降糖益肾方对高胰岛素诱导的人肾小球系膜细胞增殖及IRS-1、PI-3K蛋白表达的影响  

Effect of Jiangang Yishen Recipe on High Insulin Induced Cell Proliferation of Human Glomerular Mesangial Cells and the Expression of Insulin Receptor Substrate 1 and Phosphatidylinositol-3-kinase

作　　者：吴勇军[1] 喻嵘[2] 成细华[3] 吴慧[2] 伍参荣[3] 魏冠德[2] 张琴[2] 

机构地区：[1]湖南中医药大学药学院,长沙410208 [2]湖南中医药大学中医内科学重点实验室,长沙410208 [3]湖南中医药大学细胞生物学与分子技术重点实验室,长沙410208

出　　处：《中国中西医结合杂志》2014年第5期597-601,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81011120382;No.81102593;No.81273753);湖南省自然科学基金资助项目(No.12JJ2048);湖南省自然科学省市联合基金项目(No.12JJ9031);湖南省大学生研究性学习和创新性实验计划项目(No.2010-170);湖南省普通高等学校科学研究项目(No.11C0964)

摘　　要：目的观察降糖益肾方对高胰岛素诱导的人肾小球系膜细胞(human glomerular mesangial cells,HMCs)增殖及其胰岛素受体底物-1(insulin receptor substrate 1,IRS-1)、磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI-3K)蛋白表达的影响。方法将HMCs分为正常空白对照组(空白组)、高胰岛素模型组、抑制剂组、中药组。首先预实验确定胰岛素、抑制剂和降糖益肾方用药浓度。各组分别加入不同培养液,空白组予RPMI1640培养液,其余3组均予含100nmol/L胰岛素的培养液,培养24h,同时抑制剂组、中药组分别予80μmol/L LY294002、125mg/L降糖益肾方干预,继续培养48h。采用MTT法及流式细胞技术检测HMC增殖,采用免疫组化和Western blot技术检测IRS-1、PI-3K蛋白表达。结果在高胰岛素诱导下,中药组和抑制剂组HMC增殖均显著降低,IRS-1、PI-3K蛋白表达水平下调(P<0.01);与抑制剂组比较,中药组IRS-1、PI-3K蛋白表达水平下调稍弱(P<0.05)。结论降糖益肾方具有降低高胰岛素诱导的HMC增殖的作用,其机制可能与干预胰岛素信号通路有关。Objective To investigate the effect of Jiangtang Yishen Recipe （JTYSR） on high insulin induced cell proliferation of human glomerular mesangial cells （HMCs） and the expression of insulin receptor substrate 1 （ IRS-1 ） and phosphatidylinositol-3-kinase （PI-3K）. Methods HMCs were divided into 4 groups, i.e., the negative control group, the high insulin modeJ group, the JTYSR group, and the LY294002 group. The concentration of insulin, JTYSR, and LY294002 was respectively confirmed by preexperiment. Different culture solution was respectively added for different groups. RPMI1640 culture solution was added for HMCs in the negative control group, while HMCs in the rest 3 groups were cultured by 100 nmol/L insulin for 24 h. Meanwhile, HMCs from the JTYSR group and the LY294002 group were exposed to 125 mg/L JTYSR and 80 μmol/L LY294002 respectively for further 48 h. The proliferation of HMCs was detected by MTT and flow cytometry. The protein expression of IRS-1 and PI-3K in HMC was detected by immunohistochemical assay and Western blot. Results The proliferation of HMCs induced by high insulin could be significantly lowered, and the protein expression of IRS-1 and PI-3K could be down-regulated in the JTYSR group and the LY294002 group （P 〈0.01 ）. Compared with the LY294002 group, the protein expression of IRS-1 and PI-3K could be slightly down-regulated in the JTYSR group （P 〈0.05）. Conclusion JTYSR could lower high insulin induced proliferation of HMCs, and its mechanism might be related to insulin signaling pathway.

关 键 词：降糖益肾方 肾小球系膜细胞 胰岛素受体底物-1 磷脂酰肌醇-3-激酶 

分 类 号：R285.5[医药卫生—中药学]