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目的通过观察游离脂肪酸(FFA)对INS-1细胞沉默调节蛋白1(SIRTl)、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SIRTl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土0.08,P<0.01)、ROS水平明显增加(458.15±134.94 vs132.86±51.80,P<0.01)、凋亡增加(36.55±8.16vs 5.85±1.65,P<0.01)。在FFA作用下,SIRTl过表达质粒转染INS-I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±7.13vs 36.55±8.16,P<0.01)。结论 FFA对IN S-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INS1细胞功能。
OBJECTIVE: To investigate the effect of free fatty acid (FFA) on SIRT1, reactive oxygen species (ROS) and apoptosis in INS-1 cells and to explore the mechanism of FFA damaging pancreatic β-cell function and SIRT1 protective cells. Methods INS-1 cells were divided into bovine serum albumin (BSA) control group and FFA group, and the changes of SIRT1 mRNA level, ROS level and apoptosis were detected after 36 hours. The SIRT1 overexpression plasmid was constructed and transfected into INS-1 cells. The ROS levels and apoptosis were detected after 36 hours of treatment. Results Compared with BSA group, the relative expression of SIRT1 mRNA in FFA group decreased (0.50 ± 0.12 vs 1.02 ± 0.08, P <0.01) and ROS level increased significantly (458.15 ± 134.94 vs 132.86 ± 51.80, P <0.01) (36.55 ± 8.16 vs 5.85 ± 1.65, P <0.01). Under the action of FFA, SIRT1 overexpression plasmid transfected INS-I cells decreased compared with that before transfection (284.80 ± 87.97 vs 458.15 ± 134.94, P <0.01), and apoptosis decreased (18.72 ± 7.13 vs 36.55 ± 8.16, P <0.01). Conclusions FFA may impair the function of IN S-1 cells and oxidative stress. However, overexpression of SIRT1 can reduce the production of ROS, decrease the apoptosis and protect the function of INS1 cells.