RREB1对TGF⁃β1诱导系膜细胞增殖的影响
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国家自然科学基金(81970664);江苏省自然科学基金(BK20191082);江苏省妇幼健康科研项目(F201855);“789”卓越人才培养计划(789ZYRC202080119,789ZYRC202090251)


Effects of RREB1 on mesangial cell proliferation induced by TGF⁃β1
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    摘要:

    目的:探讨ras反应元件结合蛋白1(ras responsive element binding protein 1, RREB1)对转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导大鼠系膜细胞增殖的影响。方法:用TGF-β1及RREB1-siRNA转染干预系膜细胞,CCK-8法检测系膜细胞增殖活力,RT-PCR法检测细胞增殖核抗原(proliferating cell nuclear antigen, PCNA)、平滑肌肌动蛋白α(α-smooth muscle actin,α-SMA)及RREB1 mRNA水平,Western blot法检测PCNA、α-SMA及RREB1蛋白的表达,流式细胞术检测系膜细胞周期。结果:与对照组相比较,TGF-β1干预后CCK-8法检测系膜细胞增殖活力增加(P < 0.01),PCNA、α-SMA及RREB1 mRNA和蛋白水平增加(P < 0.01),细胞周期检测提示S期细胞百分率增多,G1期细胞百分率减少(P < 0.01);与TGF-β1组相比较,RREB1-siRNA转染+TGF-β1干预后的系膜细胞增殖活力下降(P < 0.01),PCNA、α-SMA及RREB1 mRNA和蛋白水平下降(P < 0.01),细胞周期提示S期细胞百分率降低,G1期细胞百分率升高(P < 0.01)。结论:TGF-β1通过上调RREB1的表达诱导系膜细胞增殖。

    Abstract:

    Objective:To investigate the effects of ras responsive element binding protein 1(RREB1) on mesangial cell proliferation induced by transforming growth factor-β1(TGF-β1). Methods:Mesangial cells were cultured in vitro,and transfected with TGF-β1 and RREB1-siRNA. The proliferation activity of mesangial cells were detected by CCK-8. Proliferating cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA) and RREB1 mRNA levels were detected by RT-PCR. The expression levels of PCNA,α-SMA and RREB1 proteins were detected by Western blot. Cell cycles of mesangial cells were detected by flow cytometry. Results: Compared with the control group, proliferation activity of mesangial cells in the TGF-β1 group was significantly increased(P < 0.01). The mRNA and protein levels of PCNA,α-SMA and RREB1 levels in cells were significantly increased(P < 0.01). The cell cycle indicated that the percentage of S phase cells increased and the percentage of G1 phase cells decreased(P < 0.01). In RREB1-siRNA+TGF-β1 group,proliferation activity of mesangial cells was significantly decreased compared with TGF-β1 group(P < 0.01). The expressions of PCNA,α-SMA and RREB1 levels in cells were significantly decreased(P < 0.01). The cell cycle indicated that the percentage of S phase cells decreased and the percentage of G1 phase cells increased(P < 0.01). Conclusion:TGF-β1 induces mesangial cells proliferation by up-regulating RREB1 expression.

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赵雅洁,施会敏,何田田,季嘉玲,甘卫华,张爱青. RREB1对TGF⁃β1诱导系膜细胞增殖的影响[J].南京医科大学学报(自然科学版),2022,42(1):8-13

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  • 收稿日期:2021-02-05
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  • 在线发布日期: 2022-01-27
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