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[电针预处理对心肌缺血/再灌注损伤大鼠线粒体活性氧介导的细胞凋亡的影响]

[Effect of electroacupuncture preconditioning on cell apoptosis mediated by mitochondrial reactive oxygen species in myocardial ischemia/reperfusion injury rats].

作者信息

Zhang Xiao-Lei, Huang Wei, Yang Qi-Qi, Xiang Li-Li, Xue Yi-Xuan, Lin Yi-Meng, Jiang Gong-Dao

机构信息

College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine/Hubei Provincial Collaborative Innovation Center of Chinese Medicine, Wuhan 430061, China; Department of Acupuncture-moxibustion, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061.

出版信息

Zhen Ci Yan Jiu. 2020 Dec 25;45(12):961-7. doi: 10.13702/j.1000-0607.200053.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) preconditioning on cell apoptosis and the content of mitochondrial reactive oxygen species (ROS) in rats with myocardial ischemia/reperfusion injury (MIRI), so as to explore their mechanisms underlying improvement of MIRI.

METHODS

Eighty male Wistar rats were randomly divided into control, sham, ischemia reperfusion (IR) and EA groups, with 20 rats in each group. Rats of the control, sham and IR groups were just banded on the boards for 30 min, once daily for 7 days. Before modeling, EA (2 Hz, 1 mA) was applied to "Neiguan" (PC6), "Zusanli" (ST36), "Guanyuan"(CV4) for 20 min, once daily for 7 successive days in the EA group. The MIRI model was established by occlusion of the left anterior descending branch of the left coronary artery for 20 min, followed by reperfusion for 30 min. Ventricular arrhythmia (VA) score was used to evaluate arrhythmia. Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of CK-MB. DHE staining was used to detect the content of reactive oxygen species (ROS). The gene expression levels of cytochrome C (Cyt-C), Caspase-9 and Caspase-3 were detected by real-time fluorescent quantitative PCR.

RESULTS

Compared with the sham group, VA score, serum CK-MB content, ROS content in heart tissue and Cyt-C, Caspase-9 and Caspase-3 gene expression were significantly up-regulated in the MIRI group (all <0.01). Following the intervention, the increased VA score, serum CK-MB content, ROS content in heart tissue, Cyt-C, Caspase-9 and Caspase-3 gene expression were all reversed in the EA group compared with the MIRI group (all <0.01).

CONCLUSION

The protective effect of EA preconditioning on MIRI may be based on the regulation of ROS mediated-apoptosis pathway.

摘要

目的

观察电针预处理对心肌缺血/再灌注损伤(MIRI)大鼠细胞凋亡及线粒体活性氧(ROS)含量的影响,以探讨其改善MIRI的机制。

方法

80只雄性Wistar大鼠随机分为对照组、假手术组、缺血再灌注(IR)组和电针组,每组20只。对照组、假手术组和IR组大鼠仅固定于木板上30分钟,每日1次,共7天。电针组在造模前,于“内关”(PC6)、“足三里”(ST36)、“关元”(CV4)施加电针(2Hz,1mA)20分钟,连续7天,每日1次。采用结扎左冠状动脉前降支20分钟,再灌注30分钟的方法建立MIRI模型。采用室性心律失常(VA)评分评估心律失常情况。采用酶联免疫吸附测定(ELISA)法检测肌酸激酶同工酶(CK-MB)水平。采用二氢乙锭(DHE)染色法检测活性氧(ROS)含量。采用实时荧光定量PCR法检测细胞色素C(Cyt-C)、半胱天冬酶-9(Caspase-9)和半胱天冬酶-3(Caspase-3)的基因表达水平。

结果

与假手术组比较,MIRI组VA评分、血清CK-MB含量、心脏组织ROS含量及Cyt-C、Caspase-9和Caspase-3基因表达均显著上调(均P<0.01)。干预后,与MIRI组比较,电针组升高的VA评分、血清CK-MB含量、心脏组织ROS含量及Cyt-C、Caspase-9和Caspase-3基因表达均降低(均P<0.01)。

结论

电针预处理对MIRI的保护作用可能基于对ROS介导的凋亡通路的调控。

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