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人参皂苷Rb1通过调节TGF-β 1/Smad3信号通路抑制肾小管上皮

        细胞-间质转化                Δ


        刘志文 ,徐江雁 ,张振强 ,张效威 ,高 改 ,王萌萌 ,王 辉 ,李真珍 ,谢治深 (1.河南中医药大学中医药
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        科学院,郑州 450046;2.河南中医药大学药学院,郑州 450046;3.郑州大学第一附属医院医学研究中心,郑州
        450052)
        中图分类号 R965          文献标志码 A          文章编号 1001-0408(2022)05-0535-07
        DOI  10.6039/j.issn.1001-0408.2022.05.05
        摘 要 目的 研究人参皂苷Rb1(G-Rb1)对肾小管上皮细胞-间质转化(EMT)的影响及可能机制。方法 采用10 ng/mL 生长因子β 1
       (TGF-β 1 )诱导人肾小管上皮细胞 HK-2 发生 EMT。观察 10、20、30 μmol/L G-Rb1 作用 48 h 后 HK-2 细胞的形态学变化;测定 1.0、
        2.5、5.0、10、20、30 μmol/L G-Rb1 作用24 h后人胚胎肾细胞HEK293中报告基因载体SBE转录活性,并测定上述浓度G-Rb1 作用
        24 h后对HK-2细胞相对活力的影响。检测10、20、30 μmol/L G-Rb1作用24 h后HK-2细胞中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶
        原蛋白(COL-Ⅰ)、纤连蛋白(FN)mRNA的表达;检测10、20、30 μmol/L G-Rb1作用24 h后HK-2细胞中α-SMA、Smad家族成员3
       (Smad3)、磷酸化Smad3(p-Smad3)、COL-Ⅰ、FN和E-钙黏蛋白(E-cadherin)的表达。结果 10~30 μmol/L G-Rb1能够抑制TGF-β 1
        诱导的HK-2细胞发生EMT,显著抑制因TGF-β1刺激导致的SBE转录活性升高(P<0.05),并且对HK-2细胞的相对活力无明显影
        响(P>0.05)。在 TGF-β 1诱导下,细胞中α-SMA、COL-Ⅰ、FN 蛋白及其 mRNA 和 Smad3、p-Smad3 蛋白的表达均显著上调(P<
        0.05),E-cadherin蛋白的表达显著下调(P<0.05);而G-Rb1则可有效地逆转上述蛋白或mRNA的表达。结论 G-Rb1可在一定程
        度上保护肾小管上皮细胞免受TGF-β1诱导引起的EMT,这可能与其抑制了TGF-β1/Smad3信号通路的激活有关。
        关键词 人参皂苷Rb1;TGF-β1/Smad3信号通路;转化生长因子β1;肾小管上皮细胞;上皮细胞-间质转化

        Inhibition of ginsenoside Rb1 on the epithelial-mesenchymal transition of renal tubular epithelial cells by
        regulating TGF-β1/Smad3 signaling pathway
        LIU Zhiwen ,XU Jiangyan ,ZHANG Zhenqiang ,ZHANG Xiaowei ,GAO Gai ,WANG Mengmeng ,WANG
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        Hui ,LI Zhenzhen ,XIE Zhishen (1. Academy of Chinese Medical Science,Henan University of Chinese
        Medicine,Zhengzhou 450046,China;2. College of Pharmacy,Henan University of Chinese Medicine,
        Zhengzhou 450046,China;3. Medical Research Center,the First Affiliated Hospital of Zhengzhou University,
        Zhengzhou 450052,China)
        ABSTRACT    OBJECTIVE To study the effects of ginsenoside Rb1 (G-Rb1) on epithelial-mesenchymal transition(EMT) of
        renal tubular epithelial cells and its potential mechanism. METHODS The growth factor β 1(TGF-β 1 )10 ng/mL was used to induce
        EMT of human renal tubular epithelial cells HK-2. The morphological changes of HK-2 cells were observed after treated with 10,
        20,30 μmol/L G-Rb1 for 48 h. The transcriptional activities of biovector SBE in human embryonic kidney cell HEK293 were
        determined after 24 h treatment with 1.0,2.5,5.0,10,20,30 μmol/L G-Rb1. Effects of above concentration of G-Rb1 on the
        viability of HK-2 cells were determined after 24 h of treatment. mRNA expressions of α-smooth muscle actin(α-SMA),collagen Ⅰ
       (COL-Ⅰ)and fibronectin(FN)in HK-2 cells were detected after treated with 10,20,30 μmol/L G-Rb1 for 24 h. The expressions
        of α-SMA,Smad3,p-Smad3,COL-Ⅰ,FN and E-cadherin were detected after treated with 10,20,30 μmol/L G-Rb1 for 24 h.
        RESULTS G-Rb1 of 10-30 μmol/L significantly inhibited TGF-β 1-induced EMT in HK-2 cells and the increase of transcriptional
        activities of biovector SBE induced by TGF-β 1(P<0.05),but had no effects on relative activities of HK-2 cells(P>0.05). The
                                                           protein and mRNA expressions of α-SMA,COL-Ⅰ and FN,
           Δ 基金项目:国家重点研发计划“战略性科技创新合作”重点专项                  the protein expressions of Smad3 and p-Smad3 were
       (No.2020YFE0201800);河南省高校科技创新团队支持计划项目
                                                           significantly up-regulated induced by TGF-β 1(P<0.05),while
       (No.21IRTSTHN026);河南省高等学校青年骨干教师培养计划项目
                                                           the protein expression of E-cadherin was significantly down-
       (No.2021GGJS081)
                                                           regulated (P<0.05);G-Rb1 could effectively reverse above
           *硕士研究生。研究方向:中药药效物质基础与作用机制。电
                                                           protein or mRNA expressions. CONCLUSIONS G-Rb1 can
        话:0371-65680206。E-mail:1649664828@qq.com
           # 通信作者:副教授,硕士生导师,博士。研究方向:中药药效物                  protect renal tubular epithelial cells from EMT induced by
        质 基 础 与 作 用 机 制 。 电 话 :0371-65680206。 E-mail:xiezhishen  TGF-β 1 to a certain extent,which may be related to inhibiting
        @hactcm.edu.cn                                     the activation of TGF-β1/Smad3 signaling pathway.


        中国药房    2022年第33卷第5期                                               China Pharmacy 2022 Vol. 33 No. 5  ·535 ·
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