KXS Regulate Lipid Metabolic Disorders And Study Its Key Targets To Play Antidepressant Effect

Objective:To explore the effect of KXS on the metabolism of depression model in vivo and in vitro,to clarify the key targets and molecular mechanisms of KXS which plays an antidepressant role by regulating the metabolism,and to provide some experimental guidance for the research on the mechanism of antidepressant effect of KXS.Methods:1 chronic unpredictable mild stress rat models were synthesized,the rats in the model group,the Kaixinsan group,and the fluoxetine group were weighed and behaviorally evaluated before establishing models,4 weeks after establishing models and after 3 weeks of KXS treatment to analyze whether CUMS rats were successfully modeled and the effect of Kaixinsan on CUMS rats.Behaviorally tests include open field experiment,sugar water partiality test,forced swimming test.2 PT-PCR was used to detect mRNA expression levels 20 lipid-related genes(MMP2,MMP9,APOE,APOA1,APOB,APOD,APOC1,CLU,COL1A2,KDR,HSPB1,HSPA5,KNG1,VIM,PRG2 PLTP,PON1,SERPINA1,VCAM1,APP)in the hippocampus of CUMS rats before and after the intervention of KXS;Western Bolt technology was used to detect the expression levels of the above 20 genes in the hippocampus of CUMS rats before and after the intervention of KXS,and analyze effects of disorders of lipid metabolism of KXS on hippocampus of CUMS rats.3 establishments of corticosterone-damaged SH-Sy5 y cell models in vitro.By using the CCK-8 kit to detect the viability of SH-Sy5 y cells in the control group,model group and KXS group,detecting the effect of KXS on the viability of corticosterone-damaged SH-Sy5 y cells.4 PT-PCR technology was used to detect the mRNA expression levels of the above 20 genes in SH-Sy5 y cells after corticosterone-damaging before and after the intervention of KXS,further confirming the potential targets of KXS to regulate metabolic metabolism disorder and exert antidepressant effects at the cellular level.5 Using the Uniprot database website and DAVID data analysis software correlated the above 20 genes with known depression-related mechanisms.Using a string transfer database analyzed possible relationships between proteins and the signaling pathways that these proteins may participate in.Using Cytoscape database analysis software established a protein network to analyze the key targets of KXS to regulate lipid metabolism to exert antidepressant effects.6 The best sequence of SH-Sy5 y cells transfected with APOE siRNA was screened by PT-PCR technology,and PT-PCR technology was used to detect mRNA expression of 5 arthritis factors(IL-1?,IL-2,IL-6,TNF-?,COX-2)in SH-Sy5 y cells transfected with APOE siRNA before and after intervention of KXS,predicting a potential mechanism for KXS to regulate metabolic disorders and exert antidepressant effects.Results:1 After 4 weeks of establishments of CUMS rat models,the weight,sugar and water preferences,and total exercise distance of the control group rats were significantly higher than those of the other three groups,and the duration of forced swimming the other three groups were significantly lower than that of the control group.The reports have shown that CUMS rats have been successfully modeled.Three weeks after KXS administration,the weight and behavior scores of rats in KXS group and fluoxetine group reached normal levels,which proved that KXS has antidepressant effect.2 PT-PCR detected the mRNA expression levels of the above 20 genes in the hippocampus of CUMS rats before and after administration of KXS,and found that the expression of MMP2 and APOC1 mRNA was significantly reduced after KXS treatment,and MMP9,APOB,APOD,APOE,PON1,PLTP mRNA Expression increased significantly.Western Blot measured the protein expression levels of 7 genes with significant changes in mRNA,and found that after treatment with KXS,the expression of MMP2 protein was significantly reduced,APOE,PON1,PLTP expression was significantly increased.3 Using the CCK-8 kit to screen the modeling conditions and the concentration of KXS for corticosterone-damaged SH-Sy5 y cells,it was found that the cell viability was the lowest after 24 hours of 1 mmol/ L corticosterone-damaged SH-Sy5 y cells.After 500?g/ mL KXS intervention,SH-Sy5 y cell viability was restored.4 The mRNA expression levels of the above 20 genes of corticosterone-infected SH-Sy5 y cells were detected by PT-PCR before and after KXS administration.After treatment with KXS,MMP2,APOC1,HSPB1 mRNA expressions were significantly reduced,and APOB,APOD,APOE,HSPA5,PLTP,and VIM mRNA expressions were significantly increased.Synthesis of mRNA and protein expression of 20 genes in CUMS rats and corticosterone-injured SH-Sy5 y cells in vitro and in vivo suggests that MMP2,APOE,and PLTP may be potential targets for KXS to regulate lipid metabolism and exert antidepressant effects.5 Analysis of the protein network found that APOE not only has interaction with APOB,APOC,CALR,HSP90B1,APP,KNG1,CLU,SERPINA1,PON1,APOA,ABCA1 among 20 proteins,but also is related to the expression of IL-6 which is associated with the expression of inflammatory factors such as IL-2 and IL-1?.6 PT-PCR was used to analyse the mRNA expression of these inflammatory factors(IL-1?,IL-2,IL-6,TNF-?,COX-2)in SH-Sy5 y cells transfected with APOE siRNA1.The expression of IL-6,IL-2 and TNF-? mRNA changed significantly before and after the treatment of KXS,suggesting that the expression of APOE in SH-Sy5 y is directly related to the expression of IL-2,IL-6,TNF-?.The antidepressant effect may be caused by increasing the expression of APOE in the brain and then up-regulating the expression of the inflammatory factors IL-2,IL-6 and TNF-?.Conclusion: 1 KXS can exert antidepressant effects by regulating lipid metabolism disorders.MMP2,APOE,and PLTP may be potential targets for KXS to regulate lipid metabolism and exert antidepressant effects.2 The mRNA expression of APOE is directly related to the mRNA expression of IL-2,IL-6,and TNF-?.KXS may increase the mRNA expression of APOE in the brain,and then up-regulate the mRNA expression of inflammatory factors such as IL-2,IL-6 and TNF-? to exert antidepressant effect.