血管紧张素Ⅱ对RIN-m细胞胰岛素基因表达的影响及其分子机制研究  

Effect of AngⅡ on Insulin Gene Expression in RIN-m Cells and Its Molecular Mechanism

作　　者：鲁辛[1] 张桦 

机构地区：[1]北京航天总医院内分泌科,北京100076 [2]广州珠江医院内分泌科

出　　处：《中国药师》2015年第12期2025-2029,共5页China Pharmacist

摘　　要：目的:探讨血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)对RIN-m细胞胰岛素基因表达的影响及其相关分子机制。方法:常规培养大鼠胰岛素瘤RIN-m细胞,分为3组:空白对照组、100 nmol·L^(-1)AngⅡ组和氯沙坦预处理组,干预24h,采用RTPCR法检测胰岛素基因表达,流式细胞仪检测2',7'-二氯荧光素(DCF)的平均荧光强度,RT-PCR法检测胰十二指肠同源盒-1(PDX-1)及肌腱膜纤维肉瘤肿瘤基因同系物A(MafA)mRNA表达,Western-blot法检测PDX-1及MafA蛋白表达。结果:100nmol·L^(-1)AngⅡ组与空白组和氯沙坦预处理组间的胰岛素mRNA表达、活性氧(reactive oxygen species,ROS)水平、PDX-1、MafA mRNA及蛋白表达均有显著差异(P<0.05),后两者之间差异无统计学意义(P>0.05)。结论:AngⅡ可能通过氧化应激途径下调β细胞的PDX-1及MafA活性,进而抑制β细胞胰岛素基因表达。氯沙坦预处理可以拮抗AngⅡ对β细胞的氧化应激损伤,从而在胰岛素基因表达方面对β细胞起到保护作用。Objective： To discuss the effect of Ang Ⅱ on insulin gene expression IN RIN-m cells and its molecular mechanism. Methods： RIN-m cells were cultured and divided into three groups, including the control group, 100 nmol L-1 Ang Ⅱ group and losartan pretreatment group. After 24-hour incubation, insulin gene expression in RIN-m cells was detected by RT-PCR, the mean flu- orescent intensity of 2-, 7-dichlorofluorescein （DCF） was detected by flow cytometry, PDX-1 and MafA mRNA expression were detected by RT-PCR and the protein expression was detected by Western-blot. Results： Insulin expression in RIN-m cells, cellular ROS level, PDX-1 expression and MafA expression in 100 nmol L-l Ang Ⅱ group were significantly different from those in the control group and losartan pretreatment group （P 〈 0.05）. The later two groups had no significant differentces in those indices （P 〉 0.05 ）. Conclusion： Ang Ⅱ can down-regulate PDX-1 and MafA expression in Is-cells through oxidative stress pathway, and then inhibit insulin gene expression. Pretreatment with losartan can antagonize the effect of Ang Ⅱ, and has protective effect on IS-cells in the aspect.of insulin gene expression.

关 键 词：血管紧张素Ⅱ RIN-M细胞 胰岛素基因表达 分子机制 

分 类 号：R966[医药卫生—药理学]