Study On The Mechanism Of Chai-Hu-Shu-Gan-San Regulating Aggressive Behavior In Mice

Objective: 1.Screening suitable methods for evaluating Chai-Hu-Shu-Gan-San(CSGS)to regulate the aggressive behavior of mice.2.The relationship between aggressive behavior and TLR4/NF-κB pathway was clarified,and the pharmacological mechanism of anti-aggression behavior exerted by CSGS evacuation through TLR4/NF-κB pathway was explored.3.The main metabolites and metabolic pathways of CSGS after intervention of aggressive behavior were preliminarily screened.Methods: 1.The aggressive behavior model of mice was established by residentintruder method,and the mice were divided into blank control group,model group,CSGS group and fluoxetine positive group.2.The changes of mouse attack behavior before and after the intervention of CSGS were evaluated by open field experiment,elevated cross maze experiment,and aggressive behavior test.3.Enzyme-linked immunosorbent and Western Blot(WB)were used to detect the content of related proteins in the mouse hippocampus in TLR4/NF-κB pathway.4.The level of IL-1β and activation of microglia in DG region of hippocampus were observed by immunofluorescence.5.Metabolomics was used to investigate the main metabolites and metabolic pathways in the hippocampus of mice before and after the intervention of CSGS.Results: 1.A mouse model of aggressive behavior was successfully established by the resident-intruder method.2.Behavioral results showed that the total distance of open field and the total distance of central area in model group increased significantly after drug administration.The time of opening the arm and the percentage of entering the open arm and the closed arm were significantly higher than those in the blank control group.In the model group,the incubation period of aggressive behavior was significantly shortened and the score of aggressive behavior was significantly increased.3.ELISA results showed that the content of IL-1β in the hippocampus of mice in the model group increased significantly,and the content of IL-1β decreased significantly after CSGS administration,close to the content of the blank group.The results of WB showed that compared with the blank control group,the expression of IL-1β protein in mice in the model group was significantly increased,and the expression content of TLR4,TNF-α,IL-1β and other proteins in the TLR4/NF-κB pathway was significantly reduced after CSGS administration.4.Immunofluorescence results showed that compared with the control group,the expression of IL-1β in the hippocampal DG region of mice in the model group increased,and the positive area of microglia increased significantly and activated.After CSGS treatment,the fluorescence intensity showed that the expression of IL-1β in the DG region decreased significantly to normal levels,inhibiting microglial activation.5.Metabolomic results showed that the main metabolic differences of the hippocampus before and after CSGS evacuation intervention in mice were D-Alanine,Deoxyinosine,7-Methylinosine,etc.,and the main metabolic pathways were arginine and proline metabolism,alanine,aspartic acid and glutamate metabolism,and glutathione metabolism.Conclusion: CSGS significantly improved the aggressive behavior of mice induced by residential invasion.The pathogenesis of aggressive behavior model mice is related to the contents of TLR4,TNF-α,NF-κB and IL-1β in the TLR4/NF-κB pathway.CSGS evacuation can inhibit the TLR4/NF-κB pathway activated by aggressive behavior and reduce the content of the above inflammatory factors,thereby inhibiting the occurrence of aggressive behavior.The production of aggressive behavior is related to the activation of microglia,and the evacuation of the liver can reduce the content of inflammatory factors in the hippocampus and inhibit the activation of microglia.After the intervention of CSGS in aggressive mice,the levels of phosphocreatine,mentheanine,proline betaine,1,9-nonylthiol,D-alanine,deoxyinosine and other metabolites were significantly changed.The anti-aggressive effects of CSGS were mainly related to the metabolism of arginine and proline,alanine,aspartic acid and glutamic acid,and glutathione.