沉默调节蛋白1抗胰岛INS-1细胞凋亡作用的研究

石建霞 邹大进

·糖尿病基础研究·

作者单位：200433上海，第二军医大学附属长海医院内分泌科

通讯作者：邹大进，E-mail:zwid22@medmail.com.cn

【摘要】 目的 通过观察游离脂肪酸(FFA)对INS-1 细胞沉默调节蛋白1(SIRT1)、活性氧( ROS )水平和凋亡的影响，探讨FFA损害胰岛β细胞功能及SIRT1保护细胞的机制。 方法 将 INS-1细胞分为牛血清白蛋白(BSA)对照组和FFA组，作用36h 后分别检测SIRT1 mRNA水平、ROS 水平和凋亡变化。构建SIRT1过表达质粒，转染INS-1 细胞，再在上述两组条件下作用36h 后分别检测ROS 水平和凋亡变化。 结果 与BSA组比较，FFA组SIRT1 mRNA相对表达量下降(0.50±0.12 vs 1.02±0.08, P<0.01)、ROS水平明显增加(458.15±134.94 vs 132.86±51.80, P<0.01)、凋亡增加(36.55±8.16 vs 5.85±1.65 , P<0.01)。在FFA作用下，SIRT1过表达质粒转染INS-1细胞后较转染前ROS水平下降(

284.80±87.97

vs

458.15±134.94, P<0.01)，凋亡减少(18.72±7.13 vs 36.55±8.16, P<0.01)。 结论 FFA对INS-1细胞功能的损害与氧化应激有关，而SIRT1过表达可减少ROS 产生，减少凋亡，从而保护INS-1细胞功能。

doi:10.3969/j.issn.1006-6187.2011.06.017

Role of SIRT1 anti-apoptosis in pancreatic INS-1 cell

SHI Jian-xia, ZOU Da-jin.Department of Endocrinology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China

Corresponding author: ZOU Da-jin, E-mail: zwjd22@medmail.com.cn

【Abstract】 Objective To observe the effect of free fatty acid (FFA) on SIRT1 level, reactive oxygen species (ROS) and apoptosis in pancreatic INS-1 cells, also to investigate the mechanism of FFA impairing β cell and of SIRT1 protecting the cells. Methods

The INS-1 cells were cultured in FFA and control medium for 36 hours, and then the SIRT1 mRNA, ROS and apoptosis level were detected. The constructed SIRT1-overexpressing plasmid was transfected into INS-1 cell by FuGENE, and the INS-1 cell was cultured in the two group of medium, after 36 hours

, ROS and apoptosis levels were detected. Results SIRT1 mRNA level in FFA group was significantly lower than in control group(0.50±0.12 vs 1.02±0.08, P<0.01). ROS level was significantly higher in FFA group than in control group.(458.15±134.94 vs 132.86±51.80, P<0.01). The apoptosis rate was significantly

higher in FFA than in control group (36.55±8.16 vs

5.85±1.65, P<0.01). Under FFA condition, the ROS level and apoptosis rate were significantly decreased in SIRT1-overexpressing INS-1 cells as compared with control INS-1 cells(ROS:284.80±87.97 vs 458.15±134.94, P<0.01; apoptosis rate: 18.72±7.13 vs 36.55±8.16, P<0.01). Conclusion The impairment of FFA on ISN-1 cells is related to ROS. SIRT1-overexpressing of INS-1 cells can inhibit the production of ROS, decrease apoptosis, and protect INS-1 cell.

【Key words】Silent information regulator 1; Lipid toxicity; INS-1 cell; Free fatty acid; Reactive oxygen species; Apoptosis