基于mTOR/p70S6K信号通路探讨艾灸对阿尔茨海默病小鼠自噬的影响  

Effect of moxibustion on autophagy in mice with Alzheimer’s disease based on mTOR/p70S6K signaling pathway

作　　者：吴洋洋 宋小鸽[2] 朱才丰[3] 蔡圣朝[3] 葛侠[4] 王玲 贾玉梅 WU Yang-yang;SONG Xiao-ge;ZHU Cai-feng;CAI Sheng-chao;GE Xia;WANG Ling;JIA Yit-mei(Graduate School of Anhui University of CM,Hefei 230038,China;Institute of Acupuncture-Moxibustion and Meridians,Anhui University of CM,Second Affiliated Hospital of Anhui University of CM,Hefei 230061;Department of Geriatrics,Second Affiliated Hospital of Anhui University of CM,Hefei 230061;Department of Endocrinology,Second Affiliated Hospital of Anhui University of CM,Hefei 230061)

机构地区：[1]安徽中医药大学研究生院,合肥230038 [2]安徽中医药大学针灸经络研究所,合肥230061 [3]安徽中医药大学第二附属医院老年病科,合肥230061 [4]安徽中医药大学第二附属医院内分泌科,合肥230061

出　　处：《中国针灸》2022年第9期1011-1016,共6页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金资助项目:81603701;安徽省高校自然科学研究项目:KJ2020A0398;国家中医药管理局全国名老中医药专家传承工作室蔡圣朝工作室建设项目:国中医药人教发[2014]20号;安徽省医疗卫生重点专科建设项目:皖卫科教发[2021]273号;安徽省自然科学基金项目:2008085QH391。

摘　　要：目的:探讨艾灸对淀粉样前体蛋白/早老素1(APP/PS1)双转基因阿尔茨海默病(AD)小鼠自噬和β-淀粉样蛋白1-42(Aβ)蛋白表达的影响。方法:将56只6月龄APP/PS1双转基因AD小鼠适应性饲养2个月后,随机分为模型组、艾灸组、雷帕霉素组、抑制剂组,每组14只;14只同月龄C57BL/6J小鼠作为正常组。艾灸组小鼠予隔附子饼实按灸“百会”“风府”“大椎”20 min,雷帕霉素组小鼠予腹腔注射雷帕霉素(2 mg/kg),抑制剂组小鼠在艾灸组基础上注射1.5mg/kg3-甲基腺嘌呤(3-MA),均每日1次,连续2周。HE染色法观察各组小鼠海马组织形态,透射电镜观察各组小鼠海马组织细胞超微结构,免疫组化法检测各组小鼠额叶皮质和海马组织Aβ蛋白表达,Western blot法检测各组小鼠海马组织雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、P70核糖体蛋白S6激酶(p70S6K)和磷酸化p70S6K(p-p70S6K)蛋白表达。结果:与正常组比较,模型组小鼠神经元细胞数量减少,细胞坏死且变形,自噬泡和溶酶体减少。与模型组比较,艾灸组和雷帕霉素组小鼠神经元细胞数量增多,细胞坏死减少,自噬泡和溶酶体增多。与正常组比较,模型组小鼠Aβ、mTOR、p-mTOR、p70S6K、p-p70S6K蛋白表达增多(P<0.05);与模型组比较,艾灸组、雷帕霉素组和抑制剂组小鼠Aβ、mTOR、p-mTOR、p70S6K、p-p70S6K蛋白表达减少(P<0.05);与抑制剂组比较,艾灸组、雷帕霉素组小鼠Aβ、mTOR、p-mTOR、p70S6K、p-p70S6K蛋白表达减少(P<0.05);与雷帕霉素组比较,艾灸组小鼠mTOR、p-mTOR、p70S6K、p-p70S6K蛋白表达减少(P<0.05)。结论:艾灸可增强APP/PS1双转基因AD小鼠海马组织细胞自噬,减少脑组织Aβ异常聚集,其机制可能与抑制m TOR/p70S6K信号通路有关。Objective To investigate the effect of moxibustion on autophagy and amyloidβ-peptide1-42(Aβ)protein expression in amyloid precursor protein/presenilin 1(APP/PS1)double-transgenic mice with Alzheimer’s disease(AD).Methods After 2-month adaptive feeding,fifty-six 6-month-old APP/PS1 double transgenic AD mice were randomly divided into a model group,a moxibustion group,a rapamycin group and an inhibitor group,14 mice in each group.Another 14C57BL/6J mice with the same age were used as a normal group.The mice in the moxibustion group were treated with monkshood cake-separated moxibustion at"Baihui"(GV 20),"Fengfu"(GV 16)and"Dazhui"(GV 14)for 20 min;the mice in the rapamycin group were intraperitoneally injected with rapamycin(2 mg/kg);the mice in the inhibitor group were treated with moxibustion and injection of 1.5 mg/kg 3-methyladenine(3-MA).All the treatments were given once a day for consecutive 2 weeks.The morphology of hippocampal tissue was observed by HE staining;the ultrastructure of hippocampal tissue was observed by transmission electron microscopy;the expression of Aβprotein in frontal cortex and hippocampal tissue was detected by immunohistochemistry;the expressions of mammalian target of rapamycin(mTOR),phosphorylated mTOR(p-m TOR),p70 ribosomal protein S6 kinase(p70S6K)and phosphorylated p70S6K(p-p70S6K)protein in hippocampus were detected by Western blot method.Results Compared with the normal group,the number of neuron cells was decreased,cells were necrotic and deformed,and autophagy vesicle and lysosome were decreased in the model group.Compared with the model group,the number of neuron cells was increased,cell necrosis was decreased,and autophagy vesicle and lysosome were increased in the moxibustion group and the rapamycin group.Compared with the normal group,the protein expressions of Aβ,mTOR,p-m TOR,p70S6K and p-p70S6K in the model group were increased(P<0.05);compared with the model group,the protein expressions of Aβ,mTOR,p-m TOR,p70S6K and p-p70S6K in the moxibustion group,rapamycin g

关 键 词：阿尔兹海默病 艾灸 APP/PS1双转基因AD小鼠 Β-淀粉样蛋白 细胞自噬 mTOR/p70S6K信号通路 

分 类 号：R245.81[医药卫生—针灸推拿学]