针刺对慢性应激抑郁大鼠海马AMPK/ULK1信号通路和自噬的影响  

Effect of acupuncture on AMPK/ULK1 signaling pathway of hippocampus in chronic stressinduced depression rats

作　　者：马琴 邵瑞洁 魏高文 卢峻[1] 邬继红[1] 图娅[1] 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)

机构地区：[1]北京中医药大学针灸推拿学院,北京100029

出　　处：《针刺研究》2024年第10期1063-1069,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.82174508)。

摘　　要：目的:观察针刺对慢性应激抑郁大鼠行为学及海马腺苷酸活化蛋白激酶(AMPK)/UNC-51样激酶1(ULK1)信号通路的影响,探讨针刺治疗抑郁症的分子机制。方法:将32只雄性SD大鼠随机分成正常组、模型组、针刺组和氟西汀组,每组8只。采用慢性不可预知温和应激方法制备抑郁大鼠模型,共6周。针刺组每天造模前针刺“百会”和“印堂”,每次10 min,每周6次;氟西汀组每天造模前以氟西汀(10 mg/kg)灌胃,每日1次;以上干预均持续6周。检测大鼠糖水偏好率、高架十字迷宫实验开臂次数百分比和开臂时间百分比;采用Western blot法检测海马组织AMPK、磷酸化(p)-AMPK、ULK1、p-ULK1、哺乳动物类雷帕霉素靶蛋白(mTOR)、p-mTOR蛋白表达;采用实时荧光定量PCR法检测海马组织自噬效应蛋白Beclin-1、选择性自噬受体p62 mRNA表达。结果:与正常组比较,模型组大鼠糖水偏好率、进入开臂次数百分比和开臂时间百分比均降低(P<0.01,P<0.05),海马p-AMPK/AMPK、p-ULK1/ULK1蛋白表达比值及Beclin-1mRNA表达水平下降(P<0.01,P<0.05),海马p-mTOR/mTOR蛋白表达比值和p62 mRNA表达水平升高(P<0.01,P<0.05)。与模型组比较,针刺组和氟西汀组大鼠糖水偏好率、进入开臂次数百分比和开臂时间百分比均升高(P<0.01,P<0.05),海马p-AMPK/AMPK、p-ULK1/ULK1蛋白表达比值及Beclin-1 mRNA表达水平升高(P<0.01,P<0.05);氟西汀组大鼠海马p-mTOR/mTOR蛋白表达比值及p62 mRNA表达水平降低(P<0.05)。结论:针刺可能通过激活海马AMPK/ULK1通路、促进自噬,改善慢性应激大鼠抑郁样行为。Objective 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.Methods 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.Results 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(P<0.01,P<0.05),and the expression levels of p-mTOR/mTOR ratio and p62 mRNA were significantly increased(P<0.01,P<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

关 键 词：针刺 抑郁症 慢性应激 腺苷酸活化蛋白激酶/UNC-51样激酶1信号通路 自噬 

分 类 号：R245[医药卫生—针灸推拿学]