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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(P<0.05),siRNA+AA 48 h组TGF-β1低于48 h AA组(P<0.05);AA48 h组Basigin/CD147 mRNA含量显著高于空白组(P<0.05),siRNA+AA 48 h组Basigin/CD147 mRNA显著低于48 h AA组(P<0.05);AA 48 h组Basigin/CD147、α-SMA、TGF-β1蛋白表达明显高于空白组(P<0.05),siRNA+AA 48 h组Basigin/CD147、α-SMA、TGF-β1蛋白表达均显著低于48 h AA组(P<0.01)。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。
Objective: To investigate the mechanism of aristolochic acid (AA) on human renal tubular epithelial cells and the effect of extracellular matrix metalloproteinase inducer (Basigin / CD147) on the injury. Methods: The proliferation of human proximal tubular epithelial cells (HK-2) treated with different concentrations of AA (10, 20, 40μg / ml) for 24 h and 48 h were detected by MTT assay. The expressions of Basigin / CD147 HK-2 was transfected into HK-2 cells to select the optimal concentration of AA (20μg / ml). The content of transforming growth factor-β1 (TGF-β1) in the supernatant of the cells at different time points The expression of Basigin / CD147 mRNA was detected by Real-time PCR. The protein expressions of Basigin / CD147, TGF-β1 and α-smooth muscle actin (α-SMA) were detected by Western blotting. Results: The content of TGF-β1 in AA 48 h group was significantly higher than that in blank group (P <0.05), and the level of TGF-β1 in 48 h siRNA + AA group was lower than that in 48 h AA group (P <0.05) (P <0.05). The expression of Basigin / CD147 mRNA in 48 h siRNA + AA group was significantly lower than that in 48 h AA group (P <0.05) (P <0.05). The expressions of Basigin / CD147, α-SMA and TGF-β1 in siRNA + AA 48 h group were significantly lower than those in 48 h AA group (P <0.01). CONCLUSION: Basigin / CD147 is involved in mediating AA-induced injury of HK-2 cells and interfering with Basigin / CD147 expression may inhibit AA-induced cellular fibrosis. The abnormal regulation of Basigin / CD147 protein expression may be an important cause of aristolochic acid nephropathy One of the mechanisms.