| Purpose:Based on the previous achievements of the research group and the modern research on the pathological mechanism of AD,this study establishes the spleen qi deficiency syndrome model rats with cognitive impairment,and evaluates the model from the perspectives of fuzzy mathematics,behavior and pathology.Chinese medicine and western medicine treatment group are set up respectively for treatment of model rats,by testing organization level of inflammatory factors in the rats before and after medication,the lipid peroxidation product content and oxidase activity and TLR4/My D88/NF-κB signaling pathway target protein expression level,and investigate the effect of invigorating qi and strengthening spleen on improving neuroinflammation and peroxidation damage in cognitive-impaired rats from the perspective of dysfunction of spleen in transportation and the marrow sea deficiency,providing a theoretical basis for the principle of invigorating qi and strengthening spleen in the treatment of cognitive impairment.Material and method:A total of 60 SPF grade healthy SD rats(30 males and 30females)aged from 2 to 3 months were selected and randomly divided into the normal control group,the cognitive impairment model group of spleen qi deficiency syndrome,the positive drug control group and the group of invigorating qi,spleen and wisdom prescription.The model rats of spleen qi deficiency syndrome was duplicated by the combination of dietetic disturbance and excessive fatigue,and on this basis,Aβ1-42 hippocampal injection was carried out to establish the cognitive impairment model of spleen qi deficiency syndrome,and the model was evaluated by means of fuzzy mathematics,water maze behavior test and pathological observation.After continuous lavage treatment with invigorating qi,spleen and wisdom prescription and Aricept for 4 weeks,the whole brain and hippocampal tissues of the rats were taken.The levels of TNF-,il-6,MDA,SOD and other inflammatory and oxidative stress indicators in the brain tissues of rats were detected with the corresponding kits,and the protein expression and mRNA expression levels of TLR4,My D88 and NF-κB in the brain tissues of rats were detected by Western Blot and real-time quantitative PCR respectively.Results:1 The changes in body weight and food intake of rats: During modelling,compared with the normal group,the growth rates of body weight and food intake of rats in the model group(i.e.,the model group,the western medicine group,the TCM group,the same below)were all reduced.During treatment,compared with the model group,the growth rates of the amount of food and weight of rats in the western medicine group and the TCM group were all increased.2.Behavioral detection results of rats: The training results of positioning navigation showed that the escape incubation period of the model group was increasing compared with that of the normal group.The experimental results of space exploration showed that the proportion of platform quadrant activity time and times of crossing platform area of the modeling group were significantly less than those of the normal group(P<0.01),and the time to reach the platform area for the first time was significantly longer than that of the normal group(P<0.01).3.Histopathological examination of rat brain: After HE staining in the hippocampal area of the rat brain,neurons in the normal group were arranged in a regular and large number,and the staining was symmetrical.The nucleus is round and the nucleoli are obvious.In the model group,the number of pyramidal cells decreased,the arrangement was disordered,the gap was widened,the nuclei were hyperchromatic,the cytoplasm was wrinkled,a large number of glial cells were proliferated,and the surrounding tissues formed edema.Compared with the model group,the tissue morphology of the western medicine group and the TCM group recovered significantly after drug administration,the number of cells increased,the arrangement was more regular,the nuclear staining was relatively clear,and the degree of glial cell proliferation was also significantly improved.4.The detection results of inflammation and oxidative stress indicators in the brain tissues of rats in the model group showed that the contents of TNF-α、IL-6、MDA in the brain tissues of rats in the model group were significantly increased compared with those of the normal group(P<0.01),and SOD activity was significantly decreased(P<0.01).5.Results of TLR4,My D88 and NF-κB protein expression in brain tissues of rats: Western Blot results showed that the protein expression levels of TLR4,My D88 and NF-κB weresignificantly increased in model group compared with normal group(P<0.01).Compared with the model group,the protein expression levels of TLR4,My D88 and NF-κB in the western medicine group and the TCM group all decreased to different degrees(P<0.01).6.Results of TLR4,My D88,NF-κB mRNA expression in rat brain tissues: the results of real-time fluorescence quantitative PCR showed that the mRNA expression levels of TLR4,My D88,NF-κB in the model group were significantly increased compared with the normal group(P<0.01).Compared with the model group,the mRNA expression levels of TLR4,My D88 and NF-κB in the western medicine group and the TCM group all decreased to different degrees(P<0.01).Conclusions:1.The structure,number and arrangement of brain cells in rats with the spleen qi deficiency syndrome with cognitive impairment were changed,which constituted the pathological basis for the occurrence of cognitive impairment.2.The neuroinflammatory response and oxidative stress response in the brain of cognitive impairment rats were continuously activated,leading to the production of a large number of inflammatory factors and oxygen free radicals,which promoted the degeneration of nerve cells and the formation and deposition of amyloid protein.This process is involved in the modern medical pathogenesis of cognitive impairment.3.As one of the pathogenesis of cognitive impairment in traditional Chinese medicine,spleen deficiency promote the inflammatory reaction and lipid peroxidation damage in the brain of cognitive impairment rats.The therapy of invigorating qi and strengthening spleen can regulate the activity degree of TLR4/My D88/NF-κB signaling pathway,reduce the inflammation and peroxidation damage in the brain of rats with cognitive impairment,and delay the pathological process of cognitive impairment. |
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