FGF2/FGFR1/ERK信号通路在高糖诱导人肾小管上皮细胞转分化中的机制及肾元颗粒的干预作用

摘要

目的:探讨FGF2/FGFR1/ERK在高糖诱导人肾小管上皮细胞转分化中的信号转导机制及肾元颗粒的干预作用.方法:以高糖(30 mmol/L)诱导HK-2细胞上皮-间充质细胞转分化建立模型,分为正常组、模型组、高渗对照组、肾元颗粒组、厄贝沙坦组及ERK阻断剂组.采用CCK-8法确定大鼠血清加入浓度,Western blot及RT-PCR检测FGF2、Coll-genⅣ蛋白及mRNA表达水平,ELISA法分析各组细胞ERK及磷酸化ERK(p-ERK)水平.结果:与正常组比较,模型组FGF2、CollgenⅣ蛋白及mRNA表达显著增高(P＜0.05),p-ERK水平显著增高(P＜0.05).与模型组比较,肾元颗粒组FGF2、CollgenⅣ蛋白及mRNA表达显著降低(P＜0.05),p-ERK水平显著降低(P＜0.05).结论:肾元颗粒含药血清能通过下调高糖诱导的HK-2细胞FGF2的表达,抑制FGF2/FGFR信号转导途径从而改善肾小管上皮细胞转分化,这可能是其防治糖尿病肾病的机制之一.%Objective:To investigate the mechanism of FGF2/FGFR1/ERK signal transduction in epithelial-mesenchymal transition (EMT) of human renal tubular epithelial cells induced by high glucose and intervention effect of the serum containing Shen yuan particles.Methods:HK-2 cells EMT were induced by high glucose (30 mmol/L),and divided into normal group,model group,control group,hypertonic group,Shen yuan particles group,irbesartan group and ERK inhibitor group.In this research,CCK -8 method was used to determine the concentration of serum in rats,Western blot and RT-PCR were used in text expression levels of FGF2 and CollgenⅣ protein,while ERK and p-ERK levels in each group were analyzed by ELISA method.Results:Compared with the normal group,the expression of FGF2,p-ERK,Collgen Ⅳand its mRNA in model group were significantly increased (P ＜ 0.05),while when compared with the model group,the above indexes were significantly decreased (P ＜ 0.05).Conclusion:The serum containing Shen yuan particles can down regulate the expression of FGF2 in HK-2 cells induced by high glucose,inhibit FGF2/FGFR signal transduction pathway to improve the EMT of renal tubular epithelial cells,which may be one of the mechanisms of this drug preventing and curing diabetic nephropathy.