Effect of Moxibustion of Governor Vessel on Autophagy Level of APP/PS1 Double-Transgenic Mice by Regulating Wnt/β-catenin Signaling Pathway
Objective:To explore the effect of moxibustion of governor vessel on the related protein expressions of autophagy and Wnt/β-catenin signaling pathway in amyloid precursor protein/presenilin 1(APP/PS1)double transgenic mice to provide experimental basis for the prevention and treatment of Alzheimer's disease.Methods:Total twelve C57BL/6J male mice were set as the normal group,and 48 APP/PS1 double transgenic mice of the same age and background were randomly divided into the model group,moxibustion group,rapamycin group and moxibustion+inhibitor group with 12 mice in each group.Aconite cake-separated moxibustion at Baihui(DU20)and mild moxibustion at Fengfu(DU16)and Dazhui(DU14)were applied in the moxibustion group.Rapamycin group was intraperitoneally injected with 2 mg/kg rapamycin,moxibustion+inhibitor group on the basis of moxibustion group,interitoneally injected 3-methyladenine 1.5 mg/kg,and all the three groups were intervened once a day for 2 weeks and stopped once a week.Mice in the normal group and model group were fixed without any intervention.Water maze test was performed before and after intervention to detect escape latency.The number and morphology of autophagosomes and autophagy vesicles in the hippocampus of mice were observed by transmission electron microscope,and the protein expression of β-amyloid 1-42(Aβ1-42)was detected by immunohistochemistry.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)technology was used to detect the relative expression levels of Aβ1-42 mRNA in the hippocampal region of the brain.Western blot(WB)analysis was utilized to examine the protein expression of microtubule-associated protein light chain 3 Ⅱ(LC3 11)/microtubule-associated protein light chain 3 Ⅰ(LC3 Ⅰ),ubiquitin-binding protein 62(p62),rapamycin target protein(mTOR),Wnt family member 3a(Wnt3a),glycogen synthase kinase-3 β(GSK3 β),and β-catenin in the hippocampal tissue.Results:After intervention,compared with the model group,the escape latency of moxibustion group and rapamycin group were shortened(P＜0.05).There were fewer autophagy vesicles and autophagosomes in the hippocampus of the model group and moxibustion+inhibitor group,and more autophagy vesicles and autophagosomes in the moxibustion group and rapamycin group.Compared with the normal group,the particle number of positive expression of Aβ1-42 in the model group increased(P＜0.05),while the particle number of positive expression of Aβ1-42 in the moxibustion group and rapamycin group decreased compared with the model group(P＜0.05).Compared with the normal group,the relative expression of Aβ142 mRNA in the model group increased(P＜0.05).The relative expression of Aβ1_42 mRNA in the moxibustion group and rapamycin group decreased compared with the model group(P＜0.05).Compared with the normal group,the protein expressions of p62,mTOR and GSK-3β in the model group increased(P＜0.05),while the protein expressions of LC3Ⅱ/LC3 Ⅰ,Wnt3a and β-catenin in the model group decreased(P＜0.05).Compared with the model group,the protein expressions of p62,mTOR and GSK-3β in the moxibustion group and rapamycin group decreased(P＜0.05),while the protein expressions of LC3 11/LC3 Ⅰ,Wnt3a and β-catenin increased(P＜0.05).Compared with the moxibustion+inhibitor group,the protein expressions of p62,mTOR and GSK-3βin the moxibustion group and rapamycin group decreased(P＜0.05),while the protein expressions of LC3 Ⅱ/LC3 Ⅰ,Wnt3a and β-catenin in the moxibustion group and rapamycin group increased(P＜0.05).Conclusions:Moxibustion of governor vessel can accelerate the clearance of Aβ in the brain of APP/PS1 double transgenic mice,the mechanism of which may be related to the activation of Wnt signaling pathway by moxibustion of governor vessel,and the promotion of autophagy.
万方数据
维普
