藻黄合剂含药血清体外培养人肾小球系膜细胞结缔组织生长因子的表达

背景：结缔组织生长因子是转化生长因子β1 的下游效应因子,其异常表达在肾小球硬化及纤维化发生、发展中起重要作用。 目的：观察藻黄合剂含药血清对肾小球系膜细胞结缔组织生长因子表达的影响。 方法：血清药理学方法制备藻黄合剂低、中、高剂量,空白对照及海昆肾喜阳性对照大鼠含药血清,分别用体积分数10%各含药血清培养基孵育高糖作用下肾小球系膜细胞48 h,另设低糖组为对照。免疫细胞化学和RT-PCR方法检测各组系膜细胞结缔组织生长因子蛋白及mRNA的表达。 结果与结论：结缔组织生长因子蛋白及mRNA表达在高糖组较低糖组明显升高(P < 0.01);藻黄合剂各剂量组结缔组织生长因子蛋白及mRNA表达均低于高糖组,且具有剂量依赖性;藻黄合剂高剂量组与海昆肾喜组比较结缔组织生长因子蛋白及mRNA表达差异无显著性意义(P > 0.05)。提示高糖可诱导肾小球系膜细胞中结缔组织生长因子的表达,而藻黄合剂可抑制高糖条件下肾小球系膜细胞中结缔组织生长因子蛋白及mRNA的表达。

Effects of serum containing Zaohuang mixture on the expression of connective tissue growth factor in human mesangial cells cultured in vitro

BACKGROUND:Connective tissue growth factor (CTGF) is a kind of downstream effector of transforming growth factor-β1. The abnormal expression of CTGF plays an important role in pathological changes of diabetic kidney disease (DKD). OBJECTIVE:To investigate the effect of Zaohuang mixture contained serum (ZHM) on the expression of CTGF in human mesangial cells (HMC). METHODS:Serums containing different concentration of ZHM and haikunshenxi were prepared by serum pharmacology method. HMC were incubated in DMEM medium with 10% contained serum and high glucose for 48 hours. The level of CTGF protein and mRNA expression was measured by immunocytochemistry and RT-PCR respectively. RESULTS AND CONCLUSION:With the stimulation of high glucose, the expression of CTGF protein and mRNA was increased significantly (P < 0.01), which then were decreased by the treatment of ZHM and showed a dose-dependent manner. Compared with haikunshenxi group, CTGF protein and mRNA expression in the ZHM high-dosage group was not statistically significant (P > 0.05). The expression of CTGF in HMC may be induced by high glucose and ZHM can inhibit the expression for prevention and treatment of DKD.