Study On The Mechanism Of Acupuncture In The Treatment Of Qi Stagnation Constitution Based On Transcriptome Sequencing

Objective: This research work is based on transcriptome sequencing for data analysis.Objective 1: to screen the differentially expressed genes of qi stagnation constitution,and use bioinformatics methods to analyze the gene function and pathway enrichment of the differentially expressed genes;Objective 2: to use acupuncture to intervene the subjects of qi stagnation constitution,and analyze the transcriptome sequencing,compare the gene differential expression characteristics before and after acupuncture intervention,preliminarily reveal the function of gene differences related to qi stagnation constitution,and explore the effective mechanism of acupuncture intervention on qi stagnation constitution.Methods:1.From October 2020 to August 2021,31 cases of qi stagnation constitution and 30 cases of balanced constitution were recruited from the treatment and prevention center of the Affiliated Hospital of Shandong University of traditional Chinese medicine.According to the constitution type,they were divided into qi stagnation constitution group and balanced constitution group,and the fasting peripheral blood of the two groups were sequenced.The differentially expressed genes of qi stagnation constitution group were screened,and the function enrichment and signal pathway enrichment of differentially expressed genes were analyzed;2.Acupuncture intervention was performed on 31 subjects in qi stagnation constitution group.Acupuncture Baihui,Yintang,Shenmen,Taichong,Neiguan and Danzhong for 30 min each time,three times a week.After 8 weeks of treatment,the fasting peripheral blood was sequenced.The differentially expressed genes after acupuncture intervention were screened by comparative analysis with those before treatment,and the functional genes and action pathways of acupuncture intervention on qi stagnation constitution were screened by bioinformatics analysis.Results:1.Comparing qi stagnation constitution group with balanced constitution group,308 differentially expressed genes were screened,including 285 up-regulated genes and 23down-regulated genes.Bioinformatics analysis showed that the differentially expressed genes of qi stagnation constitution were significantly enriched in thyroid hormone synthesis and metabolism,glycosaminoglycan binding,cell adhesion molecules,extracellular matrix,chemokines,axon guidance,PI3 K Akt signaling pathway and TGF-β signal pathway,c AMP signal pathway,etc.Gene set enrichment analysis was supplemented to obtain serotonin transport,response to corticosterone,acetylcholine receptor activity,response to acetylcholine,acetylcholine gated channel complex,G protein coupled acetylcholine receptor signal pathway,etc.APLNR、SPARCL1、CLDN16、ADAMTS15、CNTN4and other seed node genes were obtained by protein-protein interaction(PPI)analysis.2.Compared before and after acupuncture intervention,qi stagnation constitution’s score,SDS score,HAMD score,HAMA score and PSQI score were significantly lower,and the change was statistically significant(P < 0.001).313 differentially expressed genes after acupuncture intervention were screened by transcriptome sequencing,including 61 upregulated genes and 252 down-regulated genes.Bioinformatics analysis showed that after acupuncture intervention,differentially expressed genes were significantly enriched in collagen containing extracellular matrix,thyroid hormone synthesis and metabolism,transmembrane receptor protein tyrosine kinase signal pathway,glycosaminoglycan binding,axon guidance,extracellular matrix receptor interaction,cell adhesion molecules,cytokinecytokine receptor interaction PI3 K Akt signaling pathway,TGF-β signaling pathways and cancer related pathways.COL1A2、CLDN16、APOE、APLNR seed node genes were obtained by intersection screening and PPI analysis,and after acupuncture intervention,the expression tended to be balanced constitution.Conclusions:1.There are significant differences between qi stagnation constitution and balanced constitution at the molecular level.Its biological process and signal pathway involve neuroendocrine,neuroplasticity,neurotransmitter and cholinergic system,which are mainly affected by TGF-β Regulation of signal,PI3 K Akt signal pathway and c AMP signal pathway.The key genes are APLNR、SPARCL1、CLDN16、ADAMTS15、CNTN4.2.Acupuncture intervention can effectively improve qi stagnation score,alleviate anxiety and depression,and improve sleep quality.Its mechanism is related to the improvement of neuroendocrine and neuroplasticity in patients with qi stagnation constitution,and may be through TGF-β signal pathway and PI3 K Akt signal pathway play a regulatory role.COL1A2、CLDN16、APOE、APLNR may be the functional genes of acupuncture intervention on qi stagnation constitution,and after acupuncture intervention,the expression tended to be balanced constitution.