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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 secretion of type III collagen in human proximal tubular epithelial cells (HK-2) induced by transforming growth factor (TGF)-β_1. Methods: The cultured HK-2 cells were divided into 3 groups: 1 control group; 2 TGF-β 1 group (final concentration of TGF-β 1 was 5 ng/ml); 3 TGF-ββ+ silybin group (TGF-β 1 The final concentration was 5 ng/ml, and the final silybin concentrations were 5, 10, and 20 μg/ml, respectively. After 48 h, the level of CTGF mRNA in cells was measured by reverse transcription polymerase chain reaction (RT-PCR), and the concentrations of collagen III and CTGF in culture supernatants were measured by ELISA. RESULTS: Compared with the control group, TGF-β_1 stimulation significantly increased CTGF gene expression and protein secretion and type III collagen secretion in HK-2 cells (P<0.01). Compared with TGF-β_1 group, addition of 5-20 μg/ml silibinin inhibited CTGF gene expression and protein secretion and type III collagen secretion induced by TGF-β_1 stimulation (P<0.01), and it was treated with 20 μg/ml water. Outstanding effects are most noticeable. CONCLUSION: Silybin can partially inhibit the expression of TGF-β 1 -induced CTGF gene and protein, and the secretion of type III collagen. It suggests that silibinin may have potential anti-renal effect by inhibiting the expression of CTGF and reducing the accumulation of extracellular matrix. Fibrosis.