Activation of FXR receptor reduces damage of ET-1 on H9C2 cardiomyocytes by activating AMPK signaling pathway.

IF 4.3 4区医学 0 MEDICINE, GENERAL & INTERNAL

Panminerva medica Pub Date : 2024-06-01 Epub Date: 2020-05-14 DOI:10.23736/S0031-0808.20.03930-0

Xiaoli Li, Zhen Zeng, Jiuchang Zhong, Hongjiang Wang, Xinchun Yang

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引用次数: 0

Abstract

Background: The aim of this study was to investigate the effect of Farnesoid X receptor (FXR) on oxidative stress injury of H9C2 cardiomyocytes induced by endothelin-1 (ET-1), and to explore the possible mechanism.

Methods: H9C2 cardiomyocytes were treated with ET-1 at concentrations of 10-8, 10-7 and 10-6 mmol/L for 12, 24, 36, and 48 h, respectively. At the same time, oxidative stress injury models were established. After the oxidative stress injury model was established, GW4064 (FXR agonist), siRNA-FXR (the virus interfered with FXR expression) and FXR empty virus were treated for 48 h. CCK-8 method was used to observe the survival rate of cardiomyocytes. Biochemical kit method was used to detect Creatine Kinase (CK) contents, CAT, and mitochondrial activity. Western blot was used to detect the protein expression of SOD1, AMPK and p-AMPK. And real-time PCR was used to detect GPX1 mRNA, GPX3 mRNA, Sirt1 mRNA, PGC-1α mRNA expression levels.

Results: At the concentrations of 10-8, 10-7 and 10-6 mmol/L, ET-1 could induce the decrease of mitochondrial complex I and III activity and increase of CK content in H9C2 cardiomyocytes. Moreover, with the increase of ET-1 concentration and the extension of culture time, the oxidative stress damage of cardiomyocytes became more serious. After ET-1 intervention, the expressions of p-AMPK protein, Sirt1 mRNA and PGC-1α mRNA were decreased. The 2 μmol/L GW4064 could effectively improve the oxidative stress induced by ET-1, and compared with the ET-1 group, the survival rate of H9C2 cardiomyocytes increased obviously. In addition, the CK content was decreased. On the contrary, the mitochondrial complex I and III, and CAT activity increased significantly, and the expressions of SOD1 and p-AMPK protein, Sirt1 mRNA and PGC-1α mRNA increased obviously. However, siRNA-FXR can partially block the improvement effect of FXR agonist on cardiomyocytes injury.

Conclusions: Activation of FXR receptor may reduce the damage of ET-1 on H9C2 cardiomyocytes by activating AMPK signaling pathway.

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激活 FXR 受体可通过激活 AMPK 信号通路减轻 ET-1 对 H9C2 心肌细胞的损伤。

背景：方法：分别用10-8、10-7和10-6 mmol/L浓度的ET-1处理H9C2心肌细胞12、24、36和48 h。同时建立氧化应激损伤模型。氧化应激损伤模型建立后，分别用 GW4064（FXR 激动剂）、siRNA-FXR（干扰 FXR 表达的病毒）和 FXR 空病毒处理 48 小时，用 CCK-8 法观察心肌细胞的存活率。生化试剂盒法检测肌酸激酶（CK）含量、CAT和线粒体活性。用 Western 印迹法检测 SOD1、AMPK 和 p-AMPK 的蛋白表达。实时 PCR 检测 GPX1 mRNA、GPX3 mRNA、Sirt1 mRNA、PGC-1α mRNA 的表达水平：结果：在 10-8、10-7 和 10-6 mmol/L 的浓度下，ET-1 能诱导 H9C2 心肌细胞线粒体复合物 I 和 III 活性的降低和 CK 含量的增加。此外，随着 ET-1 浓度的增加和培养时间的延长，心肌细胞的氧化应激损伤更加严重。ET-1干预后，p-AMPK蛋白、Sirt1 mRNA和PGC-1α mRNA的表达均下降。2 μmol/L GW4064能有效改善ET-1诱导的氧化应激，与ET-1组相比，H9C2心肌细胞的存活率明显提高。此外，CK 含量也有所下降。相反，线粒体复合物 I、III 和 CAT 活性明显增加，SOD1 和 p-AMPK 蛋白、Sirt1 mRNA 和 PGC-1α mRNA 表达明显增加。然而，siRNA-FXR 能部分阻断 FXR 激动剂对心肌细胞损伤的改善作用：结论：激活 FXR 受体可通过激活 AMPK 信号通路减轻 ET-1 对 H9C2 心肌细胞的损伤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Panminerva medica 医学-医学：内科

CiteScore

5.00

自引率

2.30%

发文量

199

审稿时长

>12 weeks

期刊介绍： Panminerva Medica publishes scientific papers on internal medicine. Manuscripts may be submitted in the form of editorials, original articles, review articles, case reports, special articles, letters to the Editor and guidelines. The journal aims to provide its readers with papers of the highest quality and impact through a process of careful peer review and editorial work. Duties and responsibilities of all the subjects involved in the editorial process are summarized at Publication ethics. Manuscripts are expected to comply with the instructions to authors which conform to the Uniform Requirements for Manuscripts Submitted to Biomedical Editors by the International Committee of Medical Journal Editors (ICMJE).