Study Of Activity Constituents Of Kai Xin San In Vivo Based On Alzheimer’s Disease Model Rats

AimBecause the drug absorbtion was effected by the condition of the body,this study is to reveal direct material in vivo having an effect on Alzheimer’s disease,we conducted a systematic analysis of constituents in blood of AD model rat dosed with Kai Xin San,providing experimental data for the studies of pharmacodynamics material basis of Kai Xin San on Alzheimer’s disease and new constituents for curing Alzheimer’s disease.MethodsThe rat model of Alzheimer’s disease was made with the use of AlCl3 and D-gal.It was evaluated to the model and the therapeutic effects of Kai Xin San on the model by using cognitive ability and histology.Based on the effectiveness of Kai Xin San,ultra performance liquid chromatography coupled with high definition mass spectrometry(UPLC-HDMS/MS)and multiple energy data acquisition mode was applied to detect the constituents in in blood of AD model rat dosed with Kai Xin San.According to the identification results of Kai Xin San in vitro in the previous experiment,the constituents and its metabolites in blood of AD model rat dosed with Kai Xin San was characterized by direct comparison method and multivariate data analysis approach.Results1.The establishment and evaluation of AD model ratsThe rat model of Alzheimer’s disease was made with the use of gavage of AlCl3 and intraperitoneal injection of D-gal in period of 105 day.Compared with control group rats,model group rats were significant difference in cognitive ability test(Morris watertight palace experiment).The histopathological detection shows that the number of nerve cell was decreased,the expression of Aβ1-40 was increased,suggesting the AD model was successfully made.2.Study of evaluation of Kai Xin San effective on ADWhen the animal model was made,the rats were oraled with Kai Xin San.The study found that compared with model groups rats,the learning time was significantly shorten(P<0.05 or P<0.01)while the number of passing platform was significantly increased(P<0.01)for model rats dosed with Kai Xin San.Meanwhile,pathological and immunohisto-chemical observation showed that the number of nerve cells was increased and the expression level of Aβ1-40 was decreased.The results show that Kai Xin San exerted a improved effect on some indexs of AD rat model.3.Study of the activity constituents of Kai Xin San in vivo based on Alzheimer’s Disease ratsBased on the serum pharmacochemistry of TCM,an injection run of the sample of dosed model rat serum was completed in 21min with the developed method of the total components analysis of KXS.13 compounds were identified deriving from blood of AD model rat dosed with KXS.That are Adenosine,L-Uridine,S ibiricose Al,Ethyl benzoylacetate,Polygalaxanthone Ⅲ,2-Hydroxybenzonic acid,Asarylaldehyde,Dibutyl oxalate,α-Asarone,β-Asarone,Gominsin A and Tumulosic acid.Moreover,the consitituents of Poria and Rhizoma acori tatarinowii haven’t been reported under the condition of disease.ConclusionIn this paper,the rat model of Alzheimer’s disease was made with the use of AlCl3 and D-gal and we confirmed the validation of the intervention effect of KXS to AD model by cognitive ability test and histopathological detection.Under the direction of Serum Pharmacochemistry of TCM and UPLC-HDMS/MS technology,13 compounds were identified deriving from blood of AD model rat dosed with KXS.This study provides experimental data for research of pharmacodynamics material basis of Kai Xin San on Alzheimer’s disease and new drugs for curing Alzheimer’s disease.