N-乙酰半胱氨酸对脂毒性条件下INS-1细胞胰岛素信号分子基因表达影响的观察  

Effect of N-acetyl-l-cysteine on the expression of insulin signal gene of INS-1 cells subjected to free fatty acid and its mechanism

作　　者：王冰 侯志强[2] 李宏亮[3] 杨文英[3] 李洪梅 

机构地区：[1]煤炭总医院内分泌科,北京100028 [2]山东省立医院急诊科 [3]卫生部中日友好医院内分泌科

出　　处：《中国糖尿病杂志》2013年第6期562-565,共4页Chinese Journal of Diabetes

摘　　要：目的观察棕榈酸(PA)对INS-1细胞胰岛素分泌功能及胰岛素信号分子基因表达的影响及抗氧化剂N-乙酰半胱氨酸(NAC)干预的效果,探讨氧化应激(OS)的作用及机制。方法 INS-1细胞分为对照(NC)组、棕榈酸(PA)组及PA+NAC(PA+NAC)组,分别用0.2 mmol/L PA和/或0.2mg/ml NAC进行干预,48h后进行相关检测:(1)行胰岛素释放试验,检测INS-1细胞胰岛素分泌功能;(2)测定细胞内硝基酪氨酸(NT)含量,评价OS水平;(3)实时荧光定量PCR反应,检测各组细胞胰岛素受体底物1(IRS-1)、胰岛素受体底物2(IRS-2)以及葡萄糖转运子2(Glut-2)基因表达的变化。结果(1)PA组IRI降低,PA+NAC组较PA组升高[(1.16±0.11)vs(0.84±0.12),P<0.05];(2)PA干预细胞内NT水平增加,NAC干预水平降低;(3)与NC相比,PA组IRS-1,IRS-2以及Glut-2mRNA表达分别降低19.9%,29.8%,42.3%。PA+NAC组以上指标表达分别较PA组增加22.4%,47.1%,80.0%。结论 FFA长期刺激可使INS-1细胞葡萄糖刺激胰岛素分泌(GSIS)功能下降,IRS-1,IRS-2以及Glut-2mRNA表达降低,NAC抗氧化干预能改善INS-1细胞胰岛素信号传导,部分逆转脂毒性导致的胰岛β细胞分泌功能异常,其机制可能与NAC纠正氧化及抗氧化失衡有关。Objective To study the effect of palrnitic acid （PA） on the insulin secretion of INS-1 cells and expression of insulin signal transduction molecules and the effect of intervention with N-acetyl-1- cysteine （NAC） for discussing the functions and mechanism of oxidative stress. Methods The INS-1 cells were divided into NC group, PA group, and PA＋NAC group, which were treated with 0. 2 mmol/L PA and/or 0. 2 mg/mL NAC respectively. After 48 hours, （1） the insulin release test was done to determine the insulin secretion in INS-1 ceils; （2） the content of nitrotyrosine in cells was measured to evaluate the oxidative stress level; and （3） the changes in gene expression of IRS-1, IRS-2, and Glut-2 were detected with real-time PCR. Results （1） the effect of PA and NAC on the insulin secretion in INS-1 cells： IS/was significantly lowered in the PA group, while it was higher in the PA＋NAC group than in the PA group [（1.16±0. il）vs （0.84±0. 12）, P〈0. 05]; （2） the effect of PA and NAC on the oxidative stress level of INS-1 ceils, the content of nitrotyrosine （NT） treated with PA was increased, while treated with NAC decreased （P〈0. 05）; （3） compared with the normal control group, the expression of IRS-1, IRS-2, and Glut-2 mRNA in the＇ PA group was all reduced, only being 19. 9%,29.8%, and 42. 3% respectively of the normal control group （all P〈0. 05）3 compared with the PAW NAC group, those were increased, being 22.4%, 47. 1%, and 80. 0%; respectively, higher than in the PA A-NAC group （all P;0. 05）. Conclusion Free fatty acid may weaken the GSIS function of INS-1 cells and lower the IRS-1, IRS-2, and Glut-2 mRNA expression. The intervention with NAC for anti-oxidation may improve the insulin transduction signals of INS-1 cells and partially correct the secretion disorders of pancreatic islet β cells caused by lipid toxicity. The mechanism may be associated with NAC correcting the imbalance between the oxidation and anti-oxidation.

关 键 词：棕榈酸 N-乙酰半胱氧酸(NAC) INS-1细胞 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]