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黄芪甲苷Ⅳ调控Nrf2/Bach1/HO-1信号通路并抑制缺氧复氧诱导的H9c2心肌细胞损伤

[Astragaloside Ⅳ regulates Nrf2/Bach1/HO-1 signaling pathway and inhibits H9c2 cardiomyocyte injury induced by hypoxia-reoxygenation].

作者信息

Yang Ping, Zhou Yu-Ping, Chang Xiu-Chun, Wang Feng, Li Gao-Wen

机构信息

Ningbo College of Health Sciences Ningbo 315100, China.

the Affiliated Hospital of Medical School of Ningbo University Ningbo 315020, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Jun;44(11):2331-2337. doi: 10.19540/j.cnki.cjcmm.20190312.001.

Abstract

Astragaloside Ⅳ(AS-Ⅳ) has protective effects against ischemia-reperfusion injury(IRI), but its mechanism of action has not yet been determined. This study aims to investigate the protective effects and mechanism of AS-Ⅳ on H9c2 cardiomyocyte injury induced by hypoxia-reoxygenation(H/R). The H/R model of myocardial cells was established by hypoxic culture for 12 hours and then reoxygenation culture for 8 hours. After AS-Ⅳ treatment, cell viability, the reactive oxygen species(ROS) levels, as well as the content or activity of superoxide dismutase(SOD), malondialdehyde(MDA), interleukin 6(IL-6), and tumor necrosis factor alpha(TNF-α), were measured to evaluate the effect of AS-Ⅳ treatment. The effect of AS-Ⅳ on HO-1 protein expression and nuclear Nrf2 and Bach1 protein expression was determined by Western blot. Finally, siRNA was used to knock down HO-1 gene expression to observe its reversal effect on AS-Ⅳ intervention. The results showed that as compared with the H/R model group, the cell viability was significantly increased(P<0.01), ROS level in the cells, MDA, hs-CRP and TNF-α in cell supernatant and nuclear protein Bach1 expression in the cells were significantly decreased(P<0.01), while SOD content, HO-1 protein expression in cells and expression of nuclear protein Nrf2 were significantly increased(P<0.01) in H/R+AS-Ⅳ group. However, pre-transfection of HO-1 siRNA into H9c2 cells by liposome could partly reverse the above effects of AS-Ⅳ after knocking down the expression of HO-1. This study suggests that AS-Ⅳ has significant protective effect on H/R injury of H9c2 cardiomyocytes, and Nrf2/Bach1/HO-1 signaling pathway may be a key signaling pathway for the effect.

摘要

黄芪甲苷Ⅳ(AS-Ⅳ)对缺血再灌注损伤(IRI)具有保护作用,但其作用机制尚未明确。本研究旨在探讨AS-Ⅳ对缺氧复氧(H/R)诱导的H9c2心肌细胞损伤的保护作用及机制。通过缺氧培养12小时,然后复氧培养8小时建立心肌细胞H/R模型。AS-Ⅳ处理后,检测细胞活力、活性氧(ROS)水平以及超氧化物歧化酶(SOD)、丙二醛(MDA)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的含量或活性,以评估AS-Ⅳ处理的效果。通过蛋白质免疫印迹法检测AS-Ⅳ对HO-1蛋白表达以及核Nrf2和Bach1蛋白表达的影响。最后,使用小干扰RNA(siRNA)敲低HO-1基因表达,以观察其对AS-Ⅳ干预的逆转作用。结果显示,与H/R模型组相比,H/R+AS-Ⅳ组细胞活力显著升高(P<0.01),细胞内ROS水平、细胞上清液中MDA、超敏C反应蛋白(hs-CRP)和TNF-α以及细胞内核蛋白Bach1表达显著降低(P<0.01),而SOD含量、细胞内HO-1蛋白表达以及核蛋白Nrf2表达显著升高(P<0.01)。然而,通过脂质体将HO-1 siRNA预先转染至H9c2细胞中,在敲低HO-1表达后可部分逆转AS-Ⅳ的上述作用。本研究表明,AS-Ⅳ对H9c2心肌细胞的H/R损伤具有显著保护作用,Nrf2/Bach1/HO-1信号通路可能是其发挥作用的关键信号通路。

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