洋参御唐方对高糖诱导下人肾小球系膜细胞增殖及TIMP-1 mRNA、MMP-9 mRNA表达影响

Effect of Yangshen Yutang Formula on Proliferation and Expression of TIMP -1 mRNA and MMP -9 mRNA Induced by High Glucose in Human Glomerular Mesangial Cells

目的:通过研究洋参御唐方对高糖刺激下人肾小球系膜细胞(HMC)表达的影响,探讨其对糖尿病肾病(DN)肾脏保护的作用机制。方法:应用血清药理学方法制备洋参御唐方及贝那普利药物血清,再进行药物血清干预的研究。将体外培养的HMC分为对照组、洋参御唐方高剂量组、洋参御唐方中剂量组、洋参御糖方低剂量组、高糖组、贝那普利组共6组。于培养的24 h、48 h及72 h用MTT法检测各组细胞的增殖情况;并于相应干预因素作用48 h及72 h后,运用Real-time PCR技术法检测其相关mRNA即TIMP-1 mRNA、MMP-9 mRNA的相关表达。结果:高糖刺激能诱导TIMP-1 mRNA表达的上调、MMP-9 mRNA表达的下调;与高糖组相比,洋参御唐方高、中剂量组及贝那普利组TIMP-1 mRNA的表达下调、MMP-9 mRNA的表达上调,且以洋参御唐方高剂量组逆转TIMP-1 mRNA、MMP-9 mRNA表达失衡更为显著。结论:洋参御唐方可抑制高糖培养下HMC的增殖,且抑制效果优于贝那普利,其作用与降低TIMP-1蛋白的表达,上调MMP-9蛋白表达有关。洋参御唐方可通过逆转TIMP-1、MMP-9蛋白的表达,从而抑制高糖环境HMC的增殖可能是其降低细胞外基质堆积减轻肾间质纤维化的机制之一。

Objective:To investigate the effect of Yangshen Yutang formula on the expressions of human glomerular Mesangial cells(HMC)stimulated by high glucose,and to explore the mechanism of its effect on kidney protection of diabetic nephropathy(DN).Methods:The method of serum pharmacology was used to prepare the drug serum of Yangshen Yutang Formula and Benazepril.HMC cultured in vitro was divided into the control group,Yangshen Yutang high-dose group,Yangshen Yutang middle-dose group and Yangshen Yutang low-dose group,high glucose group and Benazepril group.MTT was used to detect cell proliferation at 24 h,48 h and 72 h in each group;the expressions of TIMP-1 mRNA and MMP-9 mRNA were detected by real-time PCR at 48 h and 72 h of the corresponding intervention in each group.Results:High glucose stimulation could induce the up-regulation of TIMP-1 mRNA and the down-regulation of MMP-9 mRNA;compared to those in the high sugar group,the expression of TIMP-1 mRNA was down-regulated and MMP-9 mRNA expression was up-regulated in the high-dose group,the middle-dose and the Benazepril group;and the expression imbalance of TIMP-1 mRNA and MMP-9 mRNA was reversed more significantly in the high-dose group.Conclusion:Yangshen Yutang Formula can inhibit HMC growth under high glucose culture,its inhibition effect is superior to that of Benazepril.The effect is related to reducing the expression of TIMP-1 protein and upregulating the expression of MMP-9 protein.Yangshen Yutang Formula can inhibit the proliferation of HMC under high glucose environment by reversing the protein expressions of TIMP-1 and MMP-9,which may be one of its mechanisms to reduce extracellular matrix accumulation to alleviate renal interstitial fibrosis.