电针通过蛋白激酶B和雷帕霉素机制靶点信号通路抑制睡眠不足大鼠的海马氧化应激和自噬作用

Zheng Peng, Meng Ying, Liu Meijun, Y U Di, Liu Huiying, Wang Fuchun, X U Xiaohong
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引用次数: 0

摘要

目的研究针灸对睡眠剥夺诱导的大鼠学习记忆障碍、氧化应激和自噬的影响,并分析其相关机制:方法:将 30 只 Wistar 大鼠随机分为正常组、睡眠剥夺组和针刺组。采用改良的多平台睡眠剥夺法建立大鼠睡眠剥夺模型。大鼠的百会穴(GV20)、神门穴(HT7)和三阴交穴(SP6)定位电针(密度波,频率20 Hz,强度1 mA),保持针感,留针15 min,连续针刺7 d。用检测试剂盒检测大鼠脑内丙二醛(MDA)的含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPX)的活性,以及自噬相关蛋白轻链3α(LC3A)、轻链3β(LC3B)和轻链3β(LC3C)的含量、轻链 3 beta(LC3B)和 Beclin 1,以及蛋白激酶 B(PKB/AKT)和雷帕霉素机械靶标(mTOR)信号通路的激活情况。结果显示与正常组相比,睡眠剥夺组大鼠在目标象限停留的时间(P 0.05)和进入目标象限的次数(P 0.05)明显减少,MDA含量明显升高(P 0.01),脑内SOD和GPX活性(P 0.01)明显降低,LC3A Ⅱ/Ⅰ、LC3B Ⅱ/Ⅰ和Beclin 1明显升高(P 0.01),p-AKT(ser473)/AKT、p-mTOR(ser2448)/mTOR和p-p70s6K(thr389)/p70S6明显降低(P 0.01)。与睡眠剥夺组相比,针刺组大鼠治疗7 d后在目标象限停留的时间和进入目标象限的次数(P 0.05)均明显增加,MDA含量明显降低(P 0.05),脑内SOD和GPX活性明显增加(P 0.05)。此外,LC3A Ⅱ/Ⅰ、LC3BⅡ/Ⅰ和Beclin 1的水平明显下降(P 0.05),p-AKT(ser473)/AKT、p-mTOR(ser2448)/mTOR和p-p70s6K(thr389)/p70s6k的水平明显升高(P 0.05):结论:针灸能明显改善睡眠剥夺导致的学习和记忆损伤,抑制氧化应激和自噬,其作用与AKT/mTOR信号的激活有关。
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Electroacupuncture inhibits hippocampal oxidative stress and autophagy in sleep-deprived rats through the protein kinase B and mechanistic target of rapamycin signaling pathway.

Objective: To investigate the effects of acupuncture on learning and memory impairment, oxidative stress and autophagy induced by sleep depriv ation in rats, and to analyze the related mechanism.

Methods: Thirty Wistar rats were randomly divided into a normal group, sleep deprivation group and acupuncture group. The rat model of sleep deprivation was established by a modified multiplatform sleep deprivation method. The Baihui (GV20), Shenmen (HT7) and Sanyinjiao (SP6) acupoints of rats were located to give electroacupuncture (density wave, frequency 20 Hz, intensity 1 mA) to maintain the needle feeling, and to keep the needle for 15 min and continuous acupuncture for 7 d. The spatial learning and memory abilities of the rats were detected by the water maze test. The content of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) in the brain were detected by an assay kit, and the autophagy related proteins light chain 3 alpha (LC3A), light chain 3 beta (LC3B) and Beclin 1 and the activation of the protein kinase B (PKB/AKT) and mechanistic target of rapamycin (mTOR) signaling pathway in the rat's brain were detected by Western blotting.

Results: Compared with the normal group, the time spent in the target quadrant (P < 0.05) and the number of times entering the target quadrant (P < 0.05) in the rats of sleep deprivation group were significantly reduced, and the content of MDA was significantly increased (P < 0.01), while the activities of SOD and GPX (P < 0.01) in the brain were significantly decreased, and LC3A Ⅱ/Ⅰ, LC3B Ⅱ/Ⅰ and Beclin 1 increased significantly (P < 0.01), while p-AKT (ser473)/AKT, p-mTOR (ser2448)/mTOR and p-p70s6K (thr389)/p70S6 decreased significantly (P < 0.01). Compared with the sleep deprivation group, the time spent in the target quadrant and the times of entering the target quadrant (P < 0.05) in the rats of acupuncture group after 7 d of treatment were significantly increased, Additionally, the content of MDA was significantly decreased (P < 0.05), while the activities of SOD and GPX (P < 0.05) in the brain were significantly increased. Moreover, the levels of LC3A Ⅱ/Ⅰ, LC3BⅡ/Ⅰ and Beclin 1 decreased significantly (P < 0.05), and that of p-AKT (ser473)/AKT, p-mTOR (ser2448)/mTOR and p-p70s6K (thr389)/p70s6k increased significantly (P < 0.05).

Conclusion: Acupuncture can significantly improve the learning and memory damage caused by sleep deprivation and inhibit oxidative stress and autophagy, and its effect is related to the activation of AKT/mTOR signaling.

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