Effect Of Moxibustion With Seed-sized Moxa Cone On Aβ Deposition And Neuronal Ultrastructure In The APP/PS1 Transgenic AD Mice

Objective Points of "Xinshu"(BL15)and "Shenshu"(BL23)were slected to treat APP/PS1 transgenic AD mice by moxibustion with seed-sized moxa cone.Observing behavior changes of the AD mice,we investigated the effect of moxibustion with seed-sized moxa cone in the early stage of AD on the expressing levels ofβ-amyloid(Aβ)protein and neuronal ultrastructure.Furthermore,we investigate the possible therapeutic mechanism from above perspective.Methods The genotyping of mice were detected by PCR,APP/PS1 positive transgenic AD mice 40(1.5 month old,18±3g)were divided into 2 groups randomly:20 mice as treatment group,the other 20 as model group.20 negative homozygous mice with the same age and background(wild type)as control group.The treatment group mice were treated with moxibustion with seed-sized moxa cone(Shaped like a cone,5mm × 8mm)on pints of "Xinshu" and "Shenshu".The moxa cone was extinguished and wiped off when it burned to 3/5(in specific case is on the basis of mice’s tolerance).The total treatment lasted for 5.5 months,once a day,10 times for a period with two break days every period.Y maze test performed after treatment.Finally all mice executed to death and got their brain perfused by paraformaldehyde.Applying immunohistochemical method to detect Aβ of hippocampus and cortical,immunofluorescence double labeling to detect co-localization of Aβ1-42 and amyloid fibrils in hippocampus and cortical,TEM to observe the Neuronal Ultrastructure in cortical and hippocampal,ELISA to observe the concentration of Aβ1-40、Aβ1-42 in brain tissue.Results(1)Y maze test:a,Number of electric shocks of mice up to standard:Compared with normal group,model group needed more electric shocks to achieve targets,the difference was significantly(P<0.05),treatment group had no significantly change(P>0.05);Compared with the model group,treatment group had a less electric shocks,the difference was significantly(P<0.05).b,The right memory number tested 24 hours after Y maze test:Compared with normal group,the right memory number of the model group mice was significantly reduced(P<0.05),treatment group had no significantly difference(P>0.05).Compared with the model group,the right memory number of treatment group mice increased.(2)Immunohistochemistry index:Compared with normal group,the positive area of Aβ1-40 and Aβ-42 in frontal cortex and hippocampus of model group increased(P<0.01),total optical density rise too(P<0.01).After moxibustion treatment,compared with the model group,in the hippocampus of treatment group,Aβ1-40 positive and total optical density were decreased significantly(P<0.05),in the frontal cortex of treatment group,Aβ1-40 positive decreased(P<0.01),and total optical density reduced(P<0.05);Aβ1-40 positive area and the total optical density in frontal cortex and hippocampus of treatment group decreased significantly(P<0.01).Compared with the normal group,Aβ1-40,Aβ1-40 positive area and total optical density in frontal cortex and hippocampus of the treatment group expressed no significant difference(P>0.05)(3)Immunofluorescence double labeling index:Compared with normal group,positive cells rate of model group were increased significantly(P<0.01);Compared with the model group,brain positive cells rate of treatment group was significantly reduced(P<0.01);Compared with the normal group,brain positive cells rate of the treatment group had no significant difference(P>0.05).(4)TEM experiment:the mitochondria structure of normal group was not damaged,of model group was badly damaged,of treatment group was slightly damaged;the synapses of normal group was normal,of model group was least,of treatment group was less.(5)ELISA index:Compared with the normal group,the content of Aβ1-40 and Aβ1-42 of model group was significantly increased(P<0.01),contents of Aβ1-40 and Aβ1-42 of treatment group no significant change(P>0.05);Compared with the model group,the content of Aβ1-40 and Aβ1-42 of treatment group was significantly reduced(P<0.01).Conclusion(1)Moxibustion with seed-sized moxa cone have a remarkable effect on improving cognitive function of APP/PS1 transgenic AD mice;(2)Moxibustion with seed-sized moxa cone can improve the neuronal ultrastructure and increase the number of synapses in APP/PS1 transgenic AD mice.(3)Moxibustion with seed-sized moxa cone inhibited the positive expression of Aβ1-40、Aβ1-42 in the hippocampus and frontal cortex of APP/PS1 transgenic AD mice,suggesting that Moxibustion with seed-sized moxa cone had effectively prevention and treatment effect on eraly stage of Alzheimer’s disease.