电针可减轻脊髓损伤后HIF1-α介导的早期线粒体自噬
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浙江中医药大学第三临床医学院-浙江省针灸与神经病学重点实验室,杭州310053,浙江,中国

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Electroacupuncture Alleviates HIF1-α-mediated Early Mitophagy in Spinal Cord Injury
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Key Laboratory of Acupuncture and Neurology of Zhejiang Province, The Third School of Clinical Medicine (School of Rehabilitation Medicine), Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China

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    摘要:

    背景: 脊髓损伤周围的抑制性微环境严重限制了损伤后的功能修复。线粒体自噬是维持细胞内稳态、保证神经细胞微环境平衡的重要措施之一。缺氧诱导因子1α (hypoxia inducible factor1α,HIF1-α)可介导神经退行性疾病的线粒体自噬,但其机制复杂多样,有待进一步阐明。电针在改善脊髓损伤后神经微环境,促进脊髓损伤患者长期神经功能恢复方面具有显著作用,但电针是否参与HIF1-α介导的线粒体自噬尚不清楚。目的: 探讨HIF1-α对脊髓损伤大鼠线粒体自噬的影响及电针作用的可能机制。方法: 采用改良Allen法构建SD大鼠SCI模型后,电针干预T9、T11夹脊穴,进一步进行分子生物学和形态学研究。通过立体定位注射HIF1-α敲低病毒,通过免疫印迹、实时荧光定量PCR、免疫荧光、透射电镜和尼氏染色检测损伤局部区域线粒体自噬的变化,观察HIF1-α对局部损伤修复的影响。结果: HIF1-α及其线粒体自噬受体BNIP3和NIX在脊髓损伤后表达上调。电针治疗或局部抑制HIF1-α表达可逆转脊髓损伤后早期自噬状态,减少细胞凋亡和损伤面积,促进神经元存活。结论: 电针可能通过减轻HIF1-α介导的早期线粒体自噬成为一种治疗脊髓损伤的潜在策略。

    Abstract:

    Background The inhibitory microenvironment around spinal cord injury (SCI) severely restricted functional repair after injury. Mitophagy was one of the important measures to maintain cellular homeostasis and ensure the harmonious nerve cell microenvironment. Hypoxia-inducible factor1α (HIF1-α) can mediate mitochondrial autophagy in neurodegenerative diseases, but the mechanisms are complex and diverse, which need to be further elucidated. Electroacupuncture plays a significant role in improving the neural microenvironment after spinal cord injury, promote long-term neurological function recovery in SCI patients, but whether electroacupuncture can participate in HIF1-α mediated mitophagy remains unknown. Objective Investigated the effects of HIF1-α on mitochondrial autophagy in rats with spinal cord contusion and the potential mechanism of electroacupuncture. Methods Following the successful construction of an SCI model of Sprague-Dawley rat utilizing a modified Allen method, electroacupuncture intervention was performed at T9 and T11 Jiaji acupoint (EX-B2), with further molecular biology and morphology examined by perfusion. To observe the effect of HIF1-α on local damage repair, the stereotypic injection of Hif1a knockdown virus was performed, and the changes of mitophagy in damaged local area was detected employing Western blotting, real-time fluorescence quantitative PCR, immunofluorescence, transmission electron microscopy and Nissl staining. Results HIF1-α as well as its mitophagy receptor BNIP3 and NIX are upregulated after spinal cord injury. Electroacupuncture treatment or local inhibition of HIF1-α expression can reverse the early autophagy state after spinal cord injury, reduce cell apoptosis and injury area, promote neuronal survival. Conclusion Electroacupuncture may serve as a promising strategy for spinal cord injury treatment, by alleviating HIF1-α mediated early mitochondrial autophagy.

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  • 在线发布日期: 2024-04-15
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