能量限制诱导的Sirt1高表达对胰岛β细胞的寿命及胰岛素分泌功能的影响_医学论文.docVIP

能量限制诱导的Sirt1高表达对胰岛β细胞的寿命及胰岛素分泌功能的影响_医学论文 能量限制诱导的Sirt1高表达对胰岛β细胞的寿命及胰岛素分泌功能的影响_医学论文 作者：朱志宏，陈慎仁，傅玉才，魏炽炬，林超，杨杰华 【摘要】 目的：研究能量限制(CR)对大鼠胰岛及胰岛β瘤细胞(NIT-1细胞)寿命和胰岛素分泌功能的影响，为2型糖尿病(T2DM)的发病机制及防治提供实验依据和理论基础。方法：①18月龄健康雄性SD大鼠12只随机分为CR组和对照组，饲养6个月后取胰尾组织进行实验，用免疫组化染色检测胰岛中Sirt1和insulin表达水平，β-半乳糖苷酶(β-Gal)染色反映胰岛细胞衰老情况；②NIT-1细胞亦分为对照组和CR组；用RT-PCR、免疫细胞化学染色检测Sirt1的表达变化，用放射免疫及免疫细胞化学检测胰岛素的分泌及表达量的变化，用细胞计数法检测细胞倍增周期的变化。结果：①Sirt1在SD大鼠胰岛及NIT-1细胞的胞浆胞核中均有表达，CR组Sirt1表达量均增加；②CR可使大鼠胰岛β-Gal染色阳性率下降、NIT-1细胞倍增周期延长；③CR可使大鼠胰岛及NIT-1细胞胰岛素表达量明显减少，NIT-1细胞胰岛素分泌量亦明显减少。结论：CR通过上调Sirt1基因的表达而延缓β细胞的衰老，CR还通过减少胰岛素的表达及分泌从而减少胰岛素信号的刺激，减轻β细胞的负荷并改善胰岛素的分泌功能，因此CR有利于防止T2DM的发生发展。 【关键词】 能量限制；Sirt1；胰岛素；2型糖尿病 　　［Abstract］Objective: To investigate the effects of calorie restriction(CR)on life span and insulin secretion in the pancreatic islet of SD rat and β neoplastic cell(NIT-1 cell), and to provide some evidences for pathogenesis and prevention of type 2 Diabetes Mellitus(T2DM). Methods: ①Twelve male SD rats of 18 months old were randomized into CR and control groups. The caudal end of islet was collected after 6 months of breeding. The expressions of Sirt1 and insulin were detected by immunohistochemistry, β-galactosidase(β-Gal)was utilized to identify the senescent in pancreatic islets. ②NIT-1 cell was also divided into CR and control groups. The expression of Sirt1 was detected by RT-PCR and Immunocytochemistry, the secretion and expression of insulin were detected by radioimmunity and immunocytochemistry, the cell multiplication cycle was calculated by cytometry. Results: ①Sirt1 hyper-expression was discovered in the CR group, Sirt1 expressed both in plasma and nucleus. ②The positive ratio of β-Gal was diminised in the islet and the cell multiplication cycle was extended in NIT-1 cell by CR. ③Insulin expression was obviously decreased both in the islet and NIT-1 cells treated with CR, besides, the secretion of insulin was also decreased in NIT-1 cell. Conclusion: CR delays the aged proceeding of β cell by enhancing the expressi