Orifices Yinao Tang Intervention Role Of Apoptosis, Alzheimer's Disease Model Rat Hippocampal Neurons

Objective:Exploration on pathogenic mechanism of spleen kidney-Qi deficiency and phlegm stasis in Alzheimer's disease by establishing the muti-factor compound rat model of Alzheimer's disease(AD),to explore the mechanism of effect of Qingqiao Yinao Decoction(QYD)in treating AD from neural cells apoptosis and behaviology.Methods:1.The normal 15 months old Wistar rats(n=50)were randomly divided into five groups:normal group,blank group,AD model group,Huperzine A-Zhulin Antun treating group and QYD treating group.There were ten rats in every group.2.To induce AD,five groups rats received injection of IBO andβ-amyloid peptide(Aβ)with BNM through stereospecific instrument.3.After been injectived two weeks,begin to treat them for four weeks,there doses of Qingqiao Yinao Decoction(QYD)were administered by oral gavaga.Huperzine A-Zhulin Autun tablets served as control drug.Normal group,model group and blank group were administered by saline water.4.After treated for four weeks,Morris water maze was used to observing the ability of space learning and memory.There are navigate localizatively experiment and spatial search experiment.To record average incubation period,average swimming distance,the times of pass through the platform,changing tendency of searching the platform.5.After Morris water maze,cut off the rats'heads,taking their brain organization on the ice platform quickly,separates the hippocampus.A part was preservated by beening putted in the liquid nitrogen to test cell apoptosis,another part was fixed by putting them in the 4% paraformmaldehyde,then conventional paraffin wax embedding for immuocytochemistry.6.The protein expression of bcl-2 and bax was examined by using SABC immunocytochemical method.7.Apoptosis level of hippocampus neural cells was determined with flow cytometric analysis. 8.Generalized analysis each examination significance.Results:1.In the behavior experiment,average incubation perriod of Huperzine A-Zhulin Antun treating group and QYD treating group have obvious shorten compared to model group(p＜0.01).The average incubation perriod of Huperzine A-Zhulin Antun treating group and QYD treating group has no difference in the previous four days,the average incubation perriod of QYD treating group has shorten compared to Huperzine A-Zhulin Antun treating group.2.Percentage of quadrant of QYD treating group have obvious increased compared to model group(p＜0.01).Percentage of quadrant and 20%area of QYD treating group have no significant difference with Huperzine A-Zhulin Antun treating group,40%area has decreased compared to Huperzine A-Zhulin Antun treating group.3.The competence of information reserve of model group was obvious decreased,its percentage of quadrant have obvious decreased compared to normal group and blank group(p＜0.01),but 20%and 40%area have obvious increased compared to normal group and blank group(p＜0.01).Percentage of quadrant have obvious increased compared to model group(p＜0.01).Percentage of quadrant and 20%area have no significant difference with Huperzine A-Zhulin Antun treating group,40%area has decreased compared to Huperzine A-Zhulin Antun treating group.4.The frequency in span of platform localzation of model group has obvious decreased compared to other groups(p＜0.01).The frequency in span of platform localzation of QYD treating group has obvious increased compared to model group(p＜0.01).QYD treating group has increased compared to Huperzine A-Zhulin Antun treating group(p＜0.05).5.Apoptosis level of model group was obvious higher than other groups(p＜0.01).Apoptosis level of QYD treating group was obvious lower than Huperzine A-Zhulin Antun treating group(p＜0.01).6.Compared with normal group,the expression of bcl-2 and bax in model group decreased statistically(p＜0.01).Compared with model group,the expression of bax decreased in QYD treating group and Huperzine A-Zhulin Antun treating group(p＜0.01),the expression of bcl-2 increased(p＜0.01).Compared with Huperzine A-Zhulin Antun treating group,there were no significant differences of the expression of bax in QYD treating group,while the expression of bcl-2 increased(p＜0.01).Conclusion:1.Spleen kidney-Qi deficiency and phlegm stasis is the basic etiological factor for AD,and replenishing spleen kidney-Qi deficiency,removing phlegm therapy is operrative on treating AD.2.The Animal Model of Alzheimer's disease induced by composite Aβ_25-35and IBO injected into basal nucleus of Meynert of old rat simulated some characteristics of human AD to someextent,which maybe used as a novel model to study AD and its drug treatment.3.Qingqiao Yinao Decoction(QYD)could increase the expression of bcl-2 of hippocampus neural cells of rats,and decrease the expression of bax,and decrease apoptosis level to suppress the apoptosis of hippocampus neural cells of rats.4.Qingqiao Yinao Decoction(QYD)could improve disturbance learning and momory,while it also has obvious treating effection.