绿原酸抑制TGF-β1诱导的人肾小管上皮细胞系HK-2纤维化因子表达

Chlorgenic acid inhibits TGF-β1-induced fibrotic factors expression of human renal tubular epithelial cell line HK-2

目的探讨绿原酸对TGF-β1诱导的人肾小管上皮HK-2细胞表达纤维化因子的变化及其分子机制。方法体外培养HK-2细胞。用CCK-8法检测细胞增殖;用免疫细胞荧光法检测collagenⅠ、collagenⅢ的表达;RT-qPCR检测纤维化相关基因mRNA的表达;用Western blot检测TLR4/NF-κB信号通路相关蛋白的表达。结果在不同时间节点(0、12、24、36、72 h)各浓度(0、20、40、80μg/mL)CGA组细胞增殖无显著差异。与对照组相比,TGF-β1处理组collagenⅠ、collagenⅢ、α-SMA、vimentin、TGF-β1、snail及slug的表达比对照组明显增加(P<0.05),但E-cadherin的表达降低(P<0.05)。CGA能够显著降低TGF-β1对上述分子表达的影响。同时,与TGF-β1处理组相比,CGA能够降低HK-2细胞TLR4、p-IκBα及p-NF-κB蛋白表达(P<0.05)。结论CGA能够抑制TGF-β1诱导的人肾小管上皮HK-2细胞表达纤维化因子的增加,其分子机制与调节TLR4/NF-κB信号通路有关。

Objective To explore the effects of Chlorgenic acid(CGA)on TGF-β1 induced the change of human renal tubular epithelial HK-2 cell fibrotic factors change and the underlying mechanism.Methods HK-2 cells were cultured in vitro.CCK-8 assay was conducted to detect cell proliferation.Immunofluorescence assay was used to detect the expression of collagenⅠand collagenⅢ.RT-qPCR was used to detect mRNA expression of HK-2 cell fibrosis related genes.Western blot assay was carried out to detect protein expression of TLR4/NF-κB pathway.Results The proliferation rate of cells in different CGA treated groups at different time points showed no differences.The expression of collagenⅠ,collagenⅢ,α-SMA,vimentin,TGF-β1,snail and slug increased while E-cadherin decreased in TGF-β1 group(P<0.05).CGA reversed these expression changes induced by TGF-β1.Further,CGA inhibited TGF-β1 induced TLR4,p-IκBαand p-NF-κB expression(P<0.05).Conclusions Chlorgenic acid can inhibit TGF-β1 induced increase of fibrotic factors expression in human renal tubular epithelial HK-2 cell through suppressing activation of TLR4/NF-κB pathway.