目的 观察艾灸对阿尔茨海默病(Alzheimer's disease,AD)大鼠腺苷酸活化蛋白激酶(adenosine 5-mono-phosphate activated protein kinase,AMPK)/雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路相关递质表达的影响,探讨艾灸治疗AD的作用机制.方法 将SD大鼠按照随机数字表法分为正常组8只、模型组32只,采取侧脑室注射β淀粉样蛋白(amyloid β-protein,Aβ)25-35的方法建立大鼠AD模型.将模型复制成功的大鼠随机分为模型组、药物组、艾灸组,每组8只.艾灸组大鼠用艾条灸"百会""肾俞""三阴交",每次15 min,同时按3 mg/kg灌胃蒸馏水;药物组大鼠按3 mg/kg灌胃盐酸多奈哌齐;对照组及模型组大鼠按3 mg/kg灌胃蒸馏水.采用Morris水迷宫法检测大鼠行为学表现,苏木精—伊红染色法观察大鼠海马病理组织改变,Western blot法检测大鼠海马磷酸化雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin,p-mTOR)、核糖体蛋白S6激酶(ribosomal protein S6 kinase p70,P70S6K)、自噬相关基因5(autophagy-related gene 5,ATG5)、磷酸化腺苷酸活化蛋白激酶(phosphorylated adenosine 5-monophosphate activated protein kinase,p-AMPK)、微管相关蛋白1轻链3B(microtubule associated protein light chain 3B,LC3B)-Ⅱ/LC3B-Ⅰ的表达水平.结果 苏木精—伊红染色结果表明,模型组海马神经元萎缩明显,与模型组比较,药物组和艾灸组海马神经元形态及分化程度均有明显改善.与正常组比较,模型组大鼠的逃避潜伏期显著延长(P<0.05),p-mTOR及P70S6K表达水平均显著升高(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著降低(P<0.05).与模型组比较,药物组和艾灸组大鼠的逃避潜伏期均显著缩短(P<0.05),p-mTOR及P70S6K表达水平均显著下降(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著上升(P<0.05).与药物组比较,艾灸组大鼠逃避潜伏期显著缩短(P<0.05);p-mTOR及P70S6K表达水平显著降低(P<0.05),ATG5、LC3B-Ⅱ/LC3B-Ⅰ、p-AMPK表达水平均显著上升(P<0.05).结论 艾灸能够调控AMPK/mTOR信号通路,诱导细胞自噬,阻断脑内Aβ表达,从而改善认知功能.
Effect of Moxibustion on the Adenosine 5-Monophosphate Activated Protein Kinase/Mammalian Target of Rapamycin Signaling Pathway in the Hippocampus of Rats with Alzheimer's Disease:A Study Based on the Theory of Mutual Help Between the Kidney and the Brain
Objective To investigate the effect of moxibustion on the expression of mediators associated with the adenosine 5-monophosphate activated protein kinase (AMPK )/mammalian target of rapamycin (mTOR )signaling pathway in rats with Alzheimer's disease (AD),as well as the mechanism of action of moxibustion in the treatment of AD.Methods Sprague-Dawley rats were divided into normal group with 8 rats and model group with 32 rats using a random number table.Intracerebroventric-ular injection of β-amyloid 25-35 (Aβ25-35 )was performed to establish a rat model of AD.After modeling,the rats were randomly divided into moxibustion group,model group,and medication group,with 8 rats in each group.The rats in the moxibustion group were given moxa stick moxibustion at Baihui,Shenshu,and Sanyinjiao for 15 minutes each time and 3 mg/kg distilled wa-ter by gavage,those in the medication group were given donepezil hydrochloride at a dose of 3 mg/kg by gavage,and those in the control group and the model group were given 3 mg/kg distilled water by gavage.The Morris water maze test was used to ob-serve the behavioristics of rats;HE staining was used to observe the histopathological changes of the hippocampus;Western blot was used to measure the expression levels of phosphorylated mTOR (p-mTOR ),70-kDa ribosomal protein S6 kinase (P70S6K),autophagy-related gene 5 (ATG5),phosphorylated AMPK (p-AMPK),and microtubule-associated protein light chain 3B- Ⅱ (LC3B- Ⅱ )/microtubule-associated protein light chain 3B- Ⅰ(LC3B- Ⅰ)in the hippocampus of rats.Results HE staining showed that the model group had marked atrophy of hippocampal neurons,and the medication group and the moxibus-tion group had significant improvements in the morphology and differentiation of hippocampal neurons compared with the model group.Compared with the normal group,the model group had a significantly longer escape latency (P<0.05),significant in-creases in the expression levels of p-mTOR and P70S6K (P<0.05),and significant reductions in the expression levels of ATG5,LC3B-Ⅱ/LC3B-Ⅰ,and p-AMPK (P<0.05).Compared with the model group,the medication group and the moxibustion group had a significant reduction in escape latency (P<0.05),significant reductions in the expression levels of p-mTOR and P70S6K (P<0.05),and significant increases in the expression levels of ATG5,LC3B-Ⅱ/LC3B-Ⅰ,and p-AMPK (P<0.05).Compared with the medication group,the moxibustion group had a significant reduction in escape latency (P<0.05),significant reductions in the expression levels of p-mTOR and P70S6K (P<0.05),and significant increases in the expression levels of ATG5,LC3B-Ⅱ/LC3B- Ⅰ,and p-AMPK (P<0.05).Conclusion Moxibustion can improve cognitive function by regulating the AMPK/mTOR signaling pathway,inducing cell autophagy,and blocking the expression of Aβ in brain.
Alzheimer's diseaseMoxibustionAutophagyHippocampusAdenosine 5'-monophosphate-activated protein kinaseMammalian target of rapamycin
万方数据
