Basigin/CD147参与马兜铃酸损伤人肾小管上皮细胞的体外实验

目的:探讨马兜铃酸(AA)对人肾小管上皮细胞损伤机制及细胞外基质金属蛋白酶诱导因子(Basigin/CD147)在损伤过程中发挥的作用。方法:不同浓度AA(10、20、40μg/ml)作用于人近端肾小管上皮细胞(HK-2)不同时间(24、48 h)后,MTT比色法检测细胞增殖变化;Basigin/CD147干扰质粒转染HK-2,选取AA最适浓度(20μg/ml),ELISA法检测不同时间段(24、48 h)细胞上清液中转化生长因子-β1(TGF-β1)含量;实时定量(Real-time)PCR法检测Basigin/CD147 mRNA表达;Western印迹检测Basigin/CD147、TGF-β1和α-平滑肌肌动蛋白(α-SMA)的蛋白表达。结果:AA 48 h组TGF-β1含量显著高于空白组(P0.05),siRNA+AA 48 h组TGF-β1低于48 h AA组(P0.05);AA48 h组Basigin/CD147 mRNA含量显著高于空白组(P0.05),siRNA+AA 48 h组Basigin/CD147 mRNA显著低于48 h AA组(P0.05);AA 48 h组Basigin/CD147、α-SMA、TGF-β1蛋白表达明显高于空白组(P0.05),siRNA+AA 48 h组Basigin/CD147、α-SMA、TGF-β1蛋白表达均显著低于48 h AA组(P0.01)。结论:Basigin/CD147参与介导AA所致HK-2细胞损伤过程,干扰Basigin/CD147表达可能抑制AA引起的细胞纤维化;Basigin/CD147蛋白表达调控异常可能是引起马兜铃酸肾病的重要发病机制之一。

Effect of Basigin /CD147 on Human Renal Tubular Epithelial Cells Injured by Aristolochic Acid in Vitro

Objective： To investigate the injury mechanism of aristolochic acid（ AA） on human renal tubular epithelial cell and the regulation effect of Basigin /CD147 during the process. Methods： The human renal proximal tubular epithelial cells（ HK-2） were treated with AA at dosages of 10,20,40 μg /ml at various times of 24 h,48 h. Then the cell proliferation was determined by MTT assay. Transfected HK-2 with Basigin /CD147 interference plasmid,ELISA was used to detect the content of transforming growth factor（ TGF）-β1in the cell supernatant when opposed to the optimal concentration of AA at20 μg / ml for 24 h,48 h. Real-time quantitative PCR was performed to evaluate the expression of Basigin /CD147 mRNA and Western blotting to Basigin /CD147,α-SMA and TGF-β1protein expressions. Results： The TGF-β1level in AA group was higher than that of the blank group significantly at 48 h（ P〈0. 05）,and the TGF-β1level in siRNA ＋ AA group was lower than that of the AA group at 48 h（ P〈0. 05）; the Basigin /CD147 mRNA in the AA group was higher than that of the blank group significantly at 48 h（ P〈0. 05）,and the Basigin /CD147 mRNA in the siRNA ＋ AA group was lower than that of the AA group at 48 h（ P〈0. 05）; Expressions of Basigin /CD147,α-SMA and TGF-β1protein in the AA group were higher than those of the blank group（ P〈0. 05）,the expressions of Basigin /CD147,α-SMA and TGF-β1protein in the siRNA ＋ AA group were lower than those of the AA group at 48 h（ P〈0. 01）. Conclusion： Basigin /CD147 involved in the process of HK-2 injury caused by AA,and interfering Basigin /CD147 expression may inhibit AA-induced cell fibrosis. The results suggest that abnormal expression of Basigin /CD147 may be one of the important pathogenesis of aristolochic acid nephropathy.