Study On The Effects Of Electroacupuncture On The Levels Of Hippocampal LC3, CTSD And A? In APP/PS1 Double Transgenic Mice At The Age Of 4 Months

In recent years,with the gradual aging of society,Alzheimer's disease(AD)has also received more and more attention.In the past 20 years,the amyloid(A?)peptide cascade theory has been the core of research,treatment and development of Alzheimer's disease,but the drugs based on this hypothesis have not successfully entered the market.Although the failure of clinical trials of such drugs has cast a shadow over the A? cascade theory,there is still much evidence that A? is at the heart of the disease development processNow,lysosomal function and abnormalities in autophagy are also considered to be hallmarks of Alzheimer's disease(AD),and lysosomal and autophagic dysfunction are closely related to the formation and decomposition of ?-amyloid.Usually,during the process of lysosomal degradation,the recovered waste cell components and damaged organelles are transported to the lysosomes mainly by transport of endosomes and autophagosomesWith the continuous advancement and development of modern research,the treatment of AD has become more and more diversified.Because the pathogenesis of AD is extremely complicated in the late stage of its procedure,and needs to be further clarified,the timing of treatment for AD more and more inclined to the early stages.In the research of acupuncture and moxibustion in Chinese medicine,it shows the therapeutic effect of acupuncture on AD,and treating in advance has always been advantage of the acupuncture and moxibustion of Chinese medicine.In the previous experiments of this group,electroacupuncture intervention in AD transgenic mice can improve their cognitive dysfunction and reduce the level of A? in their brain.Therefore,in this experiment,we selected 4 months old AD transgenic mice to perform electroacupuncture intervention,observe the effect of electroacupuncture intervention on the early pathological state of AD transgenic mice,and discuss the mechanism of electroacupuncture intervention on AD from the perspective of lysosomes and autophagyObjectiveInvestigating the effects of electroacupuncture on LC3,cathepsin D(CTSD)and A?,amyloid precursor protein(APP)in hippocampus of 4 month old APP/PS1 double transgenic mice and their relationship.Discussing the mechanism of EA on AD from the perspective of lysosomes and autophagy.MethodsA total of 16 male APP/PS1 transgenic mice were divided into electroacupuncture group and model group randomly.There were 8 male C57BL/6 mice selected as normal control group.The expression of LC3 in brain was observed by immunohistochemistry after 6 weeks of electroacupuncture at Baihui(DU 20)and Yongquan(KI 1).Co-expression of LC3 and A?,CTSD and A? in brain was observed by immunofluorescence double labeling.The level of LC3 ?,LC3?,CTSD and APP in hippocampus was detected by Western blot(WB)ResultsImmunohistochemistry results showed that expression of LC3 was in neurons in hippocampus and cortex.The results of immun of luorescence double labeling showed that there was co-expression of LC3 and A?,CTSD and A?.WB results showed that the level of LC3 ?/LC3 I in the model group(P<0.01)and electroacupuncture group(P<0.05)both were lower than the level in control group.Compared with the model group,the average value of LC3?/LC3 I in the electroacupuncture group was increased(P>0.05,P=0.19).The level of CTSD in the model group was lower than the level in control group(P<0.01)and electroacupuncture group(P<0.05).The level of APP in the control group was lower than the level in model group(P<0.01),and compared with the model group,the level of APP in the electroacupuncture group was decreased(P>0.05).Correlation analysis results,the level of LC3?/LC3I in hippocampus of mice was negatively correlated with the level of APP(r=-0.563,P<0.01).The content of CTSD in hippocampus of mice was negatively correlated with the level of APP(r=-0.50,P<0.05).The content of CTSD in hippocampus of mice was positively correlated with the level of LC3(r=0.53,P<0.01)ConclusionAt the age of 5 months in APP/PS1 double transgenic mice,autophagy in the hippocampus was inhibited and lysosomal degradation was impaired.Electroacupuncture intervention for 4 months old APP/PS1 double transgenic mice can increase the expression level of CTSD.Electroacupuncture may reduce the level of A? in the brain by enhancing the lysosomal and autophagic lysosomal degradation functions in hippocampus.The effect of electricity on autophagy function needs to be confirmed by further experiments.