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目的:观察糖肾颗粒对高糖诱导的大鼠肾小球系膜细胞(GMCs)表达中期因子(MK)的影响。方法:制备不同剂量糖肾颗粒含药血清和贝那普利,加入到高糖诱导的体外培养的GMCs中。采用ELISA(24、48、72h)检测细胞培养上清液中MK含量,分别采用RT-PCR法、Western Blot法检测48h细胞MK mRNA及其蛋白表达。结果:与正常对照组比较,高糖组MK分泌增加、MK mRNA及其蛋白表达上调(P<0.001)。与高糖组比较,贝那普利和糖肾颗粒两种含药血清均能逆转上述变化,后者的抑制作用强于前者(P<0.05),且抑制作用呈一定的时间、剂量依赖关系。结论:高糖可致GMCs分泌MK增加、MK mRNA及其蛋白表达上调,糖肾颗粒可减少MK分泌,下调MK mRNA及其蛋白表达,从而有利于延缓糖尿病肾病肾小球硬化的进展。
Objective: To observe the effects of Tangshen Granule on the expression of midkine (MK) induced by high glucose in rat mesangial cells (GMCs). Methods: Different doses of Tangshen Granule containing serum and benazepril were prepared and added into GMCs induced by high glucose. The content of MK in cell culture supernatant was detected by ELISA (24,48,72h). The mRNA and protein expression of MK in 48h were detected by RT-PCR and Western Blot respectively. Results: Compared with the normal control group, the MK secretion increased and the expression of MK mRNA and its protein increased (P <0.001). Compared with the high glucose group, both serums of benazepril and Tangshen Granules could reverse the above changes, the inhibition of the latter was stronger than the former (P <0.05), and the inhibitory effect was in a time-and dose-dependent manner. CONCLUSION: High glucose can increase the secretion of MK and increase the expression of MK mRNA and its protein in GMCs. Tangshen Granule can reduce the secretion of MK and down-regulate the expression of MK mRNA and its protein, which can delay the progression of glomerulosclerosis in diabetic nephropathy.