罗沙司他通过抑制凋亡和炎症反应改善小鼠心肌缺血再灌注损伤

doi:10.11958/20240445

天津医药 ›› 2024, Vol. 52 ›› Issue (11): 1146-1151.doi: 10.11958/20240445

罗沙司他通过抑制凋亡和炎症反应改善小鼠心肌缺血再灌注损伤

蔡灯塔¹(), 常静宜², 贾姗姗², 屠荫琼³^,^△()

1 河南大学第一附属医院药学部（邮编450052）
2 河南大学抗体药物开发技术国家地方联合工程实验室
3 河南大学基础医学院医用生物实验室

收稿日期:2024-04-29 修回日期:2024-07-19 出版日期:2024-11-15 发布日期:2024-11-12
通讯作者: △E-mail：30070060@vip.henu.edu.cn
作者简介:蔡灯塔（1977），女，副主任药师，主要从事医院药事管理、临床药学方面研究。E-mail：caidengta@126.com
基金资助:
河南省医学科技攻关计划联合共建项目(LHGJ20230412)

Roxadustat improves myocardial ischemia-reperfusion injury in mice by inhibiting apoptosis and inflammatory response

CAI Dengta¹(), CHANG Jingyi², JIA Shanshan², TU Yinqiong³^,^△()

1 Department of Pharmacy, the First Affiliated Hospital of Henan University, Zhengzhou 450052, China
2 Henan University Joint National Laboratory for Antibody Drug Engineering
3 Laboratory of Medical Biology, School of Basic Medical Science, Henan University

Received:2024-04-29 Revised:2024-07-19 Published:2024-11-15 Online:2024-11-12
Contact: △E-mail：30070060@vip.henu.edu.cn

蔡灯塔, 常静宜, 贾姗姗, 屠荫琼. 罗沙司他通过抑制凋亡和炎症反应改善小鼠心肌缺血再灌注损伤[J]. 天津医药, 2024, 52(11): 1146-1151.

CAI Dengta, CHANG Jingyi, JIA Shanshan, TU Yinqiong. Roxadustat improves myocardial ischemia-reperfusion injury in mice by inhibiting apoptosis and inflammatory response[J]. Tianjin Medical Journal, 2024, 52(11): 1146-1151.

摘要/Abstract

摘要：

目的探讨罗沙司他对小鼠心肌缺血-再灌注（I/R）损伤的改善作用及相关机制。方法 24只雄性C57BL/6N小鼠随机分为假手术组、对照组和罗沙司他组，每组8只。对照组灌胃含5%二甲基亚砜的生理盐水100 μL，罗沙司他组灌胃25 mg/kg罗沙司他100 μL，将2组小鼠左冠状动脉前降支结扎40 min，再灌注24 h，建立小鼠心肌I/R模型；假手术组仅对冠状动脉左前穿针，不结扎。氯化三苯基四氮唑（TTC）染色检测小鼠心肌梗死面积，原位末端标记法（TUNEL）染色检测小鼠心肌细胞的凋亡情况，免疫组化染色检测凋亡相关蛋白bcl-2相关X蛋白质（Bax）、胱天蛋白酶3（Caspase3）表达及炎性细胞标志物F4/80、髓过氧化物酶（MPO）浸润情况，苏木精-伊红（HE）染色观察小鼠心肌细胞损伤情况。结果罗沙司他组小鼠心肌梗死面积小于假手术组和对照组（P<0.05）。对照组和罗沙司他组细胞凋亡数量多于假手术组，罗沙司他组细胞凋亡数量少于对照组（P<0.05）。对照组和罗沙司他组心肌组织中Bax、Caspase3蛋白表达水平高于假手术组，罗沙司他组低于对照组（P<0.05）。罗沙司他组心肌组织中F4/80和MPO蛋白表达水平低于对照组（P<0.05）。对照组小鼠心肌组织排列紊乱、组织间隙空泡增多；与对照组相比，罗沙司他组心肌细胞排列较整齐，组织间隙空泡减少。结论罗沙司他可缩小I/R损伤后心肌梗死面积，抑制心肌细胞凋亡，减轻心肌损伤，减少心肌巨噬细胞和中性粒细胞浸润，降低炎症损伤。

关键词: 心肌再灌注损伤, 细胞凋亡, 炎症, 罗沙司他

Abstract:

Objective To investigate the improvement effect and related mechanism of roxadustat on myocardial ischemia-reperfusion (I/R) injury in mice. Methods Twenty four male C57BL/6N mice were randomly divided into the sham operation group, the control group and the roxadustat group, with eight mice in each group. A mouse myocardial I/R model was established. The control group was given 100 μL saline injection containing 5% dimethyl sulfoxide by gavage. The roxadustat group was given 25 mg/kg roxadustat by gavage. The left anterior descending coronary artery of mice in both groups was ligated for 40 minutes, and then reperfusion for 24 hours to establish the myocardial I/R model. In the sham operation group, only the left anterior coronary artery was pierced without ligation. The area of myocardial infarction in mice was detected by triphenyltetrazolium chloride (TTC) staining. The apoptosis of mouse cardiomyocytes was detected by TdT-mediated dUTP nick and labeling (TUNEL) staining. The expression of apoptosis-related proteins bcl-2 associated X protein (Bax), Caspase3 and inflammatory cell markers F4/80 and myeloperoxidase (MPO) were detected by immunohistochemistry staining. The damage of myocardial cells was observed by hematoxylin-eosin (HE) staining. Results The area of myocardial infarction after myocardial I/R was reduced in the roxadustat group compared to the control group and the sham operation group (P<0.05). The number of apoptotic cells was higher in the control group and the roxadustat group than that in the sham operation group, and the number of apoptotic cells was lower in the roxadustat group than that in the control group (P<0.05). The expression levels of Bax and Caspase3 proteins in myocardial tissue were higher in the control group and the roxadustat group than those in the sham operation group, while those of the roxadustat group was lower than those of the control group (P<0.05). The expression levels of F4/80 and MPO proteins in myocardial tissue were lower in the roxadustat group than those in the control group (P<0.05). In the control group, the myocardial tissue arrangement was disordered, and there was an increase in interstitial vacuoles. Compared with the control group, the myocardial cells were arranged more neatly in the roxadustat group, and the interstitial vacuoles were reduced. Conclusion Roxadustat can reduce the myocardial infarction area after I/R injury, inhibit myocardial cell apoptosis, alleviate myocardial injury, reduce infiltration of myocardial macrophages and neutrophils, and reduce inflammatory injury.

Key words: myocardial reperfusion injury, apoptosis, inflammation, Roxadustat

中图分类号:

R541

图/表 6

图1 罗沙司他对小鼠心肌I/R后心肌梗死面积的影响

Fig.1 Effect of roxadustat on myocardial infarct size after I/R in mice

图2 罗沙司他对心肌I/R小鼠心肌细胞凋亡的影响（TUNEL染色，×200）

Fig.2 Effect of roxadustat on myocardial cell apoptosis in I/R mice (TUNEL staining， ×200)

表1 各组小鼠心肌I/R后心肌凋亡蛋白表达的比较（n=4，$\bar{x} \pm s$）

Tab.1 Comparison of gene expression of myocardial apoptosis in myocardial I/R mice between the three groups

组别	Bax	Caspase3
假手术组	0.33±0.17	0.16±0.09
对照组	7.16±0.98^a	2.41±0.20^a
罗沙司他组	1.39±0.36^ab	0.86±0.35^ab
F	108.000^＊	71.280^＊

图3 罗沙司他对小鼠I/R后心肌凋亡蛋白表达的影响（免疫组化染色，×200）

Fig.3 Effect of roxadustat on myocardial apoptosis gene expression after myocardial I/R in mice (immunolhistochemistry staining, ×200)

表2 各组小鼠心肌I/R后心肌巨噬细胞浸润情况比较（n=4，$\bar{x} \pm s$）

Tab.2 Comparison of myocardial macrophage infiltration after myocardial I/R between three groups of mice

组别	F4/80	MPO
假手术组	1.01±0.64	1.89±0.21
对照组	4.34±0.81^a	5.10±2.15^a
罗沙司他组	0.20±0.06^b	1.48±0.30^b
F	40.470^＊	16.370^＊

图4 罗沙司他对小鼠心肌I/R后心肌损伤的影响（×200）

Fig.4 Effect of roxadustat on myocardial injury after myocardial I/R in mice (×200)

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罗沙司他通过抑制凋亡和炎症反应改善小鼠心肌缺血再灌注损伤

Roxadustat improves myocardial ischemia-reperfusion injury in mice by inhibiting apoptosis and inflammatory response

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