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目的探讨SH2B3在心肌纤维化过程中的表达调控机制。方法体外培养心肌细胞,在接受不同浓度转化生长因子-β1(10、50、100 ng/m L)和不同缺氧时间(3、6、12、24、48 h)处理后,采用Western blotting检测SH2B3蛋白的表达;采用qRT-PCR法检测SH2B3 mRNA和miR-34a的表达;在心肌细胞中转染miR-34a类似物和miR-34a抑制剂后,采用Western blotting法检测SH2B3蛋白的表达;采用荧光素酶报告基因法检测miR-34a对SH2B3基因3’UTR区活性的影响。结果在不同浓度转化生长因子-β1和不同缺氧时间刺激心肌细胞后,SH2B3在mRNA和蛋白的表达水平均降低(P<0.05);miR-34a的表达水平则升高(P<0.05),miR-34a可调控SH2B3的表达(P<0.05);miR-34a通过影响SH2B3 3’UTR活性,调控其表达(P<0.05)。结论 SH2B3在参与成纤维细胞的纤维化过程中受miR-34a的表达调控。
Objective To investigate the expression and regulation of SH2B3 during myocardial fibrosis. Methods Cardiomyocytes were cultured in vitro and treated with different concentrations of TGF-β1 (10, 50, 100 ng / mL) and hypoxia (3, 6, 12, 24, 48 h) The expression of SH2B3 mRNA and miR-34a was detected by qRT-PCR. The expression of SH2B3 protein was detected by Western blotting after transfection of miR-34a analogues and miR-34a inhibitor in cardiomyocytes. Effect of miR-34a on the activity of 3’UTR of SH2B3 gene by luciferase reporter assay. Results The expression of SH2B3 at mRNA and protein levels was decreased (P <0.05) and the expression of miR-34a at different concentrations of TGF-β1 and hypoxia-stimulated cardiomyocytes (P <0.05) miR-34a regulated the expression of SH2B3 (P <0.05); miR-34a regulated SH2B3 3’UTR activity and regulated its expression (P <0.05). Conclusion SH2B3 is regulated by the expression of miR-34a during fibroblast fibrosis.