依那普利对高糖诱导的大鼠肾小球系膜细胞产生TGF-β1、LN及CTGF表达的影响

目的 观察依那普利(Enalapril,Ena)对高糖诱导的大鼠肾小球系膜细胞(GMCs)转化生长因子-β1(TGF-β1)、层黏连蛋白(LN)及结缔组织生长因子(CTGF)表达的影响.方法 大鼠GMCs分别培养于正常浓度葡萄糖(5.5 mmol/L)、高浓度葡萄糖(20 mmol/L)及高糖和不同浓度(10~(-7)mol/L、10~(-6)mol/L、10~(-5)mol/L)的Ena中,同时设空白对照.分别采用放射免疫法、酶联免疫吸附实验(ELISA)检测不同时间(24、48、72 h)细胞培养上清液中TGF-β1、LN含量,采用逆转录聚合酶链反应(RT-PCR)检测72 h细胞CTGF mRNA的表达.结果 与空白对照组比较,正常糖组和高糖组TGF-β1分泌增加、LN合成增多、CTGF表达上调(P<0.001),高糖组更明显(P<0.001).与高糖组比较,高、中、低浓度的Ena干预均可逆转上述变化.结论 高糖可致GMCs分泌TGF-β1增加、合成LN增多、CTGF表达上调,Ena可减少细胞外基质(ECM)积聚,下调CTGF表达,从而有利于延缓糖尿病肾病肾小球硬化(GS)的进展.

Effect of enalapril on the production of transforming growth factor-β1, laminin and connective tissue growth factor in rat glomerular mesangial cells cultivated under high glucose

Objective To investigate the effect of enalapril on the production of transforming growth factor-β1(TGF-β1), laminin(LN) and connective tissue growth factor(CTGF) in rat glomerular mesangial cells(GMCs) cultivated under high glucose. Methods GMCs were cultured with normal concentration glucose(NG, 5.5 mmol/L), high concentration glucose(HG, 20 mmol/L), and HG plus different concentration of enalapril for different duration(24 h, 48 h, 72 h), and blank control was set up simultaneously. The contents of TGF-β1, LN in the supernatant of cultured. GMCs were detected by RIA, ELISA, respectively, and the expression of CTGF was detected by RT-PCR. Results Compared with those in blank control group, the contents of TGF-β1, LN were higher than those in NG and HG groups (P<0.001), and the expression of CTGF was up-regulated in NG and HG groups (P<0.001), HG group was more significant(P<0.001). Compared with those in HG group, HG plus high, middle, low concentration of enalapril groups had significantly lower TGF-β1, LN in the supernatant of cultured GMCs(P<0.005), and decreased the expression of CTGF. Conclusions GMCs can increase the secretion of TGF-β1, synthesis of LN, expression of CTGF induced by high glucose. Enalapril can reduce accumulation of extracelluar matrix(ECM), and down-regulate the expression of CTGF, then delay the progression of glomerular sclerosis of diabetic nephropathy.