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[电针预处理对大鼠心肌缺血再灌注损伤的保护作用及对FXR/SHP基因的调控作用]

[Protective effect and regulating effect on FXR/SHP gene of electroacupuncture preconditioning on myocardial ischemia-reperfusion injury in rats].

作者信息

Li Chen, Zhang Xiao-Lei, Xue Yi-Xuan, Cheng De-Jun, Yan Jiang-Tian, Wu Song, Huang Wei

机构信息

College of Acupuncture-Moxibustion and Orthopaedics, Hubei University of CM/ Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Disease with Acupuncture and Moxibustion, Wuhan 430061, China; Third People's Hospital of Hubei Province, Wuhan 430033.

College of Acupuncture-Moxibustion and Orthopaedics, Hubei University of CM/ Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Disease with Acupuncture and Moxibustion, Wuhan 430061, China; Department of Acupuncture and Moxibustion, Hubei Provincial Hospital of TCM, Wuhan 430061.

出版信息

Zhongguo Zhen Jiu. 2019 Aug 12;39(8):861-6. doi: 10.13703/j.0255-2930.2019.08.016.

Abstract

OBJECTIVE

To explore the protective mechanism of electroacupuncture preconditioning on myocardial ischemia-reperfusion injury in rats.

METHODS

A total of 88 SPF-grade Wistar male rats were randomized into a normal group, a sham-operation group, a model group and an electroacupuncture (EA) group, 22 rats in each one. The rats in the normal group received normal diet and no intervention was given for 7 d. The rats in the sham-operation and the model group were bound for 20 min, once a day for 7 d. In the EA group, EA, continuous wave, 2 Hz and 1 mA, was applied at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) for 20 min, once a day for 7 d. In the 8th day, the blood was collected by abdominal aortic method 60 min after intraperitoneal injection of 10% urethane anesthesia by 10 mL/kg in the normal group, and then the tissue of heart was collected. In the model group and the EA group, ligating the left anterior descending coronary artery (LAD) was adopted for 20 min and reperfusion was applied for 40 min, then the samples were collected. The rats in the sham-operation group recevied open-chest operation and no other intervention, the samples were collected in 60 min. Nitro Blue Tetrazolium Chloride monohydrate (NBT) stain was used to detected myocardial infarct size and weight, ELISA was used to measured myocardial injury markers and inflammatory factors (LDH、CK、cTnI), and the fluorescent quantitative PCR method was used to determine the expressions of FXR and SHP gene.

RESULTS

①There was no significant difference between the normal group and the sham-operation group in myocardial infarct size and weight (>0.05) .Compared with the normal group, myocardial infarct size and weight were higher in the model group (<0.01). Compared with the model group, myocardial infarct size and weight were lower in the EA group (<0.01). ②There was no significant difference between the normal group and the sham-operation group in serum contents of LDH、CK、cTnI (>0.05). Compared with the normal group, the serum contents of LDH、CK、cTnI were higher in the model group (<0.05). Compared with the model group, the serum contents of LDH、CK、cTnI were lower in the EA group (<0.05). ③The expressions of FXR and SHP gene in the model group were higher than those in the normal group (<0.05). The expressions of FXR and SHP gene in the EA group were lower than those in the model group (<0.05).

CONCLUSION

Electroacupuncture preconditioning could significantly improve cardiac function of rats with myocardial ischemia-reperfusion, reduce infarct size and inflammatory factors, and down-regulate the expressions of FXR and SHP gene. The protective effect may generate based on regulation of FXR/SHP apoptosis signal pathway.

摘要

目的

探讨电针预处理对大鼠心肌缺血再灌注损伤的保护机制。

方法

将88只SPF级雄性Wistar大鼠随机分为正常组、假手术组、模型组和电针组,每组22只。正常组大鼠给予正常饮食,7天不做干预。假手术组和模型组大鼠每天束缚20分钟,连续7天。电针组大鼠在“内关”(PC 6)、“足三里”(ST 36)和“关元”(CV 4)穴位施加连续波、2Hz、1mA的电针刺激20分钟,每天1次,连续7天。第8天,正常组大鼠腹腔注射10%水合氯醛10 mL/kg麻醉后60分钟,经腹主动脉采血,然后取心脏组织。模型组和电针组大鼠采用结扎左冠状动脉前降支(LAD)20分钟,再灌注40分钟,然后采集样本。假手术组大鼠进行开胸手术,不做其他干预,60分钟后采集样本。采用氯化硝基四氮唑蓝(NBT)染色检测心肌梗死面积和重量,采用酶联免疫吸附测定法(ELISA)检测心肌损伤标志物和炎症因子(LDH、CK、cTnI),采用荧光定量PCR法检测FXR和SHP基因的表达。

结果

①正常组与假手术组心肌梗死面积和重量比较,差异无统计学意义(>0.05)。与正常组比较,模型组心肌梗死面积和重量升高(<0.01)。与模型组比较,电针组心肌梗死面积和重量降低(<0.01)。②正常组与假手术组血清LDH、CK、cTnI含量比较,差异无统计学意义(>0.05)。与正常组比较,模型组血清LDH、CK、cTnI含量升高(<0.05)。与模型组比较,电针组血清LDH、CK、cTnI含量降低(<0.05)。③模型组FXR和SHP基因表达高于正常组(<0.05)。电针组FXR和SHP基因表达低于模型组(<0.05)。

结论

电针预处理可显著改善心肌缺血再灌注大鼠的心功能,减小梗死面积,降低炎症因子水平,下调FXR和SHP基因表达。其保护作用可能是通过调节FXR/SHP凋亡信号通路产生的。

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