人参皂苷Rg1通过Sestrin2保护心肌细胞的作用

刘燃; 林志; 杨帆; 段继坤

doi:10.12259/j.issn.2095-610X.S20230614

人参皂苷Rg1通过Sestrin2保护心肌细胞的作用

doi: 10.12259/j.issn.2095-610X.S20230614

昆明医科大学第二附属医院心血管内科三病区，云南昆明　650031

基金项目: 云南省教育厅科学研究基金资助项目（2019J1252）

详细信息

作者简介:
刘燃（1988～），男，哈尼族，云南普洱人，医学硕士，主治医师，主要从事心血管病学临床工作

中图分类号: R541.4
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出版历程
- 收稿日期: 2023-03-16
- 网络出版日期: 2023-06-19
- 刊出日期: 2023-06-25

Mechanisms of Ginsenoside Rg1 in Protecting Cardiomyocytes by Sestrin2

Dept. of Cardiology，The second Affiliated Hospital of Kunming Medical University， Kunming Yunnan 650031，China

摘要

摘要: 目的研究人参皂苷Rg1（G-Rg1）对抗心肌缺血再灌注损伤中Sestrin2的作用及相关机制。方法体外培养大鼠 H9c2 心肌细胞，按实验要求随机分5组，分别为对照组，缺氧复氧组，G-Rg1组，缺氧诱导因子 -1α（Hypoxia Inducible Factor-1，HIF-1，HIF-1α）抑制剂组，Sestrin2 siRNA干扰组。流式细胞术检测各组心肌细胞内活性氧簇（reactive oxygen species，ROS）含量。蛋白质免疫印迹法检测HIF-1α、Sestrin2、蛋白激酶R样内质网激酶（protein kinase R-like ER kinase，PERK）、活化转录因子6（the activating transcription factor 6，ATF-6）和跨膜蛋白激酶1（inositol requiring enzyme 1，IRE-1）蛋白表达。免疫荧光法检测Sestrin2、HIF-1α、IRE-1蛋白表达。结果与对照组相比，缺氧复氧组细胞内ROS水平、HIF-1α、Sestrin2、ATF-6、PERK、IRE-1蛋白表达水平明显升高（P < 0.05）；与缺氧复氧组相比，G-Rg1组细胞内ROS水平明显降低（P < 0.05），HIF-1α、Sestrin2、ATF-6、PERK、IRE-1蛋白表达水平明显升高（P < 0.05）；与G-Rg1组比较，HIF-1α抑制剂组心肌细胞内ROS水平升高，Sestrin2蛋白表达明显下调（P < 0.05）；与G-Rg1组比较，siRNA干扰组心肌细胞内ROS水平升高，IRE-1表达明显升高（P < 0.05）。结论 G-Rg1通过上调Sestrin2表达，保护缺氧复氧心肌细胞，并维持内质网应激平衡稳态。
- 人参皂苷Rg1 /
- ATF-6 /
- PERK /
- IRE-1 /
- Sestrin2
Abstract: Objective To explore the role and mechanisms of Sestrin2 in ameliorating hypoxia-reoxygenation injury of rat’s cardiomyocytes by ginsenoside-rg1 （G-Rg1）. Methods Rat H9c2 cardiomyocytes cultured in vitro were randomly divided into 5 groups according to the experimental requirements: control group, Hypoxia reoxygenation group, G-Rg1 group, hypoxia Inducible Factor-1 （HIF-1, HIF-1α） inhibitor group, and Sestrin2 siRNA interference group. Flow cytometry was used to detect the expression levels of ROS in cardiomyocytes. Western blot was used to detect the protein expression of HIF-1α, Sestrin2, ATF-6, PERK, IRE-1 in cardiomyocytes. Immunofluorescence was used to test the protein expression of Sestrin2, HIF-1α, IRE-1. Results Compared with control group, the level of ROS and protein expression of HIF-1α、Sestrin2、ATF-6、PERK、IRE-1 of hypoxiareoxygenation group were significantly increased （P < 0.05） . Compared with the hypoxiareoxygenation group , the ROS level in G-Rg1 group were significantly reduced and protein expression of HIF-1α、Sestrin2、ATF-6、PERK、IRE-1 in G-Rg1 group were significantly increased （P < 0.05） . Compared with G-Rg1 group, the level of ROS were significantly increased and the protein level of Sestrin2 were significantly reduced, in HIF-1α inhibition group （P < 0.05）. Compared with G-Rg1 group, the level of ROS were significantly increased and the protein level of IRE-1 were significantly upregulated in Sestrin2 siRNA group （P < 0.05）. Conclusions G-Rg1 protects rat cardiomyocytes and maintains endoplasmic reticulum homeostasis in vitro by upregulating expression of Sestrin2.
- Ginsenoside-rg1 /
- ATF-6 /
- PERK /
- IRE-1 /
- Sestrin2

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图 1 各组心肌细胞 ROS 水平

用 ROS 阳性细胞所占百分比表示各组心肌细胞中 ROS 水平。^**P < 0.01。

Figure 1. The levels of ROS in cardiomyocytes

下载: 全尺寸图片幻灯片

图 2 各组心肌细胞 HIF-1α、Sestrin2 蛋白表达及内质网跨膜蛋白：ATF-6、PERK、IRE-1 表达

^*P < 0.05，^**P < 0.01。

Figure 2. The protein expression of HIF-1α，Sestrin2，ATF-6，PERK，IRE-1 in cardiomyocytes

下载: 全尺寸图片幻灯片

图 3 各组心肌细胞 HIF-1-α、Sestrin2、ATF-6、PERK、IRE-1 蛋白表达（×200）

Figure 3. The protein expression of Sestrin2，HIF-1α，ATF-6，PERK，IRE-1 in cardiomyocytes （×200）

下载: 全尺寸图片幻灯片

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