心肌缺血预适应循环血中微囊泡对大鼠心肌I/R损伤的作用

doi:10.13459/j.cnki.cjap.2016.02.001

中国应用生理学杂志 ›› 2016, Vol. 32 ›› Issue (2): 97-101.doi: 10.13459/j.cnki.cjap.2016.02.001

• 研究论文 • 下一篇

心肌缺血预适应循环血中微囊泡对大鼠心肌I/R损伤的作用

王艺璐, 刘淼, 尚曼, 王瑶, 张琦, 王少勋, 韦苏, 张琨玮, 刘超, 吴艳娜, 宋君秋, 刘艳霞

天津医科大学基础医学院药理学教研室, 天津 300070

收稿日期:2015-12-14 修回日期:2016-02-16 出版日期:2016-03-28 发布日期:2018-06-12
通讯作者: 宋君秋,Tel/Fax:86-22-83336660;E-mail:songjunqiu@126.com;刘艳霞,Tel/Fax:86-22-83336660;E-mail:liu_yanxia126@126.com E-mail:songjunqiu@126.com;liu_yanxia126@126.com
基金资助:
天津市自然科学基金项目（11JCZDJC18300）；高校博士学科点专项科研基金项目（20101202110005）；天津市高等学校科技发展基金计划项目（20110106）

Effects of circulating microvesicles derived from myocardial ischemic preconditioning on myocardial ischemia/reperfusion injury in rats

WANG Yi-lu, LIU Miao, SHANG Man, WANG Yao, ZHANG Qi, WANG Shao-xun, WEI Su, ZHANG Kun-wei, LIU Chao, WU Yan-na, SONG Jun-qiu, LIU Yan-xia

Department of Pharmacology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China

Received:2015-12-14 Revised:2016-02-16 Online:2016-03-28 Published:2018-06-12
Supported by:
天津市自然科学基金项目（11JCZDJC18300）；高校博士学科点专项科研基金项目（20101202110005）；天津市高等学校科技发展基金计划项目（20110106）

摘要/Abstract

摘要： 目的：研究心肌缺血预适应（IPC）大鼠循环血中微囊泡（MVs）对大鼠在体心肌缺血/再灌注（I/R）损伤的作用及相关机制。方法：反复短暂结扎/松开大鼠冠状动脉左前降支建立大鼠IPC模型，自腹主动脉取血，超速离心法分离循环血中的IPC-MVs，并对其进行流式鉴定。建立在体大鼠心肌I/R模型，股静脉注射IPC-MVs 7 mg/kg。HE染色观察心肌形态学变化，TTC染色检测心肌梗死范围，TUNEL染色检测心肌细胞凋亡率。比色法测定血清乳酸脱氢酶（LDH）活力，分光光度法测定心肌组织caspase 3活力，Western blot法检测心肌组织Bcl-2、Bax蛋白表达水平。结果：流式细胞术检测IPC-MVs浓度为4380±745个/μl。与I/R组比较，IPC-MVs能够减轻I/R大鼠心肌组织损伤，缩小心肌梗死范围（P<0.01），减少心肌细胞凋亡数量（P<0.01），明显降低血清LDH活力（P<0.01），降低心肌组织caspase 3活力（P<0.01），升高Bcl-2蛋白表达（P<0.01），降低Bax蛋白表达（P<0.01），升高Bcl-2/Bax比值（P<0.01）。结论：IPC-MVs显著减轻大鼠在体心肌I/R损伤，通过上调心肌组织中Bcl-2的蛋白表达，下调Bax的蛋白表达，升高Bcl-2/Bax比值，降低caspase 3活力而发挥心肌保护作用。

关键词: 细胞微囊泡, 心肌缺血预适应, 心肌缺血/再灌注, 大鼠, 细胞凋亡

Abstract: Objective: To investigate the effects of circulating microvesicles (MVs) derived from ischemic preconditioning (IPC) on myocardial ischemia/reperfusion (I/R) injury in rats and explore the underlying mechanism. Methods: To establish the IPC model, the rats were subjected to brief cycles of left anterior descending (LAD) coronary occlusion and reperfusion. The blood was drawn from abdominal aorta once the operation was finished. IPC-MVs were isolated by ultracentrifugation from the peripheral blood and characterized by flow cytometry. The myocardial I/R model of rats was established in vivo. Rats were injected via the femoral vein with IPC-MVs at 7 mg/kg. Morphological changes of myocardium were observed microscopically after HE staining. Apoptosis of myocardial cells was detected with TUNEL assay. Myocardial infarct size was detected by TTC staining. Moreover, activity of plasma lactate dehydrogenase (LDH) was tested by colorimetry. The activity of caspase 3 in myocardium was assayed with spectrophotometry. Expression levels of Bcl-2 and Bax protein were examined with Western blot. Results: The concentration of IPC-MVs, which was detected by flow cytometry, was 4380±745 cells/μl. Compared with I/R group, IPC-MVs alleviated the damage of tissues in I/R injured rats significantly. The myocardial infarct size and the cardiomyocyte apoptotic index were obviously decreased after IPC-MVs treatment (P<0.01, respectively). The activity of plasma LDH was significantly decreased in IPC-MVs treated rats (P<0.01). Moreover, the activity of caspase 3 was markedly decreased after IPC-MVs treatment (P<0.01). In addition, the expression of Bcl-2 was increased (P<0.01), the expression of Bax was decreased (P<0.01), the ratio of Bcl-2/Bax was significantly increased after IPC-MVs treatment (P<0.01). Conclusion: IPC-MVs protected myocardial against I/R injury by up-regulating the expression of Bcl-2 protein, down-regulating the expression of Bax protein, increasing the ratio of Bcl-2/Bax and decreasing cleavage of caspase 3.

Key words: microvesicles, myocardial ischemic preconditioning, myocardial ischemia/reperfusion, rats, apoptosis

王艺璐, 刘淼, 尚曼, 王瑶, 张琦, 王少勋, 韦苏, 张琨玮, 刘超, 吴艳娜, 宋君秋, 刘艳霞. 心肌缺血预适应循环血中微囊泡对大鼠心肌I/R损伤的作用[J]. 中国应用生理学杂志, 2016, 32(2): 97-101.

WANG Yi-lu, LIU Miao, SHANG Man, WANG Yao, ZHANG Qi, WANG Shao-xun, WEI Su, ZHANG Kun-wei, LIU Chao, WU Yan-na, SONG Jun-qiu, LIU Yan-xia. Effects of circulating microvesicles derived from myocardial ischemic preconditioning on myocardial ischemia/reperfusion injury in rats[J]. CJAP, 2016, 32(2): 97-101.

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心肌缺血预适应循环血中微囊泡对大鼠心肌I/R损伤的作用

Effects of circulating microvesicles derived from myocardial ischemic preconditioning on myocardial ischemia/reperfusion injury in rats

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