Regulation Of Pancreatic Β-cell Function Research Articles

اكتشاف جين جديد يساهم في الإصابة بمرض السكري ويؤثر في استقلاب الغلوكوز

الملخص المقدمة وهدف الدراسة: يعدّ مرض السكري من النوع الثاني مرضًا متعدد العوامل والجينات. وقد تم اكتشاف أكثر من 140 متغيرًا جينيًا وراثيًا مرتبطًا بسببية المرض، ولكنها تفسر فقط 10% من الأسباب الوراثية للسكري. لهذا، تسعى هذه الدراسة إلى استخدام نهج جديد تُحدَّد من خلاله جينات جديدة مشارِكة في فسيولوجية مرض السكري وإفراز الإنسولين. طريقة البحث: تم تحليل بيانات تسلسل الحمض النووي الريبوزي من جزر البنكرياس البشرية جُمعت من 89 متبرعًا. كما أُجريت سلسلة من التجارب الوظيفية في خلايا بيتا الحيوانية (INS-1) من الفئران للتحقق من الدور المحتمل للجين المرشح بما في ذلك تثبيط نشاطه، وإفراز الإنسولين، وحيوية الخلية، وموت الخلية المبرمج، والتعبير الجيني. النتائج: قد أظهر تحليل التعبير الجيني التفاضلي لبيانات تسلسل الحمض النووي الريبوزي أن 231 جينًا كان لها تعبير منخفض على نحو ملحوظ في جزر البنكرياس من المتبرعين المرضى بالسكري مقارنة بالأصحاء. وتم خفض عدد الجينات إلى 37 من جراء تطبيق قطع إحصائي أكثر صرامة. ومن بين تلك الجينات، تم اختيار (PPP1R1A) كجين مرشح بسبب ارتباطه القوي مع معدل السكري التراكمي، وتعبيره الجيني المرتفع في جزر البنكرياس البشرية مقارنة بباقي الجينات. وأيضًا، وُجد أن تعبير الجينات الوظيفية المهمة في خلايا بيتا مثل (INS) و(PDX1) و(GLUT2) و(INSR) انخفض على نحو كبير في الخلايا التي ثُبّط فيها نشاط (PPP1R1A). وخلصت الدراسة إلى أن (PPP1R1A) جين منظّم لوظيفة خلايا بيتا وإفراز الإنسولين، كما سلطت الضوء على قائمة من الجينات التي قد يكون لها دور وتأثير في مرض السكري. Background: Type 2 diabetes (T2D) is a multifactorial polygenic disease with an unclear causative mechanism. Close to 140 genetic variants are associated with T2D, but they only explain 10% of the heritability. Aim: In this study, we employed a novel approach to identify novel genes involved in the pathophysiology of T2D and insulin secretion. Methods: We analyzed RNA-sequencing data from human pancreatic islets at a global level. Furthermore, we performed a serial of functional experiments in rat pancreatic β-cells to validate the potential role of the candidate gene including siRNA silencing, insulin secretion, cell viability, apoptosis, QPCR, and western blot expression of the key molecule in β-cells. Results: Differential expression analysis reveals those 231 genes are significantly (P<0.05) downregulated in diabetic islets compared to nondiabetics. However, the number of genes was restricted to 37 genes by applying a cutoff of (P<0.01). Of these genes, PPP1R1A was selected as a candidate gene due to its strong correlation with HbA1c and high expression in human pancreatic islets. Silencing of PPP1R1A in INS-1 cells was markedly reduced, insulin secretion while cell viability and apoptosis were not affected. In addition, the expression of Insulin, PDX1, GLUT2, and INSR was significantly down-regulated in PPP1R1A-silenced cells. Expression of NEUROD1 and GCK was not affected. Conclusion: The study identifies PPP1R1A as a novel regulator of pancreatic β-cell function and insulin secretion. Moreover, it introduces a list of potential genes to be explored for their role in the pathogenesis of T2D.

Isoform-specific Roles of Prolyl Hydroxylases in the Regulation of Pancreatic β-Cell Function.

Pancreatic β-cells can secrete insulin via 2 pathways characterized as KATP channel -dependent and -independent. The KATP channel–independent pathway is characterized by a rise in several potential metabolic signaling molecules, including the NADPH/NADP+ ratio and α-ketoglutarate (αKG). Prolyl hydroxylases (PHDs), which belong to the αKG-dependent dioxygenase superfamily, are known to regulate the stability of hypoxia-inducible factor α. In the current study, we assess the role of PHDs in vivo using the pharmacological inhibitor dimethyloxalylglycine (DMOG) and generated β-cell-specific knockout (KO) mice for all 3 isoforms of PHD (β-PHD1 KO, β-PHD2 KO, and β-PHD3 KO mice). DMOG inhibited in vivo insulin secretion in response to glucose challenge and inhibited the first phase of insulin secretion but enhanced the second phase of insulin secretion in isolated islets. None of the β-PHD KO mice showed any significant in vivo defects associated with glucose tolerance and insulin resistance except for β-PHD2 KO mice which had significantly increased plasma insulin during a glucose challenge. Islets from both β-PHD1 KO and β-PHD3 KO had elevated β-cell apoptosis and reduced β-cell mass. Isolated islets from β-PHD1 KO and β-PHD3 KO had impaired glucose-stimulated insulin secretion and glucose-stimulated increases in the ATP/ADP and NADPH/NADP+ ratio. All 3 PHD isoforms are expressed in β-cells, with PHD3 showing the most distinct expression pattern. The lack of each PHD protein did not significantly impair in vivo glucose homeostasis. However, β-PHD1 KO and β-PHD3 KO mice had defective β-cell mass and islet insulin secretion, suggesting that these mice may be predisposed to developing diabetes.

179-OR: Defining Circadian Clock Dependency of Pancreatic Islet Transcriptional and Epigenetic Identity

The circadian regulation of pancreatic β-cell function and identity is critical for the maintenance of glucose homeostasis. The circadian clock is entrained by rhythmic environment cues (e.g., light/dark and feeding/fasting cycles) that coordinate expression of the molecular circadian machinery. Nevertheless, the relative contribution of the aforementioned components in maintaining circadian β-cell identity remains unclear. Therefore, we set out to test the hypothesis that the molecular clockwork is required for the circadian regulation of β-cell transcriptional and epigenetic identity. To address this question, we performed diurnal RNA sequencing (RNA-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) in pancreatic islets isolated from adult wild type (WT) and β-cell-specific Bmal1 (requisite circadian clock transcription factor) deficient (β-Bmal1-/-) mice. Time-dependent RNA-seq of β-Bmal1-/- islets revealed >95% decrease in the number of rhythmic transcripts compared to WT islets. Of the remaining rhythmic genes in β-Bmal1-/- islets, they predominately encoded for GO pathways regulating protein folding (FDR<.05). Correspondingly, ATAC-seq analysis found that there was >50% decrease in rhythmic chromatin accessibility in β-Bmal1-/- islets compared to WT. In line with transcriptomic findings, the remaining rhythmically accessible regions in β-Bmal1-/- islets annotated to regions regulating protein folding and insulin processing (FDR<.05). Interestingly, these regions were enriched for transcription factor motifs critical for insulin processing and biosynthesis including RFX and FOXA1. This study suggests that although the circadian clockwork is critical for entrainment of circadian β-cell identity, environmental stimuli (e.g., feeding) may maintain rhythmicity of insulin biosynthesis and protein folding in lieu of the circadian clock. Disclosure M. Brown: None. A. Matveyenko: None. Funding National Institutes of Health (DK098468 to A.M.), (DK123834 to M.B.)

Regulation of Pancreatic β-Cell Function by the NPY System.

The neuropeptide Y (NPY) system has been recognized as one of the most critical molecules in the regulation of energy homeostasis and glucose metabolism. Abnormal levels of NPY have been shown to contribute to the development of metabolic disorders including obesity, cardiovascular diseases, and diabetes. NPY centrally promotes feeding and reduces energy expenditure, while the other family members, peptide YY (PYY) and pancreatic polypeptide (PP), mediate satiety. New evidence has uncovered additional functions for these peptides that go beyond energy expenditure and appetite regulation, indicating a more extensive function in controlling other physiological functions. In this review, we will discuss the role of the NPY system in the regulation of pancreatic β-cell function and its therapeutic implications for diabetes.

Inhibitory effect of UDP-glucose on cAMP generation and insulin secretion

Type-2 diabetes (T2D) is a global disease caused by the inability of pancreatic β-cells to secrete adequate insulin. However, the molecular mechanisms underlying the failure of β-cells to respond to glucose in T2D remains unknown. Here, we investigated the relative contribution of UDP-glucose (UDP-G), a P2Y14-specific agonist, in the regulation of insulin release using human isolated pancreatic islets and INS-1 cells. P2Y14 was expressed in both human and rodent pancreatic β-cells. Dose-dependent activation of P2Y14 by UDP-G suppressed glucose-stimulated insulin secretion (GSIS) and knockdown of P2Y14 abolished the UDP-G effect. 12-h pretreatment of human islets with pertussis-toxin (PTX) improved GSIS and prevented the inhibitory effect of UDP-G on GSIS. UDP-G on GSIS suppression was associated with suppression of cAMP in INS-1 cells. UDP-G decreased the reductive capacity of nondiabetic human islets cultured at 5 mm glucose for 72 h and exacerbated the negative effect of 20 mm glucose on the cell viability during culture period. T2D donor islets displayed a lower reductive capacity when cultured at 5 mm glucose for 72 h that was further decreased in the presence of 20 mm glucose and UDP-G. Presence of a nonmetabolizable cAMP analog during culture period counteracted the effect of glucose and UDP-G. Islet cultures at 20 mm glucose increased apoptosis, which was further amplified when UDP-G was present. UDP-G modulated glucose-induced proliferation of INS-1 cells. The data provide intriguing evidence for P2Y14 and UDP-G's role in the regulation of pancreatic β-cell function.

PICK1 attenuates high glucose-induced pancreatic β-cell death through the PI3K/Akt pathway and is negatively regulated by miR-139-5p

Diabetes mellitus is a metabolic disease characterized by an increase in blood glucose levels due to lack of insulin secretion. Previous studies have confirmed that PICK1 is critical for both β-cell function and glucose homeostasis. The aim of this study was to investigate the role of PICK1 in response to high glucose-induced β-cell dysfunction and the molecular mechanism of regulation of PICK1. We found that overexpression of PICK1 in db/db diabetic mice significantly improved glucose tolerance and increased insulin release. High glucose treatment of Min6 cells inhibited PICK1 expression, and overexpression of PICK1 protected against high glucose-induced pancreatic cell dysfunction. Activation of the PI3K/Akt pathway by PICK1 in Min6 cells resulted in increased GLUT2 expression and this increase was abolished by treatment with a PI3K-specific inhibitor. Further, we showed that expression of PICK1 is negatively regulated by miR-139-5p through directly targeting its 3′UTR. These data suggested that PICK1 may participate in the functional protection of pancreatic β-cells through PI3K/Akt signaling, promote insulin secretion, and delay the progression of diabetes, and is negatively regulated by miR-139-5p, further clarifying the regulation of pancreatic β-cell function.

THE IMPACT OF DIFFERENT REGIMENS OF VITAMIN D3 ON GLUCOSE HOMEOSTASIS IN TYPE 2 DIABETIC PATIENTS

Objective: Vitamin D has a role in the regulation of pancreatic β-cell function and insulin sensitivity. Accordingly, Vitamin D deficiency is considered to be a risk factor for the development of type 2 diabetes mellitus (T2DM) and its complications. Therefore, the aim of the study was to assess and compare the effect of different regimens of Vitamin D3 on glucose homeostasis in patients with T2DM.
 Methods: The study included 80 patients with T2DM taking oral antidiabetic drugs. The patients were randomized to receive antidiabetic drugs alone or with different regimens of Vitamin D3 for 3 months. Vitamin D3-treated patients were supplemented by either daily oral 4000 IU Vitamin D3, weekly oral 50,000 IU Vitamin D3, or a single parenteral dose of 300,000 IU Vitamin D3. In addition to the assessment of patient characteristics, laboratory measurements of serum creatinine, blood urea, total and ionized calcium, serum phosphorus, fasting blood glucose, fasting serum insulin, homeostasis model assessment of insulin resistance, hemoglobin A1c, and 25(OH) Vitamin D levels were measured at the beginning and after 3 months.
 Results: After 3 months, the increased Vitamin D levels resulting from the daily and weekly oral doses of Vitamin D3 caused a significant decrease in metabolic parameters, whereas the parenteral dose demonstrated a non-significant decrease.
 Conclusion: Oral daily and weekly doses of Vitamin D3 could improve glucose homeostasis equally in patients with T2DM and better than a single parenteral dose of Vitamin D3.

Oleic acid increases the transcriptional activity of FoxO1 by promoting its nuclear translocation and β-catenin binding in pancreatic β-cells

In the setting of metabolic overload, chronic elevations of free fatty acids in blood and tissues are associated with pancreatic β-cell lipotoxicity and failure. Ultimately, obesity combined with insulin resistance increases the dysfunctional demand of β-cells and contributes to the development of type 2 diabetes. Forkhead box O1 (FoxO1) is a potent transcriptional regulator of pancreatic β-cell function and tolerance to lipid stress. The present study examined the effects of stearoyl-CoA desaturase 1 (SCD1)-metabolized precursors and products, notably oleic acid, on the compensatory capacity of β-cells and their relationship with regulation of the FoxO1 and Wnt pathways. The trioleate-induced compromise of insulin sensitivity blunted the compensatory response of pancreatic β-cells in primary rat islets. These events were associated with increases in the nuclear accumulation and transcriptional activity of FoxO1. Such effects were also observed in INS-1E cells that were subjected to oleate treatment. The overexpression of human SCD1 that was accompanied by endogenously generated oleic acid also led to an increase in the nuclear abundance of FoxO1. The mechanism of the oleate-mediated subcellular localization of FoxO1 was independent of the fatty acid receptor GPR40. Instead, the mechanism involved diversion of the active β-catenin pool from an interaction with transcription factor 7-like 2 toward FoxO1-mediated transcription in β-cells. Our findings identify a unique role for oleic acid in the compensatory response of pancreatic β-cells and emphasize the importance of FoxO1 in β-cell failure in obesity-induced insulin resistance.

2177-P: Role of De Novo Sphingolipid Metabolites in Oleate-Induced Pancreatic ß-Cell Proliferation in Rats

Fatty acids (FA) are major regulators of pancreatic β-cell function. In a rat model of nutrient excess we previously showed that FA potentiate glucose-induced β-cell proliferation. Sphingolipids, derived from the intracellular metabolism of FA, act as cellular mediators in the regulation of pancreatic β-cell function. However, the contribution of sphingolipid species to FA-induced β-cell proliferation is unknown. Objective: To determine the role of de novo sphingolipid synthesis in FA-induced β-cell proliferation. Methods: Isolated rat islets were exposed to oleate or palmitate (0.5 mM) in the presence of 16.7 mM glucose for 48h. Sphingosine kinase (SphK) 1 and 2 expression was measured by real-time PCR. Serine palmitoyl transferase (SPT), SphK, and the S1P3 receptor were inhibited with Myriocin, SKI II and TY52156, respectively. β-cell proliferation was assessed in cryosections by immunohistochemical detection of insulin and the proliferation marker Ki67 or by flow cytometry by labeling for C-peptide and EdU incorporation. Wistar rats were infused for 72 hours with glucose and a lipid emulsion (CLI) to induce β-cell proliferation. Results: The monounsaturated FA oleate, but not the saturated FA palmitate, increased β-cell proliferation. Blocking de novo sphingolipid synthesis by SPT inhibition decreased oleate-induced β-cell proliferation as did inhibition of SphK and S1P3 receptor activation. Sphk1/2 mRNA levels in islets were not significantly changed following nutrient infusion in rats or FA exposure ex vivo. Conclusion: β-cell proliferation in response to oleate requires de novo sphingolipid synthesis and S1P3 signaling. Analyses are underway to assess the contribution of SphK products sphingosine-1-phosphate and dihydrosphinganine-1-phosphate in S1P3-mediated β-cell proliferation. Disclosure A. Castell: None. A. Vivoli: None. V.S. Moullé: None. J. Ghislain: None. V. Poitout: None. Funding National Institutes of Health; Canadian Institutes of Health Research

Methods to Study Roles of β-Arrestins in the Regulation of Pancreatic β-Cell Function.

Novel findings reveal important functional roles for β-arrestin 1 and β-arrestin 2 in the regulation of insulin secretion, β-cell survival, and β-cell mass plasticity not only by glucose but also by G-protein-coupled receptors, such as the glucagon-like peptide-1 (GLP-1) and the pituitary adenylate cyclase-activating polypeptide (PACAP) receptors or GPR40, or tyrosine kinase receptors, such as the insulin receptor. Here, we describe experimental protocols to knock down β-arrestins by small interference RNA, to follow subcellular localization of β-arrestins in the cytosol and nucleus of the insulinoma INS-1E rat pancreatic β-cell line, and to analyze β-arrestin protein expression by Western blot using INS-1E cells and isolated mouse or human pancreatic islets. We also provide details on how to genotype β-arrestin 2 knockout (Arrb2-/-) mice and to evaluate β-arrestin-mediated roles in β-cell mass plasticity and β-cell signaling using immunocytochemistry on pancreatic sections or on primary dispersed β-cells from wild-type mice and Arrb2-/- mice.

Arginase-II Promotes Tumor Necrosis Factor-α Release From Pancreatic Acinar Cells Causing β-Cell Apoptosis in Aging

Aging is associated with glucose intolerance. Arginase-II (Arg-II), the type-II L-arginine-ureahydrolase, is highly expressed in pancreas. However, its role in regulation of pancreatic β-cell function is not known. Here we show that female (not male) mice deficient in Arg-II (Arg-II-/-) are protected from age-associated glucose intolerance and reveal greater glucose induced-insulin release, larger islet size and β-cell mass, and more proliferative and less apoptotic β-cells compared with the age-matched wild-type (WT) controls. Moreover, Arg-II is mainly expressed in acinar cells and is upregulated with aging, which enhances p38 mitogen-activated protein kinase (p38 MAPK) activation and release of tumor necrosis factor-α (TNF-α). Accordingly, conditioned medium of isolated acinar cells from old WT (not Arg-II-/-) mice contains higher TNF-α levels than the young mice and stimulates β-cell apoptosis and dysfunction, which are prevented by a neutralizing anti-TNF-α antibody. In acinar cells, our study demonstrates an age-associated Arg-II upregulation, which promotes TNF-α release through p38 MAPK leading to β-cell apoptosis, insufficient insulin secretion, and glucose intolerance in female rather than male mice.

Regulation of pancreatic β-cell function and mass dynamics by prostaglandin signaling.

Prostaglandins (PGs) are signaling lipids derived from arachidonic acid (AA), which is metabolized by cyclooxygenase (COX)-1 or 2 and class-specific synthases to generate PGD2, PGE2, PGF2α, PGI2 (prostacyclin), and thromboxane A2. PGs signal through G-protein coupled receptors (GPCRs) and are important modulators of an array of physiological functions, including systemic inflammation and insulin secretion from pancreatic islets. The role of PGs in β-cell function has been an active area of interest, beginning in the 1970s. Early studies demonstrated that PGE2 inhibits glucose-stimulated insulin secretion (GSIS), although more recent studies have questioned this inhibitory action of PGE2. The PGE2 receptor EP3 and one of the G-proteins that couples to EP3, GαZ, have been identified as negative regulators of β-cell proliferation and survival. Conversely, PGI2 and its receptor, IP, play a positive role in the β-cell by enhancing GSIS and preserving β-cell mass in response to the β-cell toxin streptozotocin (STZ). In comparison to PGE2 and PGI2, little is known about the function of the remaining PGs within islets. In this review, we discuss the roles of PGs, particularly PGE2 and PGI2, PG receptors, and downstream signaling events that alter β-cell function and regulation of β-cell mass.

Study on Glycemic Control by Functional Foods through Regulation of Pancreatic β-Cell Function

Study on Glycemic Control by Functional Foods through Regulation of Pancreatic β-Cell Function

MicroRNAs in Pancreatic β-Cell Physiology.

The β-cells within the pancreas are responsible for production and secretion of insulin. Insulin is released from pancreatic β-cells in response to increasing blood glucose levels and acts on insulin-sensitive tissues such as skeletal muscle and liver in order to maintain normal glucose homeostasis. Therefore, defects in pancreatic β-cell function lead to hyperglycemia and diabetes mellitus. A new class of molecules called microRNAs has been recently demonstrated to play a crucial role in regulation of pancreatic β-cell function under normal and pathophysiological conditions. miRNAs have been shown to regulate endocrine pancreas development, insulin biosynthesis, insulin exocytosis, and β-cell expansion. Many of the β-cell enriched miRNAs have multiple functions and regulate pancreas development as well as insulin biosynthesis and exocytosis. Furthermore, several of the β-cell specific miRNAs have been shown to accumulate in the circulation before the onset of diabetes and may serve as potential biomarkers for prediabetes. This chapter will focus on miRNAs that are enriched in pancreatic β-cells and play a critical role in modulation of β-cell physiology and may have clinical significance in the treatment of diabetes.

Minireview: microRNA function in pancreatic β cells.

MicroRNAs are small noncoding ribonucleotides that regulate mRNA translation or degradation and have major roles in cellular function. MicroRNA (miRNA) levels are deregulated or altered in many diseases. There is overwhelming evidence that miRNAs also play an important role in the regulation of glucose homeostasis and thereby may contribute to the establishment of diabetes. MiRNAs have been shown to affect insulin levels by regulating insulin production, insulin exocytosis, and endocrine pancreas development. Although a large number of miRNAs have been identified from pancreatic β-cells using various screens, functional studies that link most of the identified miRNAs to regulation of pancreatic β-cell function are lacking. This review focuses on miRNAs with important roles in regulation of insulin production, insulin secretion, and β-cell development, and will discuss only miRNAs with established roles in β-cell function.

Long-term disruption of maternal glucose homeostasis induced by prenatal glucocorticoid treatment correlates with miR-29 upregulation

Excess of glucocorticoids (GCs) during pregnancy is strongly associated with the programming of glucose intolerance in the offspring. However, the impact of high GC levels on maternal metabolism is not clearly documented. This study aimed to test the hypothesis that mothers exposed to elevated levels of GCs might also display long-term disturbances in glucose homeostasis. Dexamethasone (DEX) was administered noninvasively to the mothers via drinking water between the 14th and the 19th days of pregnancy. Mothers were subjected to glucose and insulin tolerance tests at 1, 2, 3, 6, and 12 mo postweaning. Pregnant rats not treated with DEX and age-matched virgin rats were used as controls. Pancreatic islets were isolated at the 20th day of pregnancy and 12 mo postweaning in order to evaluate glucose-stimulated insulin secretion. The expression of the miR-29 family was also studied due to its responsiveness to GCs and its well-documented role in the regulation of pancreatic β-cell function. Rats treated with DEX during pregnancy presented long-term glucose intolerance and impaired insulin secretion. These changes correlated with 1) increased expression of miR-29 and its regulator p53, 2) reduced expression of syntaxin-1a, a direct target of miR-29, and 3) altered expression of genes related to cellular senescence. Our data demonstrate that the use of DEX during pregnancy results in deleterious outcomes to the maternal metabolism, hallmarked by reduced insulin secretion and glucose intolerance. This maternal metabolic programming might be a consequence of time-sustained upregulation of miR-29s in maternal pancreatic islets.

Diabetes mellitus is associated with an increased expression of resistin in human pancreatic islet cells

The pattern of distribution of resistin in the pancreas of diabetic patients was investigated to determine whether diabetes mellitus influences the expression of resistin. Pancreatic tissue samples retrieved, during pancreatectomy for pancreatic cancer, from cancer patients with and without type 2 diabetes were processed for immunohistochemistry. The pancreatic tissue samples were retrieved from non-cancerous and clear margins. An immunofluorescence technique was used to examine the expression of resistin and its co-localization with insulin and glucagon in pancreatic islet cells. Resistin was observed in many cells located in the central region of pancreatic islet. The expression of resistin increased significantly (p < 0.0001) in pancreatic islet cells of type 2 diabetic patients compared to control. Resistin co-localized with insulin but not glucagon in pancreatic islet cells of both normal and diabetic patients. However, the degree of co-localization was higher in pancreata of diabetic patients compared to normal. The number of human pancreatic islet cells expressing resistin increased significantly after the onset of type 2 diabetes. In conclusion, resistin may play a role in the regulation of pancreatic β-cell function.

Roux-en-Y Gastric Bypass-Induced Improvement of Glucose Tolerance and Insulin Resistance in Type 2 Diabetic Rats Are Mediated by Glucagon-Like Peptide-1

The aim of this study was to investigate the effects of Roux-en-Y gastric bypass (RYGB) on glucose tolerance and insulin resistance in type 2 diabetic rats and the possible mechanisms involved in this process. Thirty Goto-Kakizaki (GK) rats were randomly divided into three groups: RYGB operation, sham operation, and food restriction groups. Ten Wistar rats were used as non-diabetic control. The body weight and food consumption of rats were recorded 1 week before or every week after surgery. The fasting blood sugar and oral glucose tolerance test were performed using blood glucose meter. The levels of plasma insulin or glucagon-like peptide-1 (GLP-1) were evaluated by enzyme-linked immunosorbent assay. The insulin resistance was quantified using homeostasis model assessment method. The expression of GLP-1 receptor, Bcl-2, Bax, and caspase-3 was determined by Western blotting. Our results revealed that RYGB efficiently improved both glucose tolerance and insulin resistance in GK diabetic rats by upregulating GLP-1/GLP-1R expression. In addition, GLP-1R agonist exendin-4 dose-dependently increased insulin secretion in RIN-m5F cells and regulated the proliferation and apoptosis of these cells. RYGB provides a valuable therapeutic option for patients with type 2 diabetes. GLP-1 may contribute to the regulation of pancreatic β-cell function through its receptor following RYGB.

Soy genistein prevents streptozotocin‐induced diabetes through regulation of pancreatic beta‐cell function

Loss of β‐cell mass and function are central to the development of diabetes. In the present study, we found that soy genistein induced clonal and human pancreatic β‐cell proliferation, with 5 µM genistein inducing a maximal 41% increase. The effect of genistein on cell proliferation was neither dependent on estrogen receptors, nor shared by a host of structurally related compounds. Pharmacological or molecular intervention of PKA or ERK1/2 completely abolished the genistein‐stimulated proliferation of INS1 cells and human islets, suggesting that both molecules are essential for genistein action. Consistently, genistein activated cAMP/PKA and ERK1/2 signaling in β‐cells. Furthermore, we found that genistein induced protein expression of cyclin D1, a major cell‐cycle regulator essential for β‐cell growth. We also show that genistein promoted viability, reduced caspase‐3 activity and induced anti‐apoptotic protein Bcl‐2 expression in islet cells. Dietary genistein intake increased plasma levels of insulin and ameliorated hyperglycemia in Streptozotocin (STZ) induced diabetic mice. These effects were associated with improved islet mass and protected islets from STZ‐induced apoptosis in these mice. These findings demonstrate that genistein may be a novel regulator of pancreatic β‐cell function via activation of multiple intracellular signaling pathways, thereby providing an anti‐diabetic effect.

Role of the Rho-ROCK (Rho-Associated Kinase) Signaling Pathway in the Regulation of Pancreatic β-Cell Function

Extracellular matrix has a beneficial impact on beta-cell spreading and function, but the underlying signaling pathways have yet to be fully elucidated. In other cell types, Rho, a well-characterized member of the family of Rho GTPases, and its effector Rho-associated kinase (ROCK), play an important role as downstream mediators of outside in signaling from extracellular matrix. Therefore, a possible role of the Rho-ROCK pathway in beta-cell spreading, actin cytoskeleton dynamics, and function was investigated. Rho was inhibited using a new cell-permeable version of C3 transferase, whereas the activity of ROCK was repressed using the specific ROCK inhibitors H-1152 and Y-27632. Inhibition of Rho and of ROCK increased spreading and improved both short-term and prolonged glucose-stimulated insulin secretion but had no impact on basal secretion. Inhibition of this pathway led to a depolymerization of the actin cytoskeleton. Furthermore, the impact of the inhibition of ROCK on stimulated insulin secretion was acute and reversible, suggesting that rapid signaling such as phosphorylation is involved. Finally, quantification of the activity of RhoA indicated that the extracellular matrix represses RhoA activity. Overall these results show for the first time that the Rho-ROCK signaling pathway contributes to the stabilization of the actin cytoskeleton and inhibits glucose-stimulated insulin secretion in primary pancreatic beta-cells. Furthermore, they indicate that inhibition of this pathway might be one of the mechanisms by which the extracellular matrix exerts its beneficial effects on pancreatic beta-cell function.