Effect Of Xiaoyao San Against Chronic Stress-induced Injury On FGFR1,PLA2G3 And PLA2G5 Gene Expression Of MAPK Signaling Pathway In Hippocampus Of Rats

Objective To study the effect of Xiaoyao San on the mRNA expression and translation levels of FGFR1,PLA2G3 and PLA2G5 genes in the extracellular signaling kinase(ERK/MAPK)pathway in the hippocampus of chronic stress depression model rats,so as to explore the mechanism of Xiaoyao San in improving chronic stress injury on ERK/MAPK signaling pathway.Method 120 SPF grade male Wistar rats were randomly divided into blank control group,chronic stress model group,Xiaoyao San intervention group and western medicine prescription Kaike(fluoxetine)intervention group,with 30 rats in each group.The blank control group was not subjected to stress stimulation.The other three groups adopted chronic unpredictable stress(CUMS)rat model.Xiaoyao San intervention group and Kaike(fluoxetine)intervention group were respectively given 10ml/kg of Xiaoyao San and 10ml/kg of Kaike aqueous solution during the modeling process,and set three modeling times as 21 d,28d and35 d.After continuous modeling,rats were sacrificed on the 22 nd day,the 29 th day and the 36 th day,respectively,and all indexes were determined.The results of serum corticosterone were tested by Elisa,and the experimental animal behavior(weight test,open field test)was used to evaluate the success of chronic stress model.The mRNA and protein expression of FGFR1,PLA2G3 and PLA2G5 genes in the hippocampal area of each group were determined by fluorescence quantitative RT-PCR and western-blot assay.Results1.Behavioral results:The weight of the rats in each group was lower in the chronic stress group(B)than in the blank control group(A)and the Xiaoyao San intervention group(C)(P<0.05),while the 21-day experimental period in the Kaike intervention group(D)was significantly greater than that in the chronic stress group(B)(P<0.01).However,in the 28-day and 35-day experimental cycles,it was smaller than that in the blank control group(A),the chronic stress group(B),and the Xiaoyao San intervention group(C)(P<0.05).In the open box experiment,the number of crawling lattice and the number of standing position within 3minutes were smaller in the chronic stress group(B)than in the blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.05),and were also significantly smaller in the Xiaoyao San intervention group(C)and the Kaike intervention group(D)than in the blank control group(A)(P<0.05).2.Serum corticosterone test results:At 21 d,28d and 35 d,the serum corticosterone concentration in the chronic stress group(B)was greater than that in blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.01).The Xiaoyao San intervention group(C)and the Kaike intervention group(D)were larger than the blank control group(A)(P<0.01).The Xiaoyao San intervention group(C)was also larger than the Kaike intervention group(D)(P<0.05).3.The real-time fluorescent quantitative PCR detection results:The three time periods of21 d,28d,35 d between groups in the rat hippocampus gene transcription of mRNA,the FGFR1,PLA2G5 mRNA expression of the chronic stress group(B)were significantly less than the blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.01),the PLA2G3 mRNA expression of the chronic stress group(B)were greater than the blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.01).The mRNA expression levels of PLA2G5 and PLA2G3 genes at 21 d,28d and 35 d,the chronic stress group(B),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)were significantly different(P<0.01).4.Western blot results:The three time periods of 21 d,28d,35 d between groups in the rat hippocampus translation protein expression level,the FGFR1,PLA2G5 protein expression of the chronic stress group(B)were significantly less than the blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.05),the PLA2G3 protein expression of the chronic stress group(B)were greater than the blank control group(A),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)(P<0.05).The protein expression levels of PLA2G5 and PLA2G3 genes at 21 d,28d and 35 d,the chronic stress group(B),the Xiaoyao San intervention group(C)and the Kaike intervention group(D)were significantly different(P<0.01).Conclusion1.From general behavioral observation,body weight test,open box experiment,tail suspension experiment and serum corticosterone test results,it was verified that Xiaoyao San and Kaike(fluoxetine)could significantly improve chronic stress and depression in rats.According to the results of this study,it is preliminarily concluded that Xiaoyao San is better than Kaike(fluoxetine),has a long-lasting effect,is not easy to resist,and has higher safety.2.In three periods of 21 d,28d,35 d,chronic stress may significantly down-regulate the expression of FGFR1,PLA2G5 mRNA and its translation protein,the expression of PLA2G3 mRNA and the corresponding protein were detected in MAPK signaling pathway.It can be confirmed that chronic stress can cause depression by affecting the abnormal expression of MAPK signaling pathway.3.Xiaoyao San can improve the symptoms of anxiety,tension,depression,insomnia and other neurological and mental disorders caused by chronic stress stimulation through the intervention of MAPK signaling pathway.