人参皂苷Rb1通过调节TGF-β1/Smad3信号通路抑制肾小管上皮细胞-间质转化
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篇名: 人参皂苷Rb1通过调节TGF-β1/Smad3信号通路抑制肾小管上皮细胞-间质转化
TITLE: Inhibition of ginsenoside Rb 1 on the epithelial-mesenchymal transition of renal tubular epithelial cells by regulating TGF-β1/Smad3 signaling pathway
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10ng/mL生长因子β1(TGF-β1)诱导人肾小管上皮细胞HK-2发生EMT。观察10、20、30μmol/LG-Rb1作用48h后HK-2细胞的形态学变化;测定1.0、2.5、5.0、10、20、30μmol/LG-Rb1作用24h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24h后对HK-2细胞相对活力的影响。检测10、20、30μmol/LG-Rb1作用24h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、30μmol/LG-Rb1作用24h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10~30μmol/LG-Rb1能够抑制TGF-β1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-β1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。
ABSTRACT: OBJECTIVE To study the effects of ginsenoside Rb 1(G-Rb1)on epithelial-mesenchymal transition (EMT)of renal tubular epithelial cells and its potential mechanism. METHODS The growth factor β1(TGF-β1)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 HEK 293 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-Rb 1 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-β1-induced EMT in HK- 2 cells and the increase of transcriptional activities of biovector SBE induced by TGF-β1(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 , the protein expressions of Smad 3 and p-Smad 3 were significantly up-regulated induced by TGF-β1(P<0.05),while the protein expression of E-cadherin was significantly down- regulated(P<0.05);G-Rb1 could effectively reverse aboveprotein or mRNA expressions. CONCLUSIONS G-Rb1 can protect renal tubular epithelial cells from EMT induced byxiezhishen TGF-β1 to a certain extent ,which may be related to inhibiting the activation of TGF-β1/Smad3 signaling pathway.
期刊: 2022年第33卷第05期
作者: 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
AUTHORS: LIU Zhiwen,XU Jiangyan, ZHANG Zhenqiang,ZHANG Xiaowei,GAO Gai,WANG Mengmeng,WANG Hui,LI Zhenzhen,XIE Zhishen
关键字: 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
KEYWORDS: ginsenoside Rb 1;TGF-β1/Smad3 signaling pathway ;transforming growth factor β1;renal tubular epithelial cells ;
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