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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 observe the effects of Jiang Tang Yishen Recipe on hyperinsulin-induced proliferation of human glomerular mesangial cells (HMCs) and the effects of insulin receptor substrate 1 (IRS-1), phosphatidyl Effect of phosphatidylinositol-3-kinase (PI-3K) protein expression. Methods HMCs were divided into normal control group (blank group), high insulin model group, inhibitor group and traditional Chinese medicine group. First pre-experiment to determine the insulin, inhibitors and hypoglycemic agents use concentration. Each group was added with different culture medium, the blank group to RPMI1640 culture medium, the remaining three groups were given 100nmol / L insulin medium, cultured for 24h, while the inhibitor group, the Chinese medicine group were 80μmol / L LY294002, 125mg / L Intervention of sugar kidney, continue to train 48h. The proliferation of HMCs was detected by MTT assay and flow cytometry. The expressions of IRS-1 and PI-3K proteins were detected by immunohistochemistry and Western blot. Results Compared with the inhibitor group, the expression of IRS-1 and PI-3K protein in HMC of Chinese medicine group and inhibitor group were significantly decreased (P <0.01) 3K protein expression decreased slightly (P <0.05). Conclusion Jiangtangyishen Decoction can reduce the hyperinsulin-induced proliferation of HMC, the mechanism may be related to the intervention of insulin signaling pathway.