水飞蓟宾对转化生长因子β1诱导的人肾小管上皮细胞中结缔组织生长因子及Ⅲ型胶原表达的影响

The effect of silibinin on the expression of connective tissue growth factor(CTGF)and the type Ⅲ collagen in HK-2 cells induced by TGF-β_1

目的:了解水飞蓟宾(silibinin)对转化生长因子(TGF)-β_1诱导的人近曲肾小管上皮细胞(HK-2)结缔组织生长因子(CTGF)表达及Ⅲ型胶原分泌的影响。方法:将体外培养的HK-2细胞分为3组:①对照组;②TGF-β_1组(TGF-β_1的终浓度为5 ng/ml);③TGF-pβ_1+水飞蓟宾组(TGF-β_1的终浓度为5 ng/ml,水飞蓟宾终浓度分别为5、10、20μg/m1)。48 h后,采用逆转录聚合酶链反应(RT-PCR)检测细胞内CTGFmRNA的水平,ELISA方法检测培养上清中胶原Ⅲ及CTGF的浓度。结果:与对照组比较,TGF-β_1刺激使HK-2细胞CTGF基因表达和蛋白分泌及Ⅲ型胶原分泌均显著增加(P<0.01)。与TGF-β_1组比较,加用5～20μg/ml水飞蓟宾可以抑制TGF-β_1刺激所诱导的CTGF基因表达和蛋白分泌及Ⅲ型胶原分泌(P<0.01),以20μg/ml水飞蓟宾效果最显著。结论:水飞蓟宾能部分抑制TGF-β_1诱导CTGF的基因和蛋白的表达及Ⅲ型胶原分泌,提示水飞蓟宾可能通过阻抑CTGF表达及减少细胞外基质的积聚,具有潜在的抗肾纤维化作用。

Objective：To investigate the effect of silibinin on the expression of connective tissue growth factor （CTGF） and the secretion of type Ⅲ collagen in HK-2 cells induced by TGF-β1. Methods：In vitro,HK-2 cells were divided into three groups： （1）ontrol group; （2）5 ng/ml TGF-β1, group; （3）5 ng/ml TGF-β1＋silibinin with different concentrations group, 5μg/ml, 10μg/ml and 20μg/ml, respectively. 48 h later,the expression of CTGF mRNA in cells were detected by reverse transcription-polymerase chain reaction（RT- PCR） ;the concentrations of CTGF and collagen Ⅲ secreted into the culture supernatant was determined by ELISA. Results：Compared with control group,the expression of CTGF mRNA and the concentration of the secreted collagen Ⅲ and CTGF were all increased markedly in the group of TGF-β1 treatment （P 〈 0.01）. Compared with TGF-β1 group,the different concentrations of （5μg/ml, 10μg/ml and 20 μg/ml） silibinin could also effectively suppress the expressions of CTGF mRNA and the concentration of the secreted col lagen Ⅲ and CTGF （P 〈 0.01 ） in a dose-dependent manner. And the 20μg/ml silibinin group had the most prominent effect. Conclusion： Silibinin could suppress the expressions of collagen Ⅲ and CTGF induced by TGF-β1. It is suggested that silibinin might exert an inhibitive effect on renal fibrosis,which may be related with its role in reducing the expression of CTGF and preventing accumulation of ECM.