Role of exosomes in reduction of myocardial ischaemia-reperfusion injury by remote preconditioning of trauma in rats
10.3760/cma.j.cn131073-20240906-00909
- VernacularTitle:外泌体在远端创伤预处理减轻大鼠心肌缺血再灌注损伤中的作用
- Author:
Aoxue LI
1
;
Shuai QIN
1
;
Xinzhe WU
1
;
Min WANG
1
;
Yun WU
1
Author Information
1. 安徽医科大学第二附属医院麻醉与围术期医学科 安徽医科大学麻醉与围术期医学安徽普通高校重点实验室，合肥 203601
- Publication Type:Journal Article
- Keywords:
Exosome;
Ischemic preconditioning;
Myocardial reperfusion injury
- From:
Chinese Journal of Anesthesiology
2025;45(9):1135-1141
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the role of exosomes in reduction of myocardial ischemia-reperfusion (I/R) injury (MIRI) by remote preconditioning of trauma (RPCT) in rats.Methods:This experiment was performed in 2 parts. In vivo experiment Adult male Sprague-Dawley rats, aged 8 weeks, weighing 250-300 g, were used. Six rats were selected and randomly divided into 2 groups ( n=3 each): control group and RPCT group. Rats in control group underwent thoracotomy only, while rats in RPCT group were subjected to an additional 4 cm transverse skin incision along the abdominal midline after thoracotomy. Blood samples were collected, and serum exosomes were isolated from blood samples and labeled as control exosomes and RPCT exosomes. The expression of exosomal surface marker proteins CD9 and heat shock protein 70 (HSP70) was determined by Western blot, and the serum exosome concentration was measured. Another 30 rats were selected and randomly assigned to 5 groups ( n=6 each): sham operation group (Sham group), I/R group, I/R+ RPCT group, I/R+ control exosomes group (I/R+ EXO-CON group), and I/R+ RPCT exosomes group (I/R+ EXO-RPCT group). The MIRI model was established by ligating the left anterior descending coronary artery for 30 min followed by 120 min of reperfusion in anesthetized animals. In I/R+ RPCT group, the MIRI model was prepared at 15 min after the end of RPCT. In I/R+ EXO-CON and I/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 100 μg were administered via the jugular vein at 15 min before ischemia respectively. At the end of reperfusion, the myocardial infarct size was measured, serum concentrations of cardiac troponin T (cTnT) and lactate dehydrogenase (LDH) were determined, and the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in myocardial tissues were measured. The expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3 and cleaved caspase-3 was detected. In vitro experiment H9c2 cells were cultured in vitro and randomly divided into 4 groups ( n=6 each): control group (Con group), hypoxia/reoxygenation (H/R) group, H/R+ control exosomes group (H/R+ EXO-CON group), and H/R+ RPCT exosomes group (H/R+ EXO-RPCT group). The rats were subjected to 4 h of hypoxia followed by 16 h of reoxygenation to establish the H/R injury model. In H/R+ EXO-CON and H/R+ EXO-RPCT groups, control exosomes and RPCT exosomes 4 μg were added at 15 min before hypoxia respectively. The cell survival rate and concentration of lactate dehydrogenase (LDH) in the supernatant were measured, and the expression of Bax, Bcl-2, cleaved caspase-3 and caspase-3 was detected. Results:In vivo experiment Compared with control group, the expression of serum CD9 and HSP70 was significantly up-regulated, and the exosome concentration was increased in RPCT group ( P<0.05). Compared with Sham group, the serum concentrations of cTnT and LDH, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio, and cleaved caspase-3/caspase-3 ratio were significantly increased, and the activity of SOD was decreased in I/R group ( P<0.05). Compared with I/R group, the serum cTnT and LDH concentrations, percentage of myocardial infarct size, content of MDA in myocardial tissues, Bax/Bcl-2 ratio and cleaved caspase-3/caspase-3 ratio were significantly decreased, and the activity of SOD was increased in I/R+ RPCT group ( P<0.05), and no significant changes were observed in the aforementioned parameters in I/R+ EXO-CON group ( P>0.05). There were no significant differences in the aforementioned parameters between I/R+ RPCT group and I/R+ EXO-RPCT group ( P>0.05). In vitro experiment Compared with Con group, the cell survival rate was significantly decreased, and the LDH concentration in the supernatant and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were increased in H/R group ( P<0.05). Compared with H/R group, the cell survival rate was significantly increased, the LDH concentration in the supernatant, and Bax/Bcl-2 and cleaved caspase-3/caspase-3 ratios were decreased in H/R+ EXO-CON group ( P<0.05), and no significant changes were found in the aforementioned parameters in H/R+ EXO-RPCT group ( P>0.05). Conclusions:The mechanism by which RPCT reduces MIRI may be related to the increased release of serum exosomes in rats.
