目的:观察雷帕霉素对高糖诱导肾小球系膜细胞增生、肥大及细胞外基质合成的影响。方法:在高糖状态下培养系膜细胞,予以不同剂量雷帕霉素进行干预,采用MTT的方法观察细胞增生能力,并用流式细胞仪检测各组系膜细胞体积,以前向角散射光平均强度表示细胞大小,Western印迹法检测细胞中纤维连接蛋白(FN)及Ⅳ型胶原(ColⅣ)的表达变化。结果:同正常糖及甘露醇组比较,高糖可使肾小球系膜细胞增生且发生肥大,同时高糖环境下系膜细胞分泌的FN及ColⅣ显著增加(P〈0.05),而雷帕霉素能显著抑制高糖的上述作用(P〈0.05),且呈剂量依赖性。结论:雷帕霉素可以抑制高糖状态下系膜细胞的增生、肥大及细胞外基质的合成,且呈剂量依赖性,有望用于糖尿病肾病的防治。
Objective:To observe the changes of extracellular matrix(ECM)production and hypertrophy of glomerular mesangial cells(GMC)induced by high glucose rats,and the inhibitory role of rapamycin.Methods: GMC cultured with high glucose,and different concentration of rapamycin.Cell proliferation rate was measured by MTT.The sizes of GMC were indicated by forward scatter intensity,measured by flow cytometery,and the levels of fibronection(FN) and collagen Ⅳ(Col Ⅳ) in the GMC were detected by Western blotting.Results: Compared with normal glucose group and osmotic control group,high glucose could induce GMC hypertrophy(P〈0.05).And the levels of FN,Col Ⅳ in GMC were higher in high glucose group than those in the other two groups(P〈0.05).Rapamycin could significantly dose dependently attenuated the GMC hypertrophy(P〈0.05),and lowered ECM production of cultured GMC(P〈0.05).Conclusion:High glucose can induce hypertrophy and ECM synthesis of GMC,which are significantly inhibited by rapamycin.Therefore,rapamycin may be used in the prevention and treatment of diabetic nephropathy.