MicroRNA-155 Targeted Regulation Of IL32 Expression Promotes The Occurrence Of Atopic Dermatitis By Activating The JAK1/STAT Axis

Objective:.This study aims to explore the mechanism of micro RNA-155 affecting atopic dermatitis(AD)through interleukin-32(IL-32)/Janus-activated kinase-1(JAK1)axis.Methods: AD-related mRNA expression profile GSE124700(including 2 healthy controls and 2 skin tissue samples from patients with AD)and miRNA expression profile GSE62404(including 8 healthy controls and 8 blood samples from AD patients))were obtained through GEO database.GeneCards and STRING databases were used to screen genes related to AD.The GSE124700 analysis results and the top 100 disease genes in the GeneCards database were intersected,and the candidate genes were obtained using the Draw Venn Diagram tool.Go and KEGG enrichment analysis was performed for candidate genes by oebiotech biometric analysis tool.The upstream miRNAs of the target gene were predicted using the Targetscan and mirDIP databases,and the target relationship was verified by the dual luciferase experiment.Normal human serum samples(50 cases)and AD patient serum samples(90 cases)treated in our hospital’s dermatology department from June 2019 to May 2020 were selected for ELISA,and tissue samples from normal people and AD patients(10 cases)were collected to perform immunohistochemical staining.Human immortalized keratinocytes(HaCaT)were selected for in vitro cell experiments,qRT-PCR and Westren blot were used to detect the expression of pathway related factors.The expression of inflammatory factors was detected by qRT-PCR and ELISA.qRT-PCR,ELISA and H&E analysis was performed for the expression of inflammatory factors and histological changes in the in vitro AD skin remodeling model.Finally,a nude mouse model of AD was constructed to observe the occurrence of inflammation.Results: The differential analysis of GSE124700 and the results of GeneCards database found that interleukin family genes may play an important role in the occurrence and development of AD.GO and KEGG functional enrichment analysis found that the interaction of chemokine signaling pathways and cytokine receptors played an important role in the occurrence and development of AD.KEGG analysis further confirmed that IL32 may promote the occurrence and development of AD by activating the JAK-STAT signaling pathway.The prediction results of Targer Scan and mirDIP database found that miR-155 and miR-29 b may participate in the occurrence and development of AD through targeted regulation of IL32 expression.Dual luciferase experiments confirmed that miR-155 targeted and negatively regulated IL32.In AD,miR-155 was lowly expressed,and IL-32 was highly expressed in AD tissues and blood.IL-32 could up-regulate the expression of JAK1 and the phosphorylation level of downstream genes to activate the intracellular JAK pathway and promote the occurrence of AD.miR-155 could inhibit IL-32 to repress the activation of the JAK-STAT signaling pathway,and reduce inflammation in the AD skin remodeling model.In vivo experiments further confirmed that miR-155 inhibited the occurrence of AD by activating the IL-32/JAK-STAT axis.Conclusion: miR-155 may inhibit the activation of JAK-STAT signaling pathway by targeting the expression of IL32,thereby preventing the occurrence of AD.