Mechanism of the pretreatment with electroacupuncture of "<italic style="font-style: italic">biaoben</italic> acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury

ZHANG Yanlin; WU Song; GUO Qianru; YU Yuntao; WANG Sunyi; WEI Yuqi; WAN Xiaoman; LU Zhen; HE Xiaoru

doi:10.13703/j.0255-2930.20240317-k0004

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Mechanism of the pretreatment with electroacupuncture of "biaoben acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury

Study on Mechanism | 更新时间：2025-03-13

- Mechanism of the pretreatment with electroacupuncture of "biaoben acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury
- Chinese Acupuncture & Moxibustion Vol. 45, Issue 3, Pages: 335-344(2025)
- 作者机构：
  
  湖北中医药大学针灸骨伤学院/针灸治未病湖北省协同创新中心，武汉 430061
  湖北时珍实验室，武汉 430061
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20240317-k0004
  CLC：
- Received：17 March 2024，
  
  Online First：22 August 2024，
  
  Published：12 March 2025
- 稿件说明：
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ZHANG Yanlin, WU Song, GUO Qianru, et al. Mechanism of the pretreatment with electroacupuncture of "biaoben acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury[J]. Chinese Acupuncture & Moxibustion, 2025, 45(3): 335-344. DOI: 10.13703/j.0255-2930.20240317-k0004.
DOI：

ZHANG Yanlin, WU Song, GUO Qianru, et al. Mechanism of the pretreatment with electroacupuncture of "biaoben acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury[J]. Chinese Acupuncture & Moxibustion, 2025, 45(3): 335-344. DOI: 10.13703/j.0255-2930.20240317-k0004. DOI：

摘要

目的：

观察“标本配穴”电针预处理对心肌缺血再灌注损伤（MIRI）大鼠心肌细胞线粒体分裂的影响，并探讨其作用机制。

方法：

将50只雄性SD大鼠随机分为假手术组、模型组、电针预处理组、电针预处理+Compound C组和电针预处理+ML385组，各10只。电针预处理组、电针预处理+Compound C组和电针预处理+ML385组于双侧“内关”“足三里”和“关元”行电针干预20 min，连续波，频率2 Hz，电流1 mA，每日1次，连续7 d。第8天，电针预处理+Compound C组和电针预处理+ML385组于造模前30 min分别腹腔注射Compound C溶液（0.3 mg/kg）和ML385溶液（30 mg/kg）。除假手术组外，其余组大鼠均采用冠状动脉左前降支结扎法建立MIRI大鼠模型，假手术组穿线不结扎。造模结束后，采用流式细胞仪检测缺血区心肌组织活性氧（ROS）含量，黄嘌呤氧化酶法检测缺血区心肌组织超氧化物歧化酶（SOD）含量，硫代巴比妥酸比色法检测缺血区心肌组织丙二醛（MDA）含量，HE染色法观察缺血区心肌组织形态，透射电镜观察缺血区心肌细胞线粒体超微结构，免疫荧光法检测缺血区心肌组织线粒体分裂因子（MFF）、线粒体裂变1蛋白抗体（Fis1）和动力蛋白相关蛋白1（Drp1）阳性表达，免疫组织化学法检测缺血区心肌组织腺苷酸活化蛋白激酶（AMPK）、核因子E2相关因子2（Nrf2）和Drp1蛋白表达。

结果：

与假手术组比较，模型组缺血区心肌组织ROS、MDA含量及MFF、Fis1、Drp1阳性表达均升高（

0.01），SOD含量及AMPK、Nrf2蛋白表达均降低（

0.01），Drp1蛋白表达升高（

0.01）。与模型组比较，电针预处理组缺血区心肌组织ROS、MDA含量及MFF、Fis1、Drp1阳性表达均降低（

0.01），SOD含量及AMPK、Nrf2蛋白表达均升高（

0.01），Drp1蛋白表达降低（

0.01）；电针预处理+Compound C组和电针预处理+ML385组缺血区心肌组织MFF、Fis1、Drp1阳性表达及Drp1蛋白表达均降低（

0.01）。与电针预处理+Compound C组和电针预处理+ML385组比较，电针预处理组缺血区心肌组织ROS、MDA含量及MFF、Fis1、Drp1阳性表达均降低（

0.01），SOD含量及AMPK、Nrf2蛋白表达升高（

0.01

0.05），Drp1蛋白表达降低（

0.05）。与模型组、电针预处理+Compound C组和电针预处理+ML385组比较，电针预处理组心肌纤维断裂程度、细胞肿胀程度、线粒体紊乱破碎程度均明显减轻，模型组、电针预处理+Compound C组和电针预处理+ ML385组心肌纤维断裂程度、细胞肿胀程度、线粒体紊乱破碎程度较为接近。

结论：

“标本配穴”电针预处理可能通过促进AMPK和Nrf2磷酸化，抑制Drp1诱导的线粒体过度分裂，减轻线粒体碎片化和空泡化引起的线粒体功能障碍，从而减轻MIRI大鼠心肌损伤。

Abstract

Objective

To observe the effect of electroacupuncture (EA) pretreatment of "

biaoben

acupoint combination" on cardiomyocyte mitochondrial fission in the rats with myocardial ischemia-reperfusion injury (MIRI) and explore its mechanism.

Methods

Fifty male SD rats were randomly divided into a sham-operation group

a model group

an EA pretreatment group

an EA pretreatment + Compound C group and an EA pretreatment+ML385 group

10 rats in each group. In the EA pretreatment

the EA pretreatment + Compound C group and the EA pretreatment+ML385 group

EA was delivered at bilateral "Neiguan" (PC6)

"Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min

with continuous wave and 2 Hz of frequency

1 mA of current

once daily for consecutive 7 days. On day 8

in the EA pretreatment + Compound C group and the EA pretreatment+ML385 group

30 min before model preparation

the intraperitoneal injection with Compound C (0.3 mg/kg) and ML385 (30 mg/kg) was administered respectively. Except in the sham-operation group

the ligation of the left anterior descending coronary artery was performed to prepare MIRI rat model in the rest groups. In the sham-operation group

the thread was not ligated. After modeling

the content of reactive oxygen species (ROS) in the ischemic area was measured by flow cytometry

superoxide dismutase (SOD) was detected using xanthine oxidase method

and malondialdelyde (MDA) was detected using thiobarbituric acid (TBA) chromatometry. The morphology of myocardial tissue in the ischemic area was observed with HE staining

and the mitochondria ultrastructure of cardiomyocytes observed under transmission electron microscopy. Using immunofluorescence analysis

the positive expression of mitochondrial fission factor (MFF)

mitochondrial fission 1 protein antibody (Fis1) and dynamin-related protein 1 (Drp1) was detected; and with immunohistochemical method used

the protein expression of adenosine monophosphate-activated protein kinase (AMPK)

nuclear factor E2-associated factor2 (Nrf2) and Drp1 in the ischemic area was detected.

Results

Compared with the sham-operation group

the content of ROS and MDA in the myocardial tissue of the ischemic area

and the positive expression of MFF

Fis1 and Drp1 increased in the model group (

0.01); the content of SOD and the protein expression of AMRK and Nrf2 decreased (

0.01)

and the protein expression of Drp1 elevated (

0.01). Compared with the model group

the content of ROS and MDA in the myocardial tissue of the ischemic area

and the positive expression of MFF

Fis1 and Drp1 were dropped in the EA pretreatment group (

0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (

0.01)

and the protein expression of Drp1 declined (

0.01); and in the EA pretreatment+Compound C group and the EA pretreatment+ML385 group

the positive expression of MFF

Fis1 and Drp1

and the protein expression of Drp1 were all reduced (

0.01). When compared with the EA pretreatment + Compound C group and the EA pretreatment+ML385 group

the content of ROS and MDA in the myocardial tissue of the ischemic area

and the positive expression of MFF

Fis1 and Drp1 were dropped in the EA pretreatment group (

0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (

0.01

0.05)

and the protein expression of Drp1 decreased (

0.05). In comparison with the model group

the EA pretreatment+Compound C

group and the EA pretreatment+ML385 group

the cardiac muscle fiber rupture

cell swelling and mitochondrial disorders were obviously alleviated in the EA pretreatment group. The morphological changes were similar among the model group

the EA pretreatment+Compound C group and the EA pretreatment+ML385 group.

Conclusion

Electroacupuncture pretreatment of "

biaoben

acupoint combination" attenuates myocardial injury in MIRI rats

probably through promoting the phosphorylation of AMPK and Nrf2

inhibiting the excessive mitochondrial fission induced by Drp1

and reducing mitochondrial dysfunction caused by mitochondrial fragmentation and vacuolation.

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Related Author

WEI Yuqi

ZHANG Yanlin

WAN Xiaoman

MAO Jing

WANG Sunyi

JIANG Chunyan

CAO Zhenyang

GUO Qianru

Related Institution

College of Acupuncture-Moxibustion and Orthopedics, Hubei University of CM, Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture and Moxibustion

Department of Acupuncture-Moxibustion, Affiliated Hospital of Hubei University of CM, Hubei Provincial Hospital of TCM

School of Acupuncture-Moxibustion and Tuina, Beijing University of CM

Department of Orthopedics, Guangyuan Second People's Hospital

Department of General Medicine, Guangyuan Maternal and Child Health Hospital

⁰

Mechanism of the pretreatment with electroacupuncture of "biaoben acupoint combination" for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury

DOI：10.13703/j.0255-2930.20240317-k0004

摘要

Abstract

关键词

Keywords

references