Effect Of Guben-jiannao Method In Rat With AD Based On The Vesicular Transport Pathway Of APP

Objective To explore the theoretical basis for the prevention and treatment of AD by the method of Guben-Jiannao method based on the theory of Spleen and stomach are the acquired foundation,and to establish a AD model to observe the effect of Guben-Jiannao method on AD,providing a theoretical basis and experimental basis for the prevention and treatment of AD by traditional Chinese medicine.Methods1.theoretical research: The literature study method was used to systematically review the understanding of traditional Chinese medicine on Alzheimer’s disease.Based on the theory of "the congenital and acquired foundation",we expound Guben-Jiannao method about therapeutic principle and therapeutic method with our related research results.2.experimental study:90 SPF elderly(12 month old)male Wistar rats were randomly divided into 7 groups: normal group,sham operation group,model group,high dose group of GBJN,median dose group of GBJN,low dose of GBJN and positive control group after one week adaptation with general meals.The normal group were not modelled.The other 5 groups were handled with cerebral stereotactic surgery that was injected into the bilateral hippocampal CA1 area to make the compound AD rat model by Aβ1-42.The sham operation group was injected with the equal volume of normal saline.After 3 days,the treatment groups were treated with the GBJN decoction,positive control group with Donepezil Hydrochloride Tablets,sham operation group and blank control group with equal volume of normal saline.After 4 weeks,Morris water maze were used to test the spatial learning and memory ability of rats.Western blot method was used to detect the expression of Sor LA and VPS35 in each group.The expression quantity and correlation of Sor LA,VPS35 and APP were detected by Immunohistochemistry.And the expression of Aβ1-42 was detected by ELISA method,so as to explore the effect and mechanism of Guben-Jiannao method in preventing and treating AD.Result1.The rat model of AD rats with impaired learning and memory ability can be induced by injecting Aβ1-42 into the bilateral hippocampus of rats.2.The Morris water maze test showed that the median dose of GBJN decoction could improve the learning and memory ability of rats.There was significant difference on learning and memory ability,the model group was decreased compared with the sham operation group(P<0.01).After treatment,the learning and memory ability of rats has been improved.Compared with the model group,the escape latency on each treatment group was significantly shortened,and the number of crossing platform was increased,there was significant difference on each group(P <0.05).Compared with the postive control group,the escape latency of the median dose group was shortened and the number of crossing platform was increased,the curative effect was better than that in the postive control group,there was significant difference on each group(P < 0.05).3.Western blot was used to detect the expression of SorLA and VPS35..Compared with the sham operation group,the expression of VPS35 and Sor LA in the model group was decreased significantly(P < 0.01).After treatment,the expression of VPS35 and Sor LA in each treatment group was improved compared with the model group,(P<0.01).Compared with the postive control group,the expression of VPS35 in the median dose group was increased,and the expression of Sor LA in the high dose group was improved(P < 0.05).4.The IF single marker results showed that the SorLA red fluorescent particles and the VPS35 green fluorescent particles in the model group were less than the sham group(P < 0.01).Compared with the model group,more red and green fluorescent particles were seen in each treatment group(P < 0.05).The IF double marker results showed that compared with sham operation group,decoctionllow fluorescent particles in model group were fewer than those in sham operation group,and decoctionllow fluorescent particles were made up of overlapping red Sor LA fluorescent particles and green VPS35 particles(P<0.01).Compared with the model group,there were more decoctionllow fluorescent particles in each treatment group(P < 0.05).5.The expression of Aβ1-42 in the hippocampus of rats before and after the detection of ELISA.After the drug intervention,the expression of Aβ1-42 in each group was reduced,and the expression of Aβ1-42 in the GBJN decoction was the lowest.Conclusion1.The connotation of the Guben-Jiannao method aims at consolidating the congenital and acquired foundation,.nourishing the brain and spirit,eliminating phlegm and removing stasis.The mechanism of ad caused by spleen and stomach weakness may be:once the spleen and stomach were weak,reducing deficiency of five zang organs,obstruction of collaterals with phlegm and stasis,losing nourishment in brain.Invigorating the spleen and kidney,eliminating phlegm and opening the orifices are the therapeutic methods.2.Guben-Jiannao method can improve the learning and memory ability of dementia rats and improve the cognitive function of dementia rats.3.One of the mechanisms of Guben-Jiannao method on prevention and treatment of dementiais may be preventing Sor LA and VPS35 in the process of vesicular transport,which will lead to more APP entering into the non amyloidosis pathway and decrease the production of Aβ.