Discussion On The Regulation Of Autophagy By 5-HT_1A Receptor In The Formation Of Depression Caused By Liver-stagnation And Spleen-deficiency And The Interventional Effect Of The Xiaoyao Powder Based On The TREK-1 Channel

ObjectiveIn this study,the chronic unpredictable mild stress?CUMS?was used to replicate the model of depression with liver-stagnation and spleen-deficiency,combined with the transcriptional and express-ional testing results of the nucleic acid and protein.The physio-logical,tissue,cellular and molecular levels confirmed the corre-lation between depression caused by liver-stagnation and spleen-deficiency with 5-HT_1A receptor protein,two pore domain potassium channel?TREK-1?and autophagy.This study aims to clarify whether the Xiaoyao Powder can affect the 5-HT_1A receptor,inhibit the autophagy response and reduce the neuronal apoptosis through the TREK-1 channel,and to further the potential mechanism of the Xiao yao Powder in the treatment of depression with liver-stagnation and spleen-deficiency.Methods1.Theoretical research:This study conducted literature re-search to understand the etiological causes and pathogenesis of depression in Chinese medicine,the classification of syndrome types in Chinese medicine,the pathogenesis of western medicine,and the progress of depression prevention and treatment with the Xiaoyao Powder.All these provide a solid theoretical foundation for the experimental research of this paper.2.Experimental research:Forty SPF grade male SD rats were divided into the normal group,the model group,the Xiaoyao Powder group and the fluoxetine group.The model of depression with liver-stagnation and spleen-deficiency was replicated with CUMS model-ing method for 7weeks.The general physiological indexes,sucrose preference test and open field experiment were used to evaluate the success of the model and efficacy of the Xiaoyao Powder.The Real-time fluorescent quantitative PCR?RT-qPCR?was used to detect and analyze the interventional effect of the well-known Chinese formula Xiaoyao Powder on the genetic transcription of the TREK-1channel and autophagy marker protein Beclin-1 and LC3 in the hippocampus of rats.The regulation of the Xiaoyao Powder on the protein expression of 5-HT_1A receptor,Beclin-1 and LC3 was detected by Western blot.The improvement of the Xiaoyao Powder on the apoptosis of hippocampal neurons in rats was determined by TUNEL staining.Results1.After successfully replicating the model of liver-stag-nation and spleen-deficiency depression with the CUMS modeling method,the rats showed the untidy and brown hair,decreased appe-tite,slow weight gain,reduced solitary activity and thin sloppy stool.In the open field experiment,the total distance and the central residence time of the rats were decreased and their rest time prolonged.The sucrose preference test showed decreased sucrose consumption ratio.The evidences indicate that the CUMS method successfully replicates the model of depression with liver-stagnation and spleen-deficiency,while the Xiaoyao Powder and fluoxetine have different degrees of improvement on the above pathological manifestations.2.Rt-qPCR results showed that the TREK-1 channel,Beclin-1 and transcriptional levels of LC3 in the model group,the Xiaoyao Powder group and the fluoxetine group were quite different and signifi-cantly higher than those in the normal group?P<0.01?.Compared with the model group,the transcriptional levels of the TREK-1 cha-nnel,Beclin-1 and LC3 in the Xiaoyao Powder group and the fluoxetine group were lower?P<0.01;P<0.05?.3.Western-blot results showed that,compared with the normal group,the relative gray values of Beclin-1 and LC3 proteins in the model group,the Xiaoyao Powder group and the fluoxetine group were significantly increased,while the relative gray value of 5-HT_1A receptor protein was decreased?P<0.01;P<0.05?.Compared with the model group,Beclin-1 and LC3 proteins expression were decreased in the Xiaoyao Powder group and the fluoxetine group?P<0.01;P<0.05?,while the relative gray value of 5-HT_1A receptor protein increased significantly?P<0.01?.The correlation analysis showed that the TREK-1 channel was positively correlated with Beclin-1 and LC3?P<0.01? but negatively correlated with 5-HT_1A receptor?P<0.01?.4.TUNEL results showed that the percentage of apoptotic nerve cells increased in the model group,the Xiaoyao Powder group and the fluoxetine group compared with the normal group?P<0.01;P<0.05?.Significantly decreased ?P<0.01?,the percentage of apoptotic nerve cells in the Xiaoyao Powder group was better than the fluoxetine group compared with the model group.Conclusion1.The literature research showed that liver-stagnation and spleen-deficiency occupies a very important position in the incidence of depression.2.The occurrence of the liver-stagnation and spleen-defi-ciency depression may be related to the influenced autophagy of the 5-HT_1A receptor protein through the TREK-1 channel,which regulates the autophagy marker proteins Beclin-1 and LC3 and finally leads to the neuron apoptosis.3.The Xiaoyao Powder regulates the 5-HT_1A receptor protein.The 5-HT_1A receptor protein influences autophagy through the TREK-1 channel,regulates the autophagy marker proteins Beclin-1 and LC3,reduces the apoptosis of neurons,and thus achieves the curative effect on depression caused by liver-stagnation and spleen-deficiency.