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[再灌注不同时间电针干预对心肌缺血再灌注损伤大鼠心肌组织Bcl-2及Beclin1表达的影响]

[Effects of electroacupuncture at different time during reperfusion on the expression of Bcl-2 and Beclin1 in myocardial tissue in rats with myocardial ischemia reperfusion injury].

作者信息

Zhang Hongru, Zhong Zehao, Chen Wanying, Bai Hua, Xiao Yan, Gu Yihuang, Lu Shengfeng

机构信息

The Second Clinical Medical College of Nanjing University of CM, Nanjing 210023, Jiangsu Province, China.

The Second Clinical Medical College of Nanjing University of CM, Nanjing 210023, Jiangsu Province, China; Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of CM, Nanjing 210023, Jiangsu Province.

出版信息

Zhongguo Zhen Jiu. 2018 Nov 12;38(11):1195-200. doi: 10.13703/j.0255-2930.2018.11.015.

Abstract

OBJECTIVE

To compare the effects of electroacupuncture (EA) at different time during reperfusion on the expression of autophagy-related protein Bcl-2 and Beclin1 in myocardial tissue in rats with myocardial ischemia reperfusion injury (MIRI), and to explore the autophagy-related mechanism of EA on protecting MIRI.

METHODS

A total of 72 SD rats were randomly divided into a sham operation group, a model group, a RA group, a RB group, a RC group and a RD group, 12 rats in each group. Except the sham operation group, the rats in the remaining groups were treated with ligating the left anterior descending artery (LAD) for 30 minutes followed by reperfusion to establish the model of MIRI. The rats in the sham operation group were treated with crossing a line through the LAD. The rats in the model group did not receive treatment. The rats in the RA group, RB group, RC group and RD group were treated with EA at "Neiguan" (PC 6) for 20 min, starting at 0 h, 0.5 h, 1 h, 2 h after reperfusion. Evans Blue-TTC double-staining was employed to evaluate myocardial infarct size; the serum CK-MB was detected by ELISA and the expression of Bcl-2 and Beclin1 protein in myocardial tissue were detected by Western blot.

RESULTS

Compared with the model group, the percentage of myocardial infarct size in the RB group, RC group and RD group was decreased significantly (all <0.05), and the reduction in the RB group was more significant than that in the RC group and RD group (both <0.05). Compared with the sham operation group, the expression of CK-MB and Beclin1 in the model group was significantly increased, but the expression of Bcl-2 was significantly decreased (all <0.01). Compared with the model group, the expression of CK-MB and Beclin1 was decreased significantly in RA group, RB group, RC group and RD group (all <0.05), but the expression of Bcl-2 was significantly increased (all <0.05). Compared with the RA group, the expression of CK-MB was decreased in the RB group and RC group (both <0.05) but the expression of Bcl-2 was increased (both <0.01); the expression was not significantly different from that in the RD group (>0.05); the increasing of Bcl-2 in the RB group was more significant than that in RC group (<0.05). The expression of Beclin 1 in the RB group was significantly lower than that in the RA group (<0.05), but there was no significant difference among other EA groups (all >0.05).

CONCLUSION

EA at different time during reperfusion could reduce myocardial infarct size in rats with MIRI, and EA at 0.5 h after reperfusion has best efficacy; this protective effect may be achieved by increasing Bcl-2 expression and reducing Beclin1 expression to inhibit overautophagy during reperfusion.

摘要

目的

比较电针(EA)在再灌注不同时间对心肌缺血再灌注损伤(MIRI)大鼠心肌组织自噬相关蛋白Bcl-2和Beclin1表达的影响,探讨EA保护MIRI的自噬相关机制。

方法

将72只SD大鼠随机分为假手术组、模型组、RA组、RB组、RC组和RD组,每组12只。除假手术组外,其余各组大鼠采用结扎左冠状动脉前降支(LAD)30分钟后再灌注的方法建立MIRI模型。假手术组大鼠仅穿线通过LAD。模型组大鼠不接受治疗。RA组、RB组、RC组和RD组大鼠于再灌注后0小时、0.5小时、1小时、2小时开始在“内关”(PC 6)穴进行20分钟电针治疗。采用伊文思蓝-氯化三苯基四氮唑(Evans Blue-TTC)双重染色法评估心肌梗死面积;采用酶联免疫吸附测定法(ELISA)检测血清肌酸激酶同工酶(CK-MB)水平,采用蛋白质免疫印迹法检测心肌组织中Bcl-2和Beclin1蛋白的表达。

结果

与模型组比较,RB组、RC组和RD组心肌梗死面积百分比均显著降低(均<0.05),且RB组降低幅度大于RC组和RD组(均<0.05)。与假手术组比较,模型组CK-MB和Beclin1表达显著升高,而Bcl-2表达显著降低(均<0.01)。与模型组比较,RA组、RB组、RC组和RD组CK-MB和Beclin1表达均显著降低(均<0.05),而Bcl-2表达均显著升高(均<0.05)。与RA组比较,RB组和RC组CK-MB表达降低(均<0.05),Bcl-2表达升高(均<0.01);与RD组比较,差异无统计学意义(>0.05);RB组Bcl-2升高幅度大于RC组(<0.05)。RB组Beclin 1表达显著低于RA组(<0.05),其他电针组间差异无统计学意义(均>0.05)。

结论

再灌注不同时间电针均可减小MIRI大鼠心肌梗死面积,再灌注后0.5小时电针疗效最佳;其保护作用可能是通过上调Bcl-2表达、下调Beclin1表达,抑制再灌注期过度自噬实现的。

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