普罗布考对甲基胍诱导纤维化相关因子在HK-2细胞表达的影响

Effect of probucol on the expression of fibrosis correlation factor in HK-2 cells caused by methylguanidine

目的研究降脂药普罗布考对小分子尿毒素甲基胍诱导纤维化相关因子在人肾小管上皮细胞表达的影响。方法 HK-2细胞分3组培养,A组(正常对照,不加干预因素);B组培养基中加入0.5mmol.L-1甲基胍;C组培养基中加入0.5mmol.L-1甲基胍和20μmol.L-1普罗布考。各组细胞培养48h后用免疫细胞化学、RT-PCR检测TGF-β1、CTGF和FN,并用流式细胞仪对以上因子作定量检测,以观察其在HK-2细胞中的表达。结果在甲基胍刺激后,细胞免疫化学染色,HK-2细胞质中着色,显示TGF-β1、CTGF和FN均有表达。正常对照组HK-2细胞质中无着色。测定结果显示甲基胍组TGF-β1、CTGF和FN显著高于正常对照组(P<0.01),加入普罗布考后,表达显著下降(P<0.01)。结论甲基胍能促进纤维化相关因子在肾小管上皮细胞中表达;普罗布考能抑制甲基胍诱导纤维化相关因子在肾小管上皮细胞的表达。

Objective To investigate the effect of probucol on mRNA expression of the fibrotic correlation factors on human renal tubular epithelial cell line （HK-2） with the intervention of methylguanidine.Methods HK-2 cells were treated with nothing in group A,0.5 mmol·L-1 methylguanidine in group B,and 0.5 mmol·L-1 methylguanidine and 20 μmol·L-1 probucol in group C.The protein and mRNA expression of TGF-β1,CTGF and FN were examined by immunocytochemistry,flow cytometry and RT-PCR.Results The protein and mRNA expression of TGF-β1,CTGF and FN in HK-2 cells were significantly increased when HK-2 cells were cultivated with methylguanidine（P0.01）,but when probucol was added they decreasd （P0.01）.Conclusion Methylguanidine can promote the expression of fibrosis correlation factors in renal tubular epithelial cells.Probucol can inhibit the expression of fibrosis correlation factors in renal tubular epithelial cells caused by methylguanidine.