The Effect Of Grain-sized Moxibustion On Behavioral Science And GFAP Expression In Alzheimer’s Disease Transgenic Mice

Objective:To observe the effect of grain-sized moxibustion at the point of Xinshu(BL15)and Shenshu(BL23)on learning and memory ability of Alzheimer’s disease transgenic mice.and observed in AD double transgenic mice behavior changes and the expression of GFAP and GFAP and Aβ1-42 plaque co localization,autophagy related protein LC3 expression and so on,to investigate the mechanism of moxibustion treatment of AD.In order to further explain the moxibustion early treatment of AD,to provide experimental evidence for clinical treatment.Methods:The genotyping of transgenic mice were detected by PCR.,40 double transgenic B6SJL-Tg(APPSwFlLon,PSEN1*M146L*L286V)6799Vas/J mice which were 1.5 month old,were randomly divided into treatment group(n=20)and model group(n=20),Normal group adopted the same age and background C57BL/6J female mice(n=20).Xinshu(BL15)and Shenshu(BL23)were cauterized in way of grain-sized moxibustion once a day only in treatment group.The size of moxa cone was 5mm×8mm,Located the moxa cone on Xinshu(BL15)and Shenshu(BL23)and extinguish it when burning to 3/5.The total treatment last for 5.5 months..Have two days rest every ten days treatment.After the treatment,the learning and memory ability of each group was detected by Morris water maze test.After the end of the Morris water maze test in routine pathology,immunofluorescence and immunohistochemistry detection methods were observed in AD transgenic mouse GFAP expression,and GFAP and Aβ1-42 plaque co localization,autophagy related protein LC3 expression and so on.Results:(1)Morris water maze:In the first 6 days of place navigation test,with the increase of the number of training days,the escape latency gradually shortened.The mice in the model group less progress than the treatment group and the normal group and the escape latency was longer,There was a significant difference among three groups(P<0.05);At the seventh day in spatial probe test,compared with the normal group,the times crossed the former location and time staying in the target zone in the model group was significant less(P<0.05),but had no significant difference in treatment group(P>0.05).After moxibustion treatment,the treatment group crossed the former platform location more often than the model group,and time staying in the target zone was longer(P<0.05).(2)Thioflavin-s staining were detected in the brain of amyloid beta deposition results revealed that in the normal group in the brains of mice with no amyloid deposition of positive plaques;compared with model group,moxibustion treatment of mice hippocampus and frontal cortex starch deposition plaque number significantly reduced(P<0.01).(3)Immunohisto chemical method was used to detect the expression of GFAP and LC3 positive cells expression results show:compared with model group mice in frontal cortex and hippocampus of GFAP,LC3 positive cells expression of positive area and the total optical density and normal group,significantly higher,the difference is statistically significant(P<0.01).The group of treatment,compared with model group,treatment group of mice hippocampus GFAP expression levels were significantly decreased(P<0.05)and frontal cortex GFAP expression levels were decreased,and the difference is statistically significant(P<0.01);treatment group of mice hippocampus LC3 expression level decreased,the difference has statistical significance(P<0.05),the positive area of frontal cortex LC3 expression decreased obviously(P<0.05),total optical density values decreased significantly,the difference was significant(P<0.01).Compared with the normal group,there was no significant difference in the expression of GFAP and LC3 positive cells in the hippocampus and frontal cortex of the treatment group(P>0.05).(4)Double immunofluorescence results show:model group AD mouse frontal cortex and hippocampal GFAP and Aβ1-42 positive cells co expression rate was significantly higher than that in the normal group,difference was statistically significant(P<0.05);compared with the model group,treatment group of mice with AD frontal cortex and hippocampal GFAP and amyloid beta 1-42 positive cells co expression rate decreased after treatment,the difference is statistically significant(P<0.05).Conclusion:(1)The early moxibustion therapy can improve the learning and memory ability of AD transgenic mice;(2)Early AD with moxibustion treatment can restrain the expression of AD transgenic mice in frontal cortex and hippocampus of GFAP,amyloid beta 1-42 and LC3,hinder abnormal accumulation of amyloid beta,thereby inhibiting the deposition of amyloid beta;(3)Early AD with moxibustion treatment can inhibit AD transgenic mouse frontal cortex and hippocampus astrocytes excessive proliferation;(4)The mechanism moxibustion effective on AD may realized by restraining the excessive accumulation of β-amyloid protein in the brain of Alzheimer’s disease transgenic mice.