Basigin/CD147参与马兜铃酸损伤人肾小管上皮细胞的体外实验 |
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引用本文: | 池杨峰,彭文,王云满,刘育军,王利,姚卫国,王浩.Basigin/CD147参与马兜铃酸损伤人肾小管上皮细胞的体外实验[J].上海中医药大学学报,2014(4):74-78. |
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作者姓名: | 池杨峰 彭文 王云满 刘育军 王利 姚卫国 王浩 |
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作者单位: | 上海中医药大学附属普陀医院肾内科,上海200062 |
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基金项目: | 上海市教委预算内基金资助项目(2011JW61); 上海市医学重点专科建设项目(ZK2012A34) |
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摘 要: | 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。
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关 键 词: | 马兜铃酸 肾小管上皮细胞(KH-2) 细胞外基质金属蛋白酶诱导因子(Basigin CD147) 纤维化 |
Effect of Basigin /CD147 on Human Renal Tubular Epithelial Cells Injured by Aristolochic Acid in Vitro |
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Institution: | CHI Yang-feng,PENG Wen,WANG Yun-man,LIU Yu-jun ,WANG Li,YAO Wei-guo ,ANG Hao( Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine) |
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Abstract: | 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. |
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Keywords: | aristolochic acid renal tubular epithelial cell(HK-2) Basigin /CD147 fibrosis |
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