Full-length Transcriptome Sequencing To Analyze The Effective Mechanism Of Differential Genes Of Oxymatrine In The Treatment Of Psoriasis

Objective In this study,full-length transcriptome sequencing was used to analyze the differential genes and pathways of oxymatrine before and after the treatment of psoriasis,and to explore the possible gene targets and pathways of oxymatrine in the treatment of psoriasis.Methods Sixteen consecutive patients with severe plaque psoriasis treated with oxymatrine for 8 weeks from July 1 to October 15,2019 were collected.5 patients were randomly selected for full-length transcriptome sequencing of skin lesions and normal skin tissues before and after treatment.The differential genes were analyzed by t-test,Wayne diagram was drawn for the results of differential analysis,and functional enrichment,pathway enrichment and protein interaction analysis were carried out.The core differential genes improved by oxymatrine treatment were verified by immunohistochemistry,Western blotting and real-time quantitative polymerase chain reaction.The psoriatic area and severity index（PASI）score and all genes were analyzed by weighted gene co-expression network analysis（WGCNA）and the co-expression module was constructed.The bright green module of top1in the five modules with the most significant PASI score was analyzed by functional annotation and protein interaction network analysis,and the core genes with the most significant improvement were identified by immunohistochemistry,Western blotting and real-time quantitative polymerase chain reaction.Results The number of DEGs in patients before and after oxymatrine treatment was4232,and 4105 DEGs were found between the psoriasis group vs.before oxymatrine treatment and the normal control group（P<0.01,|log₂fold change,（FC）|>1.5）.While most of the DEGs recovered signficantly after oxymatrine treatment,only 650 DEGs were observed between the psoriasis group（after oxymatrine treatment）and the normal control group（P<0.01,|log₂FC|>1.5）.Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway enrichment analysis showed that 64 pathways were significantly activated after oxymatrine treatment（P<0.05）.Only 12 pathways were statistically significant between after oxymatrine treatment and the normal control group（P<0.05）.Among all the restored pathways,the improvement of the IL-17 signaling pathway was the most significant（P=1.18E-06）.Gene loci of oxymatrine action was assessed by protein interaction analysis on 205DEGs that were co-expressed in 5 patients before and after oxymatrine treatment（P<0.05,FC>1.5）.After oxymatrine treatment,the expression of two mitosis-related genes namely,cyclin dependent kinase 1（CDK1）and cyclin B1（CCNB1）,that affect cell proliferation recovered significantly.The patients’PASI score and all genes were analyzed by WGCNA,and the co-expression module was constructed.Among the five modules with the most significant PASI score,the top1 bright green module was selected for functional annotation and protein interaction network analysis.The results showed that among a large number of genes related to oxymatrine regulation,the first five HUB genes were CNFN,S100A8,SPRR2A,SPRR2D and SPRR2E related to epidermal differentiation complex（EDC）.This suggests that oxymatrine may improve the differentiation of keratinocytes by regulating the expression of EDC-related genes.Oxymatrine can not only promote the development and repair of epithelial tissue,but also maintain the dynamic balance of keratinocytes.Conclusion Oxymatrine can change the abnormal expression of most genes and pathways in patients with psoriasis.Through multi-pathway and multi-target action,oxymatrine can significantly improve the abnormality of genes and pathways,so as to effectively treat psoriasis.Oxymatrine can inhibit the expression of proliferation-related genes CDK1 and CCNB1 and improve the abnormal proliferation in patients with psoriasis.Oxymatrine also regulates the epidermal differentiation complex（EDC）gene on 1q21 to maintain the homeostasis of cell differentiation.