animals. OBJECTIVES:Our objectives were to explore the safety and efficacy of capsinoids taken orally (6 mg/d) for weight loss, fat loss, and change in metabolism and to examine whether candidate genes are predictors of capsinoid response. DESIGN:This was a 12-wk, placebo-controlled, double-blind, randomized study. Eligibility criteria included a body mass index (BMI; in kg/m(2)) of 25-35. Body weight was measured, and dual-energy X-ray absorptiometry, indirect calorimetry (men only), and genotyping were conducted. RESULTS:Forty women and 40 men with a mean (+/- SD) age of 42 +/- 8 y and BMI of 30.4 +/- 2.4 were randomly assigned to a capsinoid or placebo group. Capsinoids were well tolerated. Mean (+/- SD) weight change was 0.9 +/- 3.1 and 0.5 +/- 2.4 kg in the capsinoid and placebo groups, respectively (P = 0.86). There was no significant group difference in total change in adiposity, but abdominal adiposity decreased more (P = 0.049) in the capsinoid group (-1.11 +/- 1.83%) than in the placebo group (-0.18 +/- 1.94%), and this change correlated with the change in body weight (r = 0.46, P < 0.0001). Changes in resting energy expenditure did not differ significantly between groups, but fat oxidation was higher at the end of the study in the capsinoid group (least-squares mean difference: 21.0 mg/min; P = 0.06). Of 13 genetic variants tested, TRPV1 Val585Ile and UCP2 -866 G/A correlated significantly with change in abdominal adiposity. CONCLUSIONS:Treatment with 6 mg/d capsinoids orally appeared to be safe and was associated with abdominal fat loss. Capsinoid ingestion was associated with an increase in fat oxidation that was nearly significant. We identified 2 common genetic variants that may be predictors of therapeutic response.
Antinociceptive and anti-inflammatory activities of ethyl acetate fraction from Zanthoxylum armatum in mice.
Guo Tao,Deng Yun-Xia,Xie Hui,Yao Chun-Yan,Cai Cheng-Cheng,Pan Sheng-li,Wang Yang-Lin
Zanthoxylum armatum DC. is a traditional Chinese medicine that is prescribed to alleviate pain and treat inflammatory disorders. This species is distributed mainly in the southeast and southwest regions of China. In the present study, we found that ethyl acetate fraction of ethanolic extract of Z. armatum could significantly decrease acetic acid-induced writhing numbers, and suppress formalin induced licking times in the first phase at the highest dose and in the second phase at all tested doses. This observation revealed that Z. armatum extract possessed powerful antinociceptive activity. The mechanisms of the antinociceptive effect might be mainly involved in the periphery inflammatory analgesic. In addition, the ethyl acetate fraction also inhibited xylene-induced ear swelling in a dose-dependent manner in mice. Eight lignans [eudesmin, horsfieldin, fargesin, kobusin, sesamin, asarinin, planispine A, and pinoresinol-di-3,3-dimethylallyl] were identified as major components of the ethyl acetate fraction. Considering related studies reporting the anti-inflammatory activity for the identified lignans, lignan might be responsible for its anti-inflammatory activity. Our results confirm that the traditional use of Z. armatum in the treatment of inflammation and pain is warranted.
The effect of enzyme supplementation on the apparent metabolizable energy and nutrient digestibilities of wheat, barley, oats, and rye for the young broiler chick.
Friesen O D,Guenter W,Marquardt R R,Rotter B A
The influence of enzyme supplementation on the bioavailable energy (AME(n)) and apparent digestibilities of lipid (ALD) and protein (APD) in young broiler chicks was examined for diets containing either wheat, hulled or hulless barley, naked oats, or spring rye. Dietary AME(n), APD, and ALD values were depressed (P less than or equal to .01) for all test grains (except hulled Bedford barley) as the inclusion rate of the grain replacing wheat increased. The antinutritives, beta-glucans (barley and oats) and pentosans (rye), had the most pronounced effect on ALD. The decreases in ALD were 43, 77, and 67% for chicks fed diets containing 70% Scout barley (hulless), Terra oats, and Gazelle rye, respectively, compared with those fed the control wheat diet. Enzyme supplementation increased (P less than or equal to .01) AME(n), APD, and ALD for all test cereals. The corresponding increases in the AME(n), of the enzyme-supplemented diets containing 70% HY320 wheat, Bedford barley, Scout barley, Terra oats, and Gazelle rye diets were 4, 7, 42, 33, and 14%, respectively, compared with their unsupplemented counterparts. Enzyme treatment also improved (P less than or equal to .01) weight gains and feed conversion efficiencies of chicks fed diets containing each of the cereals. Overall, the results demonstrate that the nutritive value of cereal grains such as wheat, barley, oats, and rye can be improved by the addition of crude fungal extracts to the diet of young chicks.
Mitochondria in cancer: at the crossroads of life and death.
Fogg Vanessa C,Lanning Nathan J,Mackeigan Jeffrey P
Chinese journal of cancer
Mitochondrial processes play an important role in tumor initiation and progression. In this review, we focus on three critical processes by which mitochondrial function may contribute to cancer: through alterations in glucose metabolism, the production of reactive oxygen species (ROS) and compromise of intrinsic apoptotic function. Alterations in cancer glucose metabolism include the Warburg effect, leading to a shift in metabolism away from aerobic respiration toward glycolysis, even when sufficient oxygen is present to support respiration. Such alterations in cellular metabolism may favor tumor cell growth by increasing the availability of biosynthetic intermediates needed for cellular growth and proliferation. Mutations in specific metabolic enzymes, namely succinate dehydrogenase, fumarate hydratase and the isocitrate dehydrogenases, have been linked to human cancer. Mitochondrial ROS may contribute to cancer via DNA damage and the activation of aberrant signaling pathways. ROS-dependent stabilization of the transcription factor hypoxia-inducible factor (HIF) may be a particularly important event for tumorigenesis. Compromised function of intrinsic apoptosis removes an important cellular safeguard against cancer and has been implicated in tumorigenesis, tumor metastasis, and chemoresistance. Each of the major mitochondrial processes is linked. In this review, we outline the connections between them and address ways these mitochondrial pathways may be targeted for cancer therapy.
Expression of apical membrane L-glutamate transporters in neonatal porcine epithelial cells along the small intestinal crypt-villus axis.
Fan Ming Z,Matthews James C,Etienne Nadege M P,Stoll Barbara,Lackeyram Dale,Burrin Douglas G
American journal of physiology. Gastrointestinal and liver physiology
Enteral l-glutamate is extensively utilized as an oxidative fuel by the gut mucosa in the neonate. To identify major uptake pathways and to understand uptake regulation, we examined transport kinetics and molecular identities of apical membrane l-glutamate transporters in epithelial cells sequentially isolated along the small intestinal crypt-villus axis from milk protein-fed, 16-day-old pigs. The distended intestinal sac method was used to isolate 12 sequential cell fractions from the tip villus to the bottom crypt. Initial rates and kinetics of l-glutamate uptake were measured with l-[G-(3)H]glutamate by fast filtration in apical membrane vesicles prepared by Mg(2+) precipitation and differential centrifugation, with membrane potential clamped by SCN(-). Initial l-glutamate uptake results suggested the presence of B(o) and X(AG)(-) transport systems, but the X(AG)(-) system was predominant for uptake across the apical membrane. Kinetic data suggested that l-glutamate uptake through the X(AG)(-) system was associated with higher maximal transport activity but lower transporter affinity in crypt than in villus cells. Molecular identity of the X(AG)(-) glutamate transporter, based on immunoblot and RT-PCR analysis, was primarily the defined excitatory amino acid carrier (EAAC)-1. EAAC-1 expression was increased with cell differentiation and regulated at transcription and translation levels from crypt to upper villus cells. In conclusion, efficiency and capacity of luminal l-glutamate uptake across the apical membrane are regulated by changing expression of the X(AG)(-) system transporter gene EAAC-1 at transcription and translation levels as well as maximal uptake activity and transporter affinity along the intestinal crypt-villus axis in the neonate.
The aromatic amino acid tryptophan stimulates skeletal muscle IGF1/p70s6k/mTor signaling in vivo and the expression of myogenic genes in vitro.
Dukes Amy,Davis Colleen,El Refaey Mona,Upadhyay Sunil,Mork Sarah,Arounleut Phonepasong,Johnson Maribeth H,Hill William D,Isales Carlos M,Hamrick Mark W
Nutrition (Burbank, Los Angeles County, Calif.)
OBJECTIVES:Nutrition plays a key role in the maintenance of muscle and bone mass, and dietary protein deficiency has in particular been associated with catabolism of both muscle and bone tissue. One mechanism thought to link protein deficiency with loss of muscle mass is deficiency in specific amino acids that play a role in muscle metabolism. The aim of this study was to test the hypothesis that the essential amino acid tryptophan, and its metabolite kynurenine, might directly affect muscle metabolism in the setting of protein deficiency. METHODS:Adult mice (12 mo) were fed a normal diet (18% protein), as well as diets with low protein (8%) supplemented with increasing concentrations (50, 100, and 200 uM) of kynurenine (Kyn) or with tryptophan (Trp; 1.5 mM) for 8 weeks. Myoprogenitor cells were also treated with Trp and Kyn in vitro to determine their effects on cell proliferation and expression of myogenic differentiation markers. RESULTS:All mice on the low-protein diets weighed less than the group fed normal protein (18%). Lean mass measured by dual-energy X-ray absorptiometry was lowest in mice on the high Kyn diet, whereas percent lean mass was highest in mice receiving Trp supplementation and percent body fat was lowest in mice receiving Trp. Enzyme-linked immunosorbent assays showed significant increases in skeletal muscle insulin-like growth factor-1, leptin, and the myostatin antagonist follistatin with Trp supplementation. mRNA microarray and gene pathway analysis performed on muscle samples demonstrate that mTor/eif4/p70s6k pathway molecules are significantly up-regulated in muscles from mice on Kyn and Trp supplementation. In vitro, neither amino acid affected proliferation of myoprogenitors, but Trp increased the expression of the myogenic markers MyoD, myogenin, and myosin heavy chain. CONCLUSION:These findings suggest that dietary amino acids can directly affect molecular signaling in skeletal muscle, further indicating that dietary manipulation with specific amino acids could potentially attenuate muscle loss with dietary protein deficiency.
Identification of CB1/CB2 ligands from Zanthoxylum bungeanum.
Dossou Katina S S,Devkota Krishna P,Morton Cynthia,Egan Josephine M,Lu Guanghua,Beutler John A,Moaddel Ruin
Journal of natural products
In order to study cannabinoid receptor ligands, a novel plate-based assay was developed previously to measure internalization of CB1/CB2 receptors by determining the change in the intracellular levels of the radiolabeled agonists. This plate-based assay was also used for screening against complex matrices, specifically, in the present study screening for CB1/CB2 receptor activity of extracts for several species of the plant genus Zanthoxylum. The objective of this screen was to identify novel antagonists of the CB1 receptor, which simultaneously displayed agonist activity against the CB2 receptor, since compounds matching this criterion could be potential candidates for the treatment of type-1 diabetes. As a result, two Z. bungeanum extracts were deemed active, leading to the identification of eight compounds, of which compound 7 [(2E,4E,8E,10E,12E)-N-isobutyl-2,4,8,10,12-tetradecapentaenamide, γ-sanshool] was obtained as a promising lead compound.
Isolation of substances with antiproliferative and apoptosis-inducing activities against leukemia cells from the leaves of Zanthoxylum ailanthoides Sieb. & Zucc.
Chou Su-Tze,Chan Hsiu-Hui,Peng Hsin-Yi,Liou Meei-Jen,Wu Tian-Shung
Phytomedicine : international journal of phytotherapy and phytopharmacology
Extraction of the leaves of Zanthoxylum ailanthoides Sieb. & Zucc. affords extracts and four isolated compounds which exhibit activities against leukemia cells. The chloroform-soluble fraction (ZAC) of the crude extract of this plant showed cytotoxic activity against human promyelocytic leukemia (HL-60) and myelomonocytic leukemia (WEHI-3) cells with IC(50) values of 73.06 and 42.22 μg/mL, respectively. The active ZAC was further separated to yield pheophorbide-a methyl ester (1), pheophorbide-b methyl ester (2), 13(2)-hydroxyl (13(2)-S) pheophorbide-a methyl ester (3) and 13(2)-hydroxyl (13(2)-R) pheophorbide-b methyl ester (4) whose structures were confirmed by spectroscopic methods. Compounds 2-4 showed cytotoxic activities against both leukemia cells with IC(50) value in the range of 46.76-79.43 nM, whereas compound 1 exhibited only weak cytotoxic activity. The extracts and compounds 1-4 also induced apoptosis and DNA damage in leukemia cells after treatment. The results suggested that the Z. ailanthoides is biologically active against leukemia cells.
Mast cells control insulitis and increase Treg cells to confer protection against STZ-induced type 1 diabetes in mice.
Carlos Daniela,Yaochite Juliana N U,Rocha Fernanda A,Toso Vanina D,Malmegrim Kelen C R,Ramos Simone G,Jamur Maria C,Oliver Constance,Camara Niels O,Andrade Marcus V M,Cunha Fernando Q,Silva João S
European journal of immunology
Quantitative alterations in mast cell numbers in pancreatic lymph nodes (PLNs) have been reported to be associated with type 1 diabetes (T1D) progression, but their potential role during T1D remains unclear. In this study, we evaluated the role of mast cells in T1D induced by multiple low-dose streptozotocin (MLD-STZ) treatments, using two strains of mast cell-deficient mice (W/W(v) or Wsh/Wsh) and the adoptive transfer of mast cells. Mast cell deficient mice developed severe insulitis and accelerated hyperglycemia, with 100% of mice becoming diabetic compared to their littermates. In parallel, these diabetic mice had decreased numbers of T regulatory (Treg) cells in the PLNs. Additionally, mast cell deficiency caused a significant reduction in IL-10, TGF-β, and IL-6 expression in the pancreatic tissue. Interestingly, IL-6-deficient mice are more susceptible to T1D associated with reduced Treg-cell numbers in the PLNs, but mast cell transfer from wild-type mice induced protection to T1D in these mice. Finally, mast cell adoptive transfer prior to MLD-STZ administration conferred resistance to T1D, promoted increased Treg cells, and decreased IL-17-producing T cells in the PLNs. Taken together, our results indicate that mast cells are implicated in resistance to STZ-induced T1D via an immunological tolerance mechanism mediated by Treg cells.
Inhibition of nuclear import by protein kinase B (Akt) regulates the subcellular distribution and activity of the forkhead transcription factor AFX.
Brownawell A M,Kops G J,Macara I G,Burgering B M
Molecular and cellular biology
AFX belongs to a subfamily of Forkhead transcription factors that are phosphorylated by protein kinase B (PKB), also known as Akt. Phosphorylation inhibits the transcriptional activity of AFX and changes the steady-state localization of the protein from the nucleus to the cytoplasm. Our goal was threefold: to identify the cellular compartment in which PKB phosphorylates AFX, to determine whether the nuclear localization of AFX plays a role in regulating its transcriptional activity, and to elucidate the mechanism by which phosphorylation alters the localization of AFX. We show that phosphorylation of AFX by PKB occurs in the nucleus. In addition, nuclear export mediated by the export receptor, Crm1, is required for the inhibition of AFX transcriptional activity. Both phosphorylated and unphosphorylated AFX, however, bind Crm1 and can be exported from the nucleus. These results suggest that export is unregulated and that phosphorylation by PKB is not required for the nuclear export of AFX. We show that AFX enters the nucleus by an active, Ran-dependent mechanism. Amino acids 180 to 221 of AFX comprise a nonclassical nuclear localization signal (NLS). S193, contained within this atypical NLS, is a PKB-dependent phosphorylation site on AFX. Addition of a negative charge at S193 by mutating the residue to glutamate reduces nuclear accumulation. PKB-mediated phosphorylation of AFX, therefore, attenuates the import of the transcription factor, which shifts the localization of the protein from the nucleus to the cytoplasm and results in the inhibition of AFX transcriptional activity.
Insulin-like growth factor-I E-peptide activity is dependent on the IGF-I receptor.
Brisson Becky K,Barton Elisabeth R
Insulin-like growth factor-I (IGF-I) is an essential growth factor that regulates the processes necessary for cell proliferation, differentiation, and survival. The Igf1 gene encodes mature IGF-I and a carboxy-terminal extension called the E-peptide. In rodents, alternative splicing and post-translational processing produce two E-peptides (EA and EB). EB has been studied extensively and has been reported to promote cell proliferation and migration independently of IGF-I and its receptor (IGF-IR), but the mechanism by which EB causes these actions has not been identified. Further, the properties of EA have not been evaluated. Therefore, the goals of this study were to determine if EA and EB possessed similar activity and if these actions were IGF-IR independent. We utilized synthetic peptides for EA, EB, and a scrambled control to examine cellular responses. Both E-peptides increased MAPK signaling, which was blocked by pharmacologic IGF-IR inhibition. Although the E-peptides did not directly induce IGF-IR phosphorylation, the presence of either E-peptide increased IGF-IR activation by IGF-I, and this was achieved through enhanced cell surface bioavailability of the receptor. To determine if E-peptide biological actions required the IGF-IR, we took advantage of the murine C2C12 cell line as a platform to examine the key steps of skeletal muscle proliferation, migration and differentiation. EB increased myoblast proliferation and migration while EA delayed differentiation. The proliferation and migration effects were inhibited by MAPK or IGF-IR signaling blockade. Thus, in contrast to previous studies, we find that E-peptide signaling, mitogenic, and motogenic effects are dependent upon IGF-IR. We propose that the E-peptides have little independent activity, but instead affect growth via modulating IGF-I signaling, thereby increasing the complexity of IGF-I biological activity.
Protection against T1DM-Induced Bone Loss by Zinc Supplementation: Biomechanical, Histomorphometric, and Molecular Analyses in STZ-Induced Diabetic Rats.
Bortolin Raul Hernandes,da Graça Azevedo Abreu Bento João,Abbott Galvão Ururahy Marcela,Costa de Souza Karla Simone,Bezerra João Felipe,Loureiro Melina Bezerra,da Silva Flávio Santos,Marques Dáfiny Emanuele da Silva,Batista Angélica Amanda de Sousa,Oliveira Gisele,Luchessi André Ducati,Lima Valéria Morgiana Gualberto Duarte Moreira,Miranda Carlos Eduardo Saraiva,Lia Fook Marcus Vinicius,Almeida Maria das Graças,de Rezende Luciana Augusto,de Rezende Adriana Augusto
Several studies have established an association between diabetes and alterations in bone metabolism; however, the underlying mechanism is not well established. Although zinc is recognized as a potential preventive agent against diabetes-induced bone loss, there is no evidence demonstrating its effect in chronic diabetic conditions. This study evaluated the effects of zinc supplementation in a chronic (90 days) type 1 diabetes-induced bone-loss model. Male Wistar rats were distributed in three groups: control, type 1 diabetes mellitus (T1DM), and T1DM plus zinc supplementation (T1DMS). Serum biochemical analysis; tibia histomorphometric, biomechanical, and collagen-content analyses; and femur mRNA expression were evaluated. Relative to T1DM, the zinc-supplemented group showed increased histomorphometric parameters such as TbWi and BAr and decreased TbSp, increased biomechanical parameters (maximum load, stiffness, ultimate strain, and Young's modulus), and increased type I collagen content. Interestingly, similar values for these parameters were observed between the T1DMS and control groups. These results demonstrate the protective effect of zinc on the maintenance of bone strength and flexibility. In addition, downregulation of OPG, COL1A, and MMP-9 genes was observed in T1DMS, and the anabolic effects of zinc were evidenced by increased OC expression and serum ALP activity, both related to osteoblastogenesis, demonstrating a positive effect on bone formation. In contrast, T1DM showed excessive bone loss, observed through reduced histomorphometric and biomechanical parameters, characterizing diabetes-associated bone loss. The bone loss was also observed through upregulation of OPG, COL1A, and MMP-9 genes. In conclusion, zinc showed a positive effect on the maintenance of bone architecture and biomechanical parameters. Indeed, OC upregulation and control of expression of OPG, COL1A, and MMP-9 mRNAs, even in chronic hyperglycemia, support an anabolic and protective effect of zinc under chronic diabetic conditions. Furthermore, these results indicate that zinc supplementation could act as a complementary therapy in chronic T1DM.
Zanthoxylum alkylamides activate phosphorylated AMPK and ameliorate glycolipid metabolism in the streptozotocin-induced diabetic rats.
Ren Tingyuan,Zhu Yuping,Kan Jianquan
Clinical and experimental hypertension (New York, N.Y. : 1993)
This study aimed to evaluate the effects of Zanthoxylum alkylamides on the glycolipid metabolism of rats with streptozotocin (STZ)-induced diabetes. Diabetic rats were given daily oral treatments of 2, 4, or 8 mg/kg bw alkylamides for 28 days. Alkylamides significantly decreased fasting blood glucose and fructosamine content, as well as relieved organ enlargement caused by diabetes. The serum and liver triglyceride, malondialdehyde, and free fatty-acid contents of rats with STZ-induced diabetes were significantly reduced. Total cholesterol in the liver also significantly decreased. Quantitative polymerase chain reaction (Q-PCR) and Western blot detected insignificantly increased (P > 0.05) mRNA expression levels of adenosine monophosphate-activated protein kinase (AMPK) in the skeletal muscle of diabetic rats. However, AMPK and p-AMPK (Thr172) protein expression levels significantly increased. The mRNA and protein expression levels of silencing information regulator 1 significantly increased. The mRNA expression levels of acetyl-CoA-carboxylase (ACC) and protein p-ACC (Ser79) also increased. The mRNA and protein expression levels of glucose transporter type 4 (GLUT4) were significantly upregulated in the skeletal muscle cell membranes of diabetic rats. Results indicated that alkylamides activated the AMPK-signaling pathway. Thus, inhibiting ACC activity reduced fatty-acid synthesis. The rapid translocation of GLUT4 mediated increased glucose transport rate and reduced blood glucose. Therefore, alkylamides can ameliorate glucose and lipid metabolism disorders in diabetic rats by activating the AMPK pathway.
alkylamides ameliorate protein metabolism disorder in STZ-induced diabetic rats.
Ren Tingyuan,Zhu Yuping,Xia Xuejuan,Ding Yongbo,Guo Jing,Kan Jianquan
Journal of molecular endocrinology
This study aimed to evaluate the protein metabolism effect of alkylamides and to explore the potential mechanism in streptozotocin (STZ)-induced diabetic rats. Diabetic rats were orally treated with 2, 4 and 8 mg per kg bw of alkylamides daily for 28 days. Alkylamides decreased the relative weight of the liver and food intake, significantly increased the relative skeletal muscle weight and significantly decreased the blood urea nitrogen levels. Insulin, insulin-like growth factor 1, total protein (TP) and albumin (ALB), globular proteins and ALB proteins/globulin protein levels in serum significantly increased. TP, RNA content and RNA/DNA ratio significantly increased in the skeletal muscle of diabetic rats. Real-time quantitative polymerase chain reaction results indicated that alkylamides significantly increased the mRNA expression of insulin receptor (InR), IGF1 and insulin-like growth factor 1 receptor (IGF1R) in the liver and skeletal muscle. Moreover, the mRNA and protein expression levels of PI3K, PKB and mTOR significantly increased, whereas those of atrogin-1, muscle ring finger 1 and FOXO in the skeletal muscle significantly decreased. Alkylamides may advance protein synthesis by the PI3K/PKB/mTOR signalling pathway and attenuate the catabolism of protein through the ubiquitin-proteasome pathway. Therefore, it was possible that alkylamides ameliorate protein metabolism disorders in diabetic rats by activating the mTOR pathway.
Hypoglycemic effects of Zanthoxylum alkylamides by enhancing glucose metabolism and ameliorating pancreatic dysfunction in streptozotocin-induced diabetic rats.
You Yuming,Ren Ting,Zhang Shiqi,Shirima Gerald Gasper,Cheng YaJiao,Liu Xiong
Food & function
This study aimed to evaluate the hypoglycemic effect of Zanthoxylum alkylamides and explore the potential mechanism in streptozotocin (STZ)-induced diabetic rats. Diabetic rats were orally treated with 3, 6, and 9 mg per kg bw alkylamides daily for 28 days. As the alkylamide dose increased, the relative weights of the liver and kidney, fasting blood glucose, and fructosamine levels were significantly decreased. The alkylamides also significantly increased the body weight and improved the oral glucose tolerance of the rats. Likewise, the alkylamides significantly increased the levels of liver and muscle glycogen and plasma insulin. These substances further alleviated the histopathological changes in the pancreas of the diabetic rats. The beneficial effects of high-dose alkylamides showed a comparable activity to the anti-diabetic drug glibenclamide. Western blot and real-time PCR results revealed that the alkylamide treatment significantly decreased the expression levels of the key enzymes (phosphoenolpyruvate caboxykinase and glucose-6-phosphatase) involved in gluconeogenesis and increased the glycolysis enzyme (glucokinase) in the liver, and enhanced the expression levels of pancreatic duodenal homeobox-1, glucokinase, and glucose transporter 2 in the pancreas. In addition, it was also observed that the alkylamides, unlike glibenclamide, increased the transient receptor potential cation channel subfamily V member 1 and decreased cannabinoid receptor 1 expressions in the liver and pancreas. Therefore, alkylamides can prevent STZ-induced hyperglycemia by altering the expression levels of the genes related to glucose metabolism and by ameliorating pancreatic dysfunction.