High Passage MIN6 Cells Have Impaired Insulin Secretion with Impaired Glucose and Lipid Oxidation

Type 2 diabetes is a metabolic disorder characterized by the inability of beta-cells to secrete enough insulin to maintain glucose homeostasis. MIN6 cells secrete insulin in response to glucose and other secretagogues, but high passage (HP) MIN6 cells lose their ability to secrete insulin in response to glucose. We hypothesized that metabolism of glucose and lipids were defective in HP MIN6 cells causing impaired glucose stimulated insulin secretion (GSIS). HP MIN6 cells had no first phase and impaired second phase GSIS indicative of global functional impairment. This was coupled with a markedly reduced ATP content at basal and glucose stimulated states. Glucose uptake and oxidation were higher at basal glucose but ATP content failed to increase with glucose. HP MIN6 cells had decreased basal lipid oxidation. This was accompanied by reduced expressions of Glut1, Gck, Pfk, Srebp1c, Ucp2, Sirt3, Nampt. MIN6 cells represent an important model of beta cells which, as passage numbers increased lost first phase but retained partial second phase GSIS, similar to patients early in type 2 diabetes onset. We believe a number of gene expression changes occurred to produce this defect, with emphasis on Sirt3 and Nampt, two genes that have been implicated in maintenance of glucose homeostasis.These authors have no support or funding to report

Modulação da expressão genica e de proteinas envolvidas no mecanismo de secreção de insulina em ilhotas pancreaticas de ratos submetidos a restrição proteica

Orientadores: Everardo Magalhães Carneiro, Antonio Carlos BoscheroTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A biossíntese e secreção de insulina pelas células beta pancreáticas são reguladas por fatores humorais. estímulos nervosos e interações celulares. Alterações nutricionais durante os períodos fetais e/ou infância. induzidos por restrição protéica ou calórica. têm sido relacionadas a mudanças estruturais e nmcionais do pâncreas. Neste estudo. utilizando a técnica de "cDNA array". avaliamos a expressão gênica de ilhotas pancreáticas de ratos alimentados. durante dois meses. pós-desmame. com dieta isocalórica de baixo teor protéico (6%: LP) ou normoprotéica (17%: NP). Nossos resultados mostraram alterações na expressão de 32 genes. sendo que a maior parte das proteínas codificadas por esses genes eram relacionadas à biossíntese/secreção de insulina e remodelamento celular. Numa seglmda etapa. através de RT-PCR eAbstract: The biosynthesis and insulin secretion by pancreatic B cells are regulated by hlUllOral tàctors. nervous stimulus and intercellular interactions. Nutritional alterations during fetal and early postnatal periods. induced by either low protein or caloric restriction diets. have been related to changes in the structure and function of pancreas. ln this smdy. using cDNA array technique. we evaluated the panern 01' gene expression in pancreatic islets tTorn rats fed an isocaloric low (6%. LP) and normal (17%. NP) protein diet. after weaning. Our results show 32 genes ""ith altered expression. and the majority of them encode for proteins related to biosynthesis/insulin secretion and cellular remodeling. By RT-PCR and Westem blot. we evaluated the gene expression 01' proteins related to insulin secretion. and we also observed reduction in GLliT-2. PK.IDoutoradoFisiologiaDoutor em Biologia Funcional e Molecula

RNA sequencing of all transcripts and how islet β-cells fail

In this issue, Cnop et al. (1) report on using RNA-sequencing methodology and the response of the whole transcriptome of human islets to 48-h exposure to saturated free fatty acid (FFA) palmitate. They demonstrated that palmitate altered the expression of 1,325 genes and shifted alternate splicing of 3,525 transcripts. This follows on from a similar study by the same group on the effects on human islets of 48-h exposure to the proinflammatory cytokines interleukin-1b and interferon-g in which 3,065 (16%) of transcripts were modified and, again, alternate splicing of transcripts was commonly seen (2). However, islet b-cell failure causing diabetes, particularly type 2 diabetes, develops over years and not 48 h. So how do these technically remarkable in vitro experiments on human islets help us to understand islet b-cell failure

Islet inflammation, hemosiderosis, and fibrosis in intrauterine growth-restricted and high fat-fed sprague-dawley rats

Prenatal and postnatal factors such as intrauterine growth restriction (IUGR) and high-fat (HF) diet contribute to type 2 diabetes. Our aim was to determine whether IUGR and HF diets interact in type 2 diabetes pathogenesis, with particular attention focused on pancreatic islet morphology including assessment for inflammation. A surgical model of IUGR (bilateral uterine artery ligation) in Sprague-Dawley rats with sham controls was used. Pups were fed either HF or chow diets after weaning. Serial measures of body weight and glucose tolerance were performed. At 25 weeks of age, rat pancreases were harvested for histologic assessment. The birth weight of IUGR pups was 13% lower than that of sham pups. HF diet caused excess weight gain, dyslipidemia, hyperinsulinemia, and mild glucose intolerance, however, this was not aggravated further by IUGR. Markedly abnormal islet morphology was evident in 0 of 6 sham-chow, 5 of 8 sham-HF, 4 of 8 IUGR-chow, and 8 of 9 IUGR-HF rats (chi-square, P = 0.007). Abnormal islets were characterized by larger size, irregular shape, inflammation with CD68-positive cells, marked fibrosis, and hemosiderosis. β-Cell mass was not altered by IUGR. In conclusion, HF and IUGR independently contribute to islet injury characterized by inflammation, hemosiderosis, and fibrosis. This suggests that both HF and IUGR can induce islet injury via converging pathways. The potential pathogenic or permissive role of iron in this process of islet inflammation warrants further investigation

Fatty Acid Signaling in the Beta-Cell and Insulin Secretion

Fatty acids (FAs) and other lipid molecules are important for many cellular functions, including vesicle exocytosis. For the pancreatic β-cell, while the presence of some FAs is essential for glucose-stimulated insulin secretion, FAs have enormous capac

Circulating lipids are lowered but pancreatic islet lipid metabolism and insulin secretion are unaltered in exercise-trained female rats

Deteriorating islet β-cell function is key in the progression of an impaired glucose tolerance state to overt type 2 diabetes (T2D), a transition that can be delayed by exercise. We have previously shown that trained rats are protected from heart ischem

Decreased Insulin Secretion In Islets From Rats Fed A Low Protein Diet Is Associated With A Reduced Pkaalpha Expression.

A low protein diet has been shown to affect the amount and activity of several enzymes and to decrease insulin secretion by islets isolated from rats fed such a diet. To understand the mechanisms involved in this phenomenon, we investigated the effects of forskolin, a stimulator of adenylyl cyclase, on insulin secretion by pancreatic islets from rats fed a normal (17%; NP) or low (6%; LP) protein diet for 8 wk. Isolated islets were incubated for 1 h in Krebs-bicarbonate solution containing 8.3 mmol glucose/L, with or without 10 micromol forskolin/L. The forskolin-induced insulin secretion was higher in islets from NP rats than in those from LP rats (P<0.05). Western blotting revealed that the amount of the alpha catalytic subunit of protein kinase A (PKAalpha) was 35% lower in islets from LP rats than in islets from NP rats (P<0.05). Moreover, PKAalpha mRNA expression was reduced by 30% in islets from LP rats (P<0.05). Our results indicated a possible relationship between a low protein diet and a reduction in PKAalpha expression. These alterations in PKAalpha may be responsible in part for the decreased insulin secretion by islets from rats fed a low protein diet.13463-