柚皮苷抑制纤维化相关蛋白表达改善大鼠肾细胞纤维化

Naringin Inhibiting Fibrosis in Rat Renal Cells by Inhibiting Fibrosis-Related Proteins

目的:探讨柚皮苷(naringin,NAR)对转化生长因子-β(transforming growth factor-β,TGF-β)诱导的肾纤维化的作用。方法:采用不同浓度TGF-β1诱导正常大鼠肾细胞构建肾纤维化细胞模型,根据α-平滑肌肌动蛋白(α-SMA)的转录水平确定TGF-β1的最佳诱导浓度;将细胞分为正常组、TGF-β1组、TGF-β1+NAR(1μmol·L^(-1))组、TGF-β1+NAR(10μmol·L^(-1))组、TGF-β1+NAR(40μmol·L^(-1))组,细胞处理完成后,采用细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞活力;显微镜观察法观察细胞形态变化;免疫荧光法检测结缔组织生长因子(connective tissue growth factor,CTGF)和纤维连接蛋白(fibronectin1,FN1)表达;Western Blot法检测Ⅰ型胶原蛋白(CollagenⅠ)的表达。结果:5μg·L^(-1) TGF-β1为肾纤维化细胞模型建立的最佳诱导浓度。CCK-8检测显示,NAR可抑制肾纤维化模型细胞异常增殖。与正常组比较,TGF-β1组CTGF、FN1、CollagenⅠ表达水平显著升高(P<0.05),细胞形态发生变化,出现纤维化;与TGF-β1组比较,NAR给药组CTGF、FN1蛋白表达水平显著降低(P<0.05),CollagenⅠ表达水平呈剂量依赖性降低(P<0.05),细胞纤维化得到改善。结论:柚皮苷可通过抑制CTGF、FN1和CollagenⅠ蛋白表达改善TGF-β1诱导的肾纤维化。

Objective:To investigate the effect of naringin(NAR)on renal fibrosis induced by transforming growth factor-β(TGF-β).Methods:The renal fibrosis cell model was established by using normal rat kidney cells induced by TGF-β1 at different concentrations,and the optimal inducing concentration of TGF-β1 was determined according to the transcription level ofα-smooth muscle actin(α-SMA).The cells were divided into the normal group,TGF-β1 group,TGF-β1+NAR(1μmol·L^(-1))group,TGF-β1+NAR(10μmol·L^(-1))group,TGF-β1+NAR(40μmol·L^(-1))group.After the cell treatment was completed,the cell viability was detected by cell counting kit-8(CCK-8).The changes in cell morphology were observed by microscopy.And the expressions of connective tissue growth factor(CTGF)and fibronectin1(FN1)were detected by immunofluorescence.The expression of CollagenⅠwas detected by the Western-blot method.Results:5μg·L^(-1) TGF-β1 was the best inducing concentration for the establishment of the renal fibrosis cell model.CCK-8 assay showed that NAR could inhibit the abnormal proliferation of renal fibrosis model cells.Compared with the normal group,the expression levels of CTGF,FN1,and CollagenⅠin the TGF-β1 group were significantly increased(P<0.05),and the cell morphology changed and fibrosis appeared.Compared with the TGF-β1 group,the protein expression levels of CTGF and FN1 in the NAR administration group were significantly decreased(P<0.05),and the expression level of CollagenⅠdecreased in a dose-dependent manner(P<0.05),and the cell fibrosis was improved.Conclusion:Naringin can improve TGF-β1-induced renal fibrosis by inhibiting the expression of CTGF,FN1,and CollagenⅠproteins.