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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 ·