基于核受体辅激活因子 4 介导的铁蛋白自噬探讨针刺减轻脑缺血再灌注损伤的机制。

Mechanism of acupuncture in attenuating cerebral ischaemia-reperfusion injury based on nuclear receptor coactivator 4 mediated ferritinophagy.

机构信息

College of Acupuncture-Moxibustion and Tuina, Nanjing University of Chinese Medicine, Nanjing 210023, China.

2 Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing 210023, China.

出版信息

J Tradit Chin Med. 2024 Apr;44(2):345-352. doi: 10.19852/j.cnki.jtcm.20240203.006.

DOI:10.19852/j.cnki.jtcm.20240203.006

PMID:38504540

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10927404/

Abstract

OBJECTIVE

To explore the effect of acupuncture treatment on cerebral ischaemia-reperfusion injury (CIRI) and reveal the underlying mechanism of the effect based on nuclear receptor coactivator 4 (NCOA4) mediated ferritinophagy.

METHODS

Sprague-Dawley male rats were divided into four groups: the sham group, model group, acupuncture group, and sham acupuncture group. After 2 h of middle cerebral artery occlusion (MCAO), reperfusion was performed for 24 h to induce CIRI. The rats were treated with acupuncture at the Neiguan (PC6) and Shuigou (GV26) acupoints. Their neurological function was evaluated by taking their Bederson scores at 2 h after ischaemia and 24 h after reperfusion. Triphenyltetrazolium chloride staining was applied to assess the cerebral infarct volume at 24 h after reperfusion. The malondialdehyde (MDA) and ferrous iron (Fe) levels were observed after 24 h of reperfusion using an assay kit. Western blotting was performed to detect the expression of NCOA4 and ferritin heavy chain 1 (FTH1) at 24 h after reperfusion. Moreover, the colocalization of ferritin with neurons, NCOA4 with microtubule-associated protein 1 light chain 3 (LC3), and NCOA4 with ferritin was visualized using immunofluorescence staining.

RESULTS

Acupuncture significantly improved neurological function and decreased cerebral infarct volume in the acupuncture group. Following CIRI, the expression of NCOA4, LC3 and FTH1 was increased, which enhanced ferritinophagy and induced an inappropriate accumulation of Fe and MDA in the ischaemic brain. However, acupuncture dramatically downregulated the expression of NCOA4, LC3 and FTH1, inhibited the overactivation of ferritinophagy, and decreased the levels of MDA and Fe.

CONCLUSIONS

Acupuncture can inhibit NCOA4-mediated ferritinophagy and protect neurons against CIRI in a rat model.

摘要

目的

探讨针刺治疗对脑缺血再灌注损伤（CIRI）的影响，并基于核受体共激活因子 4（NCOA4）介导的铁蛋白自噬来揭示其作用机制。

方法

将雄性 Sprague-Dawley 大鼠分为四组：假手术组、模型组、针刺组和假针刺组。在大脑中动脉闭塞（MCAO）后 2 h 进行再灌注，以诱导 CIRI。再灌注 24 h 后，采用针刺内关（PC6）和水沟（GV26）穴对大鼠进行治疗。在缺血后 2 h 和再灌注后 24 h 时，通过Bederson 评分评估大鼠的神经功能。再灌注 24 h 后，采用氯化三苯基四氮唑染色评估脑梗死体积。通过试剂盒检测再灌注 24 h 后丙二醛（MDA）和亚铁（Fe）的水平。再灌注 24 h 后，通过 Western blot 检测 NCOA4 和铁蛋白重链 1（FTH1）的表达。此外，采用免疫荧光染色观察铁蛋白与神经元、NCOA4 与微管相关蛋白 1 轻链 3（LC3）、NCOA4 与铁蛋白的共定位。