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

doi:10.13703/j.0255-2930.20210705-k0004

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

更新时间：2023-05-10

- Effect of moxibustion on autophagy in mice with Alzheimer's disease based on mTOR/p70S6K signaling pathway
- Chinese Acupuncture and Moxibustion Vol. 42, Issue 9, Pages: 1011-1016(2022)
- 作者机构：
  
  1. 安徽中医药大学研究生院
  2. 安徽中医药大学针灸经络研究所
  3. 安徽中医药大学第二附属医院老年病科
  4. 安徽中医药大学第二附属医院内分泌科
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20210705-k0004
  CLC： R245
- Published：2022
- 稿件说明：
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Effect of moxibustion on autophagy in mice with Alzheimer's disease based on mTOR/p70S6K signaling pathway[J]. Chinese Acupuncture and Moxibustion, 2022, 42(9): 1011-1016.
DOI：

Effect of moxibustion on autophagy in mice with Alzheimer's disease based on mTOR/p70S6K signaling pathway[J]. Chinese Acupuncture and Moxibustion, 2022, 42(9): 1011-1016. DOI： 10.13703/j.0255-2930.20210705-k0004.

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_(1-42))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_(1-42)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_(1-42)、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_(1-42)、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_(1-42)、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。

Abstract

Objective To investigate the effect of moxibustion on autophagy and amyloid β-peptide1-42(Aβ

1-42

) 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 Aβ

1-42

protein 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

1-42

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

1-42

mTOR

p-m TOR

p70S6K and p-p70S6K in the moxibustion group

rapamycin group and inhibitor group were decreased（P

0.05）;compared with the inhibitor group

the protein expressions of Aβ

1-42

mTOR

p-m TOR

p70S6K and p-p70S6K in the moxibustion group and rapamycin group were decreased（P

0.05）;compared with the rapamycin group

the protein expressions of mTOR

p-m TOR

p70S6K and p-p70S6K in the moxibustion group were decreased（P

0.05）.Conclusion Moxibustion could enhance autophagy in hippocampal tissue of APP/PS1 double transgenic AD mice and reduce abnormal Aβ aggregation in brain tissue

the mechanism may be related to the inhibition of mTOR/p70S6K signaling pathway.

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