井穴放血通过激活HIF-1α/BNIP3信号通路介导的线粒体自噬和降低氧化应激减轻急性高原低氧大鼠心肌损伤

WANG Chao; LI Meng-xin; LI Yun-di; LI Yong-ping

doi:10.1007/s11655-022-3626-4

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井穴放血通过激活HIF-1α/BNIP3信号通路介导的线粒体自噬和降低氧化应激减轻急性高原低氧大鼠心肌损伤

Acupuncture Research | Updated：2023-02-01

- 井穴放血通过激活HIF-1α/BNIP3信号通路介导的线粒体自噬和降低氧化应激减轻急性高原低氧大鼠心肌损伤
- Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress
- 中国结合医学杂志（英文版） 2023年29卷第2期页码：170-178
- Affiliations：
  
  Department of Traditional Chinese Medicine, Medical Institute of Qinghai University, Xining (810000), China
- Author bio：
  
  Prof. LI Yong-ping, E-mail: 2007980002@qhu.edu.cn
- Funds：
  
  the Applied Basic Research Project of Science and Technology Department of Qinghai Province(2020-ZJ-760)
- DOI：10.1007/s11655-022-3626-4
  中图分类号：
- 纸质出版日期：2023-02，
  
  网络出版日期：2022-12-09，
  
  录用日期：2022-08-26
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井穴放血通过激活HIF-1α/BNIP3信号通路介导的线粒体自噬和降低氧化应激减轻急性高原低氧大鼠心肌损伤[J]. 中国结合医学杂志（英文版）, 2023,29(2):170-178.

WANG Chao, LI Meng-xin, LI Yun-di, et al. Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress[J]. Chinese Journal of Integrative Medicine, 2023,29(2):170-178.
井穴放血通过激活HIF-1α/BNIP3信号通路介导的线粒体自噬和降低氧化应激减轻急性高原低氧大鼠心肌损伤[J]. 中国结合医学杂志（英文版）, 2023,29(2):170-178. DOI： 10.1007/s11655-022-3626-4.

WANG Chao, LI Meng-xin, LI Yun-di, et al. Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress[J]. Chinese Journal of Integrative Medicine, 2023,29(2):170-178. DOI： 10.1007/s11655-022-3626-4.

摘要

目的:

探讨井穴放血对急性低压缺氧 (AHH) 大鼠心肌损伤的保护作用及其可能机制.

方法:

75只大鼠按随机数字表随机分为5组: 对照组 (

=15) 、模型组 (

=15) 、BAJP组 (

=15) 、BAJP+3-甲基腺嘌呤 (3-MA) 组 (

=15) 和BANA组 (非穴位针刺放血

尾部放血

n=15) . 除对照组外

其余各组均建立AHH大鼠模型

并采用相应的治疗方法. 采用酶联免疫吸附试验 (ELISA) 检测血清肌酸激酶同工酶MB (CK-MB) 和心肌肌钙蛋白I (CTnI) 水平以及心肌组织超氧化物歧化酶 (SOD) 和丙二醛 (MDA) 水平. 苏木精-伊红 (HE) 染色观察心肌损伤

末端脱氧核苷酸转移酶介导的dUTP生物素缺口末端标记 (TUNEL) 染色观察细胞凋亡. 透射电镜观察心肌线粒体损伤和自噬体. JC-1荧光染色法分析心肌线粒体膜电位. 线粒体呼吸链复合物测定试剂盒检测心肌线粒体呼吸链复合体 (Ⅰ、Ⅲ和Ⅳ) 活性和ATP酶. Western blot分析检测自噬指标、缺氧诱导因子-1α (HIF-1α) /Bcl-2和腺病毒E1B 19k Da相互作用蛋白3 (BNIP3) 信号通路.

结果:

BAJP减轻AHH大鼠心肌损伤

抑制心肌细胞凋亡. BAJP预处理降低了AHH大鼠的MDA水平

增加了SOD水平 (均

0.01) . 此外

BAJP预处理增加了AHH大鼠的线粒体膜电位 (

0.01) 、线粒体呼吸链复合体 (复合体Ⅰ、Ⅲ和Ⅳ) 活性 (

0.01) 和线粒体ATP酶活性 (

0.05) . 电子显微镜的结果表明

BAJP预处理改善了AHH大鼠心肌线粒体肿胀并增加了自噬体数量. 此外

BAJP预处理激活了HIF-1α/BNIP3通路和自噬. 最后

在BAJP治疗的AHH大鼠中使用3-MA抑制自噬的结果表明

自噬的抑制减弱了BAJP对AHH大鼠的治疗效果

进一步证明自噬是BAJP治疗AHH的潜在靶点.

结论:

BAJP是AHH诱导心肌损伤的有效治疗方法

其机制可能与增加HIF-1α/BNIP3信号介导的自噬和降低氧化应激有关.

Abstract

Objective:

To explore the protective effect and possible mechanisms of bloodletting acupuncture at Jing-well points (BAJP) pre-treatment on acute hypobaric hypoxia (AHH)-induced myocardium injury rat.

Methods:

Seventy-five rats were randomly divided into 5 groups by a random number table: a control group (

=15)

a model group (

=15)

a BAJP group (

=15)

a BAJP+3-methyladenine (3-MA) group (

=15)

and a BANA (bloodletting at nonacupoint; tail bleeding

=15) group. Except for the control group

the AHH rat model was established in the other groups

and the corresponding treatment methods were adopted. Enzyme-linked immunosorbent assay (ELISA) was used to detect creatine kinase isoenzyme MB (CK-MB) and cardiac troponins I (CTnI) levels in serum and superoxide dismutase (SOD) and malondialdehyde (MDA) levels in myocardial tissue. Hematoxylin-eosin (HE) staining was used to observe myocardial injury

and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to observe cell apoptosis. Transmission electron microscopy detection was used to observe mitochondrial damage and autophagosomes in the myocardium. The mitochondrial membrane potential of the myocardium was analyzed with the fluorescent dye JC-1. Mitochondrial respiratory chain complex (complex Ⅰ

Ⅲ

and Ⅳ) activities and ATPase in the myocardium were detected by mitochondrial respiratory chain complex assay kits. Western blot analysis was used to detect the autophagy index and hypoxia inducible factor-1α (HIF-1α)/Bcl-2 and adenovirus E1B 19k Da-interacting protein 3 (BNIP3) signaling.

Results:

BAJP reduced myocardial injury and inhibited myocardial cell apoptosis in AHH rats. BAJP pretreatment decreased MDA levels and increased SOD levels in AHH rats (all

0.01). Moreover

BAJP pretreatment increased the mitochondrial membrane potential (

0.01)

mitochondrial respiratory chain complex (complexes Ⅰ

Ⅲ

and Ⅳ) activities (

0.01)

and mitochondrial ATPase activity in AHH rats (

0.05). The results from electron microscopy demonstrated that BAJP pretreatment improved mitochondrial swelling and increased the autophagosome number in the myocardium of AHH rats. In addition

BAJP pretreatment activated the HIF-1α/BNIP3 pathway and autophagy. Finally

the results of using 3-MA to inhibit autophagy in BAJP-treated AHH rats showed that suppression of autophagy attenuated the treatment effects of BAJP in AHH rats

further proving that autophagy constitutes a potential target for BAJP treatment of AHH.

Conclusion:

BAJP is an effective treatment for AHH-induced myocardial injury

and the mechanism might involve increasing HIF-1α/BNIP3 signaling-mediated autophagy and decreasing oxidative stress.

关键词

急性低压低氧针刺井穴放血自噬心肌损伤线粒体损伤中医

Keywords

acute hypobaric hypoxiabloodletting acupuncture at Jing-well pointsautophagymyocardium injurymitochondrial damageChinese medicine

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