基于Nrf2/ROS通路分析醒脑开窍针刺对脑出血偏瘫大鼠神经功能与痉挛状态的影响

    Study on the effects of Xingnao Kaiqiao acupuncture on the neurological function and spastic state of hemiplegic rats with intra cerebral hemorrhage based on the Nrf2/ROS pathway

    • 摘要: 目的: 探讨醒脑开窍针刺(XNKA)对脑出血(ICH)模型大鼠神经功能恢复的作用及基于Nrf2信号通路的抗氧化应激机制。方法: 将40只SD大鼠随机分为假手术组(Sham)、模型组(Model)、XNKA组和假针刺组(Sham-Acup)。采用Rosenberg法建立ICH模型,XNKA组于造模24 h后针刺“人中、内关、三阴交”等穴,持续干预14 d。通过改良神经功能缺损评分(mNSS)、改良Ashworth量表(MAS)及运动诱发电位(MEP)评估神经功能;HE与TUNEL染色观察脑组织病理及凋亡;Western blotting检测Ke1ch样环氧氯丙烷相关蛋白-1(Keap1)、核因子E2相关因子2(Nrf2)和血红素加氧酶1(HO-1)蛋白表达;试剂盒测定超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)及活性氧(ROS)水平。结果: 与Model组相比,XNKA组神经功能显著改善(P < 0.05),痉挛状态减轻(P < 0.05),MEP潜伏期缩短(P < 0.05)。病理结果显示XNKA组神经元损伤及凋亡减少,同时下调Keap1并激活Nrf2/HO-1信号通路(P < 0.05),显著提升SOD与GSH-Px活性(P < 0.05),降低MDA及ROS水平(P < 0.05)。结论: XNKA通过激活Nrf2/HO-1信号通路,增强抗氧化酶活性, 抑制氧化应激与神经元凋亡,从而促进ICH后神经功能恢复。

       

      Abstract: Objective: To investigate the effects of Xingnao Kaiqiao acupuncture (XNKA) on the recovery of neurological function in rats with intracerebral hemorrhage(ICH), and antioxidant stress mechanism based on the Nrf2 signaling pathway. Methods: Forty SD rats were randomly divided into the sham operation group(Sham), model group(Model), XNKA group and sham acupuncture group(Sham-Acup).The ICH model was established by the Rosenberg method, and the XNKA group were treated with "Renzhong, neiguan, and sanyinjiao" acupuncture after 24 h of modeling for 14 days.The neurological function was evaluated by the modified neurological deficit score(mNSS), modified ashworth scale(MAS) and motor evoked potential(MEP).HE and TUNEL staining were used to observe the pathology and apoptosis of brain tissue.The protein expressions of Keap1, Nrf2 and HO-1 were detected by Western blotting.The levels of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), malondialdehyde(MDA) and reactive oxygen species(ROS) were determined by the kit. Results: Compared with the Model group, the neurological function was significantly improved(P < 0.05), the spastic state was alleviated(P < 0.05), and the MEP latency was shortened in the XNKA group(P < 0.05).The pathological results showed that the neuronal injury and apoptosis in the XNKA group were reduced.At the same time, the Keap1 was down-regulated, the Nrf2/HO-1 signaling pathway was activated(P < 0.05), the activities of SOD and GSH-Px were significantly increased(P < 0.05), and the levels of MDA and ROS were decreased(P < 0.05). Conclusions: XNKA promotes the recovery of neurological function after ICH by activating the Nrf2/HO-1 signaling pathway, enhancing the activity of antioxidant enzymes and inhibiting oxidative stress and neuronal apoptosis.

       

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