Ma Qin, Shao Rui-Jie, Wei Gao-Wen, Lu Jun, Wu Ji-Hong, Tu Ya
School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 100029, China.
Zhen Ci Yan Jiu. 2024 Oct 25;49(10):1063-1069. doi: 10.13702/j.1000-0607.20230726.
To observe the effect of manual acupuncture stimulation on changes of behavior and hippocampal signaling pathways of AMP-activated kinase (AMPK)/UNC-51-like kinase 1 (ULK1) in rats with chronic unpredictable mild stress (CUMS)-induced depression, so as to explore its molecular mechanism underlying improvement of depression.
Thirty-two male SD rats were randomly divided into normal control, model, acupuncture and medication (fluoxetine) groups, with 8 rats in each group. The depression model was established by using CUMS (fasting, water depredation, restraint, swimming in cold water, crowding, stroboscope stimulation, and tail clipping, each of which was used once a week) for 6 weeks. Manual acupuncture stimulation was applied to "Baihui"(GV20) and "Yintang"(EX-HN3) for 10 min, once daily, 6 times a week for 6 weeks before every-day modeling. Rats of the medication group received gavage of fluoxetine (10 mg/kg) once a day for 6 weeks before every-day modeling. The percentage of sucrose preference, and the percentages of open arm times and open arm time in the elevated plus maze experiments were detected. The expression levels of AMPK, phosphorylated (p)-AMPK, ULK1, p-ULK1, mammalian target of rapamycin (mTOR) and p-mTOR proteins in the hippocampus tissue were detected by Western blot, and the expression of autophagy effecting protein (Beclin-1) and the selective autophagy receptors (p62) mRNA in the hippocampal tissue was detected using real-time fluorescence quantitative PCR, respectively.
Compared with the normal group, the percentage of sucrose preference, percentage of times of entering the open arm and percentage of open arm time, the expression levels of p-AMPK /AMPK ratio, p-ULK1 /ULK1 ratio of proteins, and Beclin-1 mRNA were significantly decreased (<0.01, <0.05), and the expression levels of p-mTOR /mTOR ratio and p62 mRNA were significantly increased (<0.01, <0.05) in the model group. In comparison with the model group, the percentage of sucrose preference, percentage of open arm times and open arm time, the expression levels of p-AMPK /AMPK ratio, p-ULK1/ULK1 ratio of proteins, and Beclin-1 mRNA were significantly increased in both acupuncture and medication groups (<0.01, <0.05), while the expression levels of p-mTOR /mTOR ratio of proteins, and p62 mRNA were markedly decreased only in the medication group (<0.05).
Acupuncture treatment can alleviate depression-like behavior in CUMS rats, which may be related to its functions in up-regulating AMPK/ULK1 signaling pathway to induce cellular autophagy in the hippocampus.
观察手针刺激对慢性不可预测轻度应激（CUMS）诱导的抑郁大鼠行为及海马中AMP激活的蛋白激酶（AMPK）/UNC-51样激酶1（ULK1）信号通路变化的影响，以探讨其改善抑郁的分子机制。
将32只雄性SD大鼠随机分为正常对照组、模型组、针刺组和药物（氟西汀）组，每组8只。采用CUMS（禁食、禁水、束缚、冷水游泳、拥挤、频闪灯刺激和夹尾，每周各进行1次）建立抑郁模型6周。在每天造模前，针刺组针刺“百会”（GV20）和“印堂”（EX-HN3）10分钟，每日1次，每周6次，共6周。药物组在每天造模前灌胃给予氟西汀（10 mg/kg），每日1次，共6周。检测蔗糖偏好百分比、高架十字迷宫实验中进入开放臂次数百分比和开放臂时间百分比。采用蛋白质免疫印迹法检测海马组织中AMPK、磷酸化（p）-AMPK、ULK1、p-ULK1、哺乳动物雷帕霉素靶蛋白（mTOR）和p-mTOR蛋白的表达水平，采用实时荧光定量PCR法检测海马组织中自噬效应蛋白（Beclin-1）和选择性自噬受体（p62）mRNA的表达。
与正常组相比，模型组蔗糖偏好百分比、进入开放臂次数百分比和开放臂时间百分比、蛋白p-AMPK /AMPK比值、p-ULK1 /ULK1比值以及Beclin-1 mRNA表达水平均显著降低（<0.01，<0.05），而蛋白p-mTOR /mTOR比值和p62 mRNA表达水平显著升高（<0.01，<0.05）。与模型组相比，针刺组和药物组的蔗糖偏好百分比、开放臂次数百分比和开放臂时间、蛋白p-AMPK /AMPK比值、p-ULK1/ULK1比值以及Beclin-1 mRNA表达水平均显著升高（<0.01，<0.05），而仅药物组蛋白p-mTOR /mTOR比值和p62 mRNA表达水平显著降低（<0.05）。
针刺治疗可减轻CUMS大鼠的抑郁样行为，这可能与其上调AMPK/ULK1信号通路以诱导海马细胞自噬的功能有关。
