Research On Molecular Mechanism Of Depressive Phenotype In Mice From Generation To Generation As Well As Treatment Of Yueju Pill

Objective:To make a 4-week depressed phenotype of mice by chronic stress(restrainting 6h/day + overnight illuminating/2 times a week)or chronic corticosterone(CORT)hypodermic injection,and observe the depressive behaviors and the expression diversification of AKT-mTOR signal pathway in the offspring mice,and in quest into the passage mechanism of depressive phenotype mice and the therapeutic effect and mechanism of Yueju Pill on depression-like behaviors in offspring mice.Method:1.30 Balb/c mice(male/female)were randomly divided into control group(Con,15)and chronic stressed model group(Mod,15).The Con group were bred normally,while the Mod group were given chronic stresses,(restrainting 6h/d+ overnight illuminating/2 times a week)4w;30 C57BL/6N mice,?,randomly divided into control group(Con,15)and CORT model group(Mod,15),the Con group were breed normally while the Mod group were chronically injected with the CORT suspension at a dose of 20 mg·kg-1(CORT was first dissolved in DMSO,then added with 0.9%NaCl solution to make up a 2 g·L-1 suspension,and then mixed steadily with the ultrasonic vibration).To check whether the model is made successfully,behavioral tests were performed after the 4-week chronic stresses/injection,and the behavioral tests were performed at 3w,6w and lOw to observe behavioral changes.When the offspring mice born 10w,the hippocampus of the mice was removed and the change of expression of mTOR molecule was detected by western blot.2.After the depressive model of F0 was founded successful(hereinafter referred as ?mod,? mod,and CORT mod),the mice of each depressive model mate with the Con,and after the offspring mice were born(Con-F1,Mod-F1),behavioral tests were given at 3w,6w,low,and the hippocampus was taken out after the last time of the behavioral test to detect the change of expression of AKT/mTOR by western blot.3.The Balb/c model mice obtained from the previous experiment were divided into the F1 generation of female depressive model(?modF1)and the male depressive model(?mod F1).The two different models of F1 mice mate within their own model.And then F2 mice were born from female depression model(?modF2,Mod-F2)and the male depressive model(? modF2,Mod-F2),and the F1 Con mice were mated with Con(Con-F2).Finally the behavioral tests were performed at 3w,6w,and 10w after F2 were born.Immediately after these tests,the hippocampus was taken out to detecte change of expression of AKT/mTOR through western blot.4.In the second part,after the ?mod F1 female mice gorwn-up,picked out 10 randomly for 4w Chinese medicine Yueju Pills(2g/kg)treatment(YJ-Fl).The ?Con-F1 were mated one-toone with ?Con-Fl,?Mod-Fl,and YJ-F1 in adulthood,then Con-F2,Mod-F2 and YJ-F2 were born.The behavioral tests were performed at 3w,6w,and lOw after they were born.Detecting the changes of expression of AKT/mTOR western blot after these tests.Consequents:1.Chronic stressed model of the female and male mice,and the chronic CORT injection model mice all represent a depression-like behavior at 3w,6w,lOw behavioral tests of their offspring mice were born.There was a significant decrease in sucrose preference,a significant increase in immobility times in FST or TST,and a significant increase in latency and a significant reduction in eating weight in NSF.WB showed that no significant change in total protein expression of mTOR,but the phosphorylated protein and its ratio with total protein was significantly reduced.2.The F1 mice which experenced early life stress all showed depression-like behaviors at 3,6,and 10 weeks tests since they were born.WB showed that there is no significant change in total protein expression of AKT/mTOR,but the phosphorylated protein expression and their ratio with total protein were all significantly reduced.3.The F2 mice which under early life stress all showed depression-like behaviors at 3,6,and 10 weeks tests after born.WB showed that the there was no significance in total of AKT/mTOR expression in hippocampus,while the phosphorylated protein or their ratio with total protein were significantly reduced.4.The sugar performance of the female mice of ?modFl regained to normal after 4-week administration by Yueju Pill.The results of behaviors tests of F2 mice which were born by YJ-F1 at 3,6 and low,Compared with Mod-F2 showed that all of the depressive behaviors recover to normal,and the expression of AKT/mTOR in the hippocampus also returned to normal.Conclusion:1.Chronic stesses(binding 6h/d + night light 2 times/w)for 4w and CORT hypodermic injection for 4w all induce depression-like behaviors in their offspring mice,and the expression of mTOR protein in the hippocampus reduced significantly,this phenomenon could preserve 10w after the offspr:ing mice were born at least.2.Early life stress could induce depression-like behavior in F1 mice and there is an obviously decrease in the expression of AKT-mTOR pathway in hippocampus.This phenomenon could preserve 10w at least.3.Early life stress could induce depression-like behavior in F2 mice and there is an obviously decrease in the expression of AKT-mTOR pathway in hippocampus.This phenomenon could preserve 10w at least.4.Yueju Pills treatment of F1 female mice could regain the depressive behaviors of F2 mice which countered early life stress,and could also recover the expression of AKT-mTOR pathway in hippocampus.