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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年05期

页码:

410-414,444

栏目:

基础医学

出版日期:

2024-09-25

文章信息/Info

Title:

The protective effect of liraglutide on palmitic acid induced myocardial cell injury

文章编号:

1006-8147（2024）05-0410-06

作者:

张子钊; 段军滢; 高怡; 王凯; 张帆; 张跃; 李广平

（天津市心血管病离子与分子机能重点实验室，天津医科大学第二医院心脏科，天津心脏病学研究所，天津 300211）

Author(s):

ZHANG Zizhao; DUAN Junying; GAO Yi; WANG Kai; ZHANG Fan; ZHANG Yue; Li Guangping

（Tianjin Key Laboratory of Ionic-Molecular Function of Cardiovascular Disease，Department of Cardiology，Tianjin Institute of Cardiology，the Second Hospital of Tianjin Medical University，Tianjin 300211，China）

关键词:

利拉鲁肽; H9C2细胞; 脂毒性; 心肌保护; 凋亡蛋白

Keywords:

liraglutide; H9C2 cells; lipotoxicity; cardioprotection; apoptotic protein

分类号:

R541.9

DOI:

10.20135/j.issn.1006-8147.2024.05.0410

文献标志码:

A

摘要:

目的：探讨利拉鲁肽（lira）对棕榈酸（PA）所致 H9C2 大鼠心肌细胞脂毒性损伤的干预效果及其相关机制。方法：在不同浓度的PA（0、50、100、150、200、300 μmol/L）和不同时间（6、12、24 h）的条件下分别刺激 H9C2大鼠心肌细胞（n=3），建立心肌细胞的脂毒性损伤模型，使用CCK8检测确定使H9C2 细胞脂毒性损伤较大、细胞存活数最多的刺激条件。将成功建立模型的H9C2细胞分为对照组（CON）、CON＋Lira（1 000 nmol/L）组、CON＋PA（150 μmol/L）组和PA（150 μmol/L）＋Lira（1 000 nmol/L）组进行油红O染色（n=3），检测细胞内脂质含量，以明确PA（150 μmol/L）对心肌细胞有无脂毒性。将H9C2细胞根据不同的PA浓度（0、50、100、150、200、300 μmol/L）分为6个组（CON、PA1、PA2、PA3、PA4和PA5）（n=3），按照不同的Lira浓度分为6组：CON组、PA组（150 μmol/L）、CON+Lira3（1 000 nmol/L）组、PA（150 μmol/L）+Lira1（100 nmol/L）组、PA（150 μmol/L）+Lira2（500 nmol/L）组和PA（150 μmol/L）+Lira3（1 000 nmol/L）组（n=3），用蛋白印迹测定凋亡相关蛋白的表达量及其与PA、Lira浓度的关系。结果：使用CCK8检测并最终确定PA为150 μmol/L 、刺激 12 h 的条件下，H9C2 细胞的脂毒性损伤较大、细胞存活数最多。与CON组相比，PA组（150 μmol/L）的细胞活性明显下降、脂滴明显增加（t=34.53，P＜0.05），经lira 预处理后脂滴明显减少（t=19.07，P＜0.05）；与CON组相比，随着PA浓度的增加，Bax、Caspase-3 蛋白表达明显增加（F=12.32、3.307，均P＜0.05），Bcl-2 蛋白表达明显减少（F=7.618，P＜0.05）；与 PA 刺激组（150 μmol/L）相比，lira预处理可以使 Bcl-2 蛋白表达明显增加（F=7.104，P＜0.05，），Bax、Caspase-3 蛋白表达明显减少（F=12.32、7.104，均P＜0.05）。结论：浓度为 150 μmol/L 的 PA 刺激H9C2心肌细胞 12 h 后可造成心肌细胞脂毒性损伤模型，lira 可以改善甚至逆转PA诱导的 H9C2 心肌细胞脂毒性损伤，其机制可能与抑制炎症、抑制细胞凋亡有关。

Abstract:

Objective：To explore the protective effect of liraglutide（lira）on palmitic acid（PA）induced H9C2 rat cardiomyocyte lipotoxic injury model and its possible mechanism. Methods：First，H9C2 cells were stimulated with different concentrations of PA （0，50，100，150，200，300 μmol/L）and different times（6，12，24 hours）to establish a model of lipotoxic injury in cardiomyocytes（n=3）. CCK8 was used to determine the stimulus conditions that resulted in greater lipotoxic damage to H9C2 cells and the highest number of viable cells.The H9C2 cells that had successfully established the model were divided into control group（CON），CON+Lira（1 000 nmol/L） group，CON+PA（150 μmol/L）group，and PA（150 μmol/L）+Lira（1 000 nmol/L） group.Oil red O staining was used to detect the intracellular lipid content to determine whether PA（150 μmol/L） was lipid toxic to cardiomyocytes（n=3）.According to different PA concentrations，the H9C2 cells was divided into 6 groups（CON，PA1，PA2，PA3，PA4 and PA5）（n=3），and different lira concentrations were divided into CON group，PA，CON+Lira3（1 000 nmol/L）group，PA+Lira1（100 nmol/L） group，PA+Lira2（500 nmol/L） group and PA+Lira3（1 000 nmol/ L） group（n=3）.The expression of apoptosis-related proteins and their relationship with PA and lira concentrations were determined by Western blotting（n=3）. Results：CCK8 was used to detect and finally determine that 150 μmol/L PA and stimulated for 12 h，the lipotoxic damage of H9C2 cells was greater and the number of cell survival was the highest.Compared with CON group，the cell activity decreased and lipid droplets increased significantly in the PA stimulation group（150 μmol/L）（t=34.53，P＜0.05），and the lipid droplets decreased after lira pretreatment（t=19.07，P＜0.05）. Compared with the CON group，with the increase of PA concentration，the expression of Bax and Caspase-3 proteins increased significantly（F=12.32，3.307，both P＜0.05），and the expression of Bcl-2 protein decreased significantly（F=7.618，P＜0.05）.Compared with the PA stimulation group，the expression of Bcl-2 protein was significantly increased by lira pretreatment（F=7.104，P<0.05），and the expression of Bax and Caspase-3 proteins was significantly decreased（F=12.32，7.104，both P<0.05）. Conclusion：Stimulating H9C2 cardiomyocytes with a concentration of 150 ?滋mol/L PA for 12 h can induce a model of myocardial cell lipotoxic damage. Lira can improve or even reverse the lipotoxic damage in H9C2 cardiomyocytes induced by PA，which may be related to the inhibition of inflammation and apoptosis.

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备注/Memo

备注/Memo:

基金项目 国家自然科学基金面上项目 （ 82270336 ）
作者简介 张子钊（1998-），男，医师，硕士，研究方向：冠心病介入治疗和心力衰竭的诊治；通信作者：李广平，E-mail：tic_tjcardiol@126.com。

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更新日期/Last Update: 2024-09-20