Mechanism of Mitochondrial Autophagy in Cerebral Tissue of Ischemic Stroke Rats Through HIF-1α/BNIP3 Pathway Induced by Circulation Massage
- VernacularTitle:循经按摩通过HIF-1α/BNIP3通路对缺血性脑卒中大鼠脑组织线粒体自噬的影响机制
- Author:
Yiding WEI
1
;
Hai HUANG
1
;
Fei HUANG
1
Author Information
- Publication Type:Journal Article
- Keywords: Meridian massage; Ischemic stroke; Mitochondrial autophagy; HIF-1α/BNIP3 signaling pathway; Hippocampus; Neural function
- From: World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(9):2699-2711
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the effects of meridian massage on mitochondrial autophagy in ischemic stroke rats through the hypoxia-inducible factor-1α(HIF-1α)/BCL2/adenovirus E1B 19kDa interacting protein 3(BNIP3)pathway.Methods The ischemic model was made by blocking the right middle cerebral artery(MCA)of rats with thread embolism.The ischemic stroke rats with successful modeling were divided into model group,massage group,pathway agonist group,pathway inhibitor group and massage+inhibitor group.The rats in the blank control group received routine massage at non-meridian acupoints(including Baihui,Fengfu and Renzhong).The massage group was treated with circulation massage(including Biguan,Taichong,Fenglong and Zusanli),the pathway agonist group was treated with HIF-1α/BNIP3 activator,the pathway inhibitor group was treated with HIF-1α/BNIP3 inhibitor,and the massage+inhibitor group was treated with combination of circulation massage and HIF-1α/BNIP3 inhibitor.Another 10 healthy rats were selected for sham operation intervention,which only separated the blood vessels without inserting the plug.Neurological deficits were evaluated through the Zea Longa scoring system;TTC brain smear staining was used to determine the volume of cerebral infarction and the degree of hemispheric swelling in rats.The pathological changes of rat hippocampal tissue were observed by HE staining;Immunofluorescence was used to detect the expressions of HIF-1α and BNIP3 in rat brain tissue;The number of autophagosomes in the hippocampal region was observed by transmission electron microscopy.Western blot was used to detect the protein expressions related to autophagy(LC3B),apoptosis(Beclin-1),and HIF-1α/BNIP3 signaling pathways in mitochondria of brain tissue.Results ① The fluorescence expression intensity of HIF-1α and BNIP3 in hippocampus of model group was increased compared with that of sham group(P<0.05);The fluorescence expression intensity of HIF-1α and BNIP3 in hippocampus of massage group was significantly increased compared with that of model group(P<0.05).② Compared with model group,cerebral hemisphere swelling,nerve function deficit score and cerebral infarction volume in inhibitor group were increased(P<0.05);The levels of cerebral hemisphere swelling,nerve function deficit score and cerebral infarction volume in agonist group were decreased(P<0.05).The protein expressions of Beclin-1 and LC3B in agonist group were increased(P<0.05).The protein expressions of Beclin-1 and LC3B in inhibitor group were decreased(P<0.05).③ Compared with the massage group,the hippocampal cells in the massage+inhibitor group were more dispersed,the number of cells decreased,edema appeared,and tissue was damaged.④ Compared with model group,the protein expressions of LC3B,HIF-1α,Beclin-1 and BNIP3 in massage group were increased(P<0.05);The protein expressions of LC3B,HIF-1α,Beclin-1 and BNIP3 in massage+inhibitor group were decreased(P<0.05).Conclusion In the treatment of ischemic stroke,meridian massage can activate HIF-1α/BNIP3 signaling pathway,and then promote brain mitochondrial autophagy to improve nerve function,which provides a theoretical basis for the subsequent application of meridian massage in the clinical treatment of ischemic stroke patients.
