Effect Of Azelaic Acid On Psoriasis Progression Investigated Based On PI3K/AKT Signaling Pathway

Part One Multi-data biological information analysis of signaling pathways related to pathogenesis of psoriasisObjective:The aim is to uncover signal transduction pathways that are essential to psoriasis’s development through multi-data bioinformatics examination,and to supply theoretical backing for future research.Method:1.A comprehensive search of the gene expression database(GEO)(https://www.ncbi.nlm.nih.gov/gds/)for variations in gene expression analysis,linked to the psoriasis package,was conducted to construct a data set of differentially expressed genes(DEG)volcanic heat map figure and clustering analysis.2.A KEGG pathway enrichment map was plotted after a functional enrichment analysis of the common DEGs was conducted using the Kyoto Encyclopedia of Genes and Genomes.Results:1.Data sets GSE141804,GSE160932 and GSE164400 related to psoriasis were retrieved from GEO database.The GSE141804 dataset yielded a total of 215 DEGs,122 of which were up-regulated and 93 down-regulated.The GSE160932 dataset,meanwhile,had 135 DEGs,36 of which were up-regulated and 99 down-regulated.Lastly,the GSE164400 dataset had 292 DEGs,of which 195 were revised upwards and 97 downwards.2.An enrichment analysis of KEGG for common DEGs in GSE141804,GSE160932 and GSE164400 was conducted.GSEA analysis revealed that PI3K/mTOR signaling pathways were increased while autophagy signaling pathways were decreased in psoriasis.Conclusions:The GSEA analysis revealed a heightened expression of PI3K/mTOR signaling pathways and a decrease in autophagy signaling pathways in psoriasis,implying that the PI3K/AKT pathway is a significant factor in the development and progression of the disorder.Part Ⅱ In vivo study of azelaic acid on psoriasis progression and vascular proliferationObjective:To study the inhibitory effect of azelaic acid on psoriasis progression and vascular proliferation,and to investigate the possible mechanism.Methods:1.The back skin of mice was coated with imiquimod,the psoriasis model of mice was established and divided into groups.Azelaic acid was applied to the skin of the psoriasis model of mice,and the skin tissues were collected.2.The skin tissue structure of mice in each group was observed by employing Hematoxylin-eosin(HE)staining,and the effect of azelaic acid on psoriasis was evaluated.3.The skin tissues collected in groups were stained by immunohistochemistry,and the images were observed under a fluorescence microscope to analyze the fluorescence staining intensity of PI3K/AKT signaling pathway related proteins p-PI3K,p-mTOR and VEGF.4.Western Blotting revealed the expression levels of PI3K/AKT signaling pathway related proteins p-PI3K,p-AKT,p-mTOR,VEGF,COX-2,angiopoietin 1 and HIF-1α in the skin tissue of the mouse model,which had been extracted.Results:1.HE staining revealed the skin tissue structure of mice in both groups.Compared to the IMQ modeling group,azelaic acid treatment resulted in a thickening of dermal squamous epithelium,disordered cell arrangement,necrosis of collagen fibers,lymphocyte infiltration,and a decrease in dermal congestion.2.The psoriasis model group exhibited a notably heightened fluorescence intensity when immunofluorescence staining was employed to examine p-PI3K,p-mTOR and VEGF.In contrast,the azelaic acid treatment group’s fluorescence intensity was notably diminished in comparison to the psoriasis model group.3.The psoriasis model group exhibited significantly higher protein expression levels than the blank control group.In contrast,the azelonic acid treatment group exhibited significantly decreased relative protein expression levels of p-PI3K,p-AKT,p-mTOR,VEGF,COX-2,angiopoietin 1 and HIF1-α.Conclusions:1.Azelaic acid can significantly inhibit IMQ-induced local skin damage in mice with psoriasis.2.Azelaic acid may inhibit the activation of PI3K/AKT signaling pathway and improve the symptoms of psoriasis.3.PI3K/AKT signaling pathway is activated in IMQ-induced psoriasis,accompanied by the proliferation of angiogenic factors.Azelaic acid can inhibit the activation of PI3K/AKT signaling pathway and the proliferation of angiogenic factors,thus improving the skin lesions and vascular proliferation in psoriasis.Conclusions:1.Azelaic acid can significantly inhibit IMQ-induced local skin damage in mice with psoriasis.2.The activation of PI3K/AKT signaling pathway may be inhibited by azelaic acid,potentially leading to a decrease in the symptoms of psoriasis.3.The activation of PI3K/AKT signaling pathway,in IMQ-induced psoriasis,is accompanied by the proliferation of angiogenic factors.Azelaic acid,however,can inhibit this pathway and the proliferation of angiogenic factors,thus enhancing the skin lesions and vascular proliferation of psoriasis.Part Ⅲ In vitro the IL-17 model cells of HaCaT cells are inhibited in their proliferation by azelaic acidObjective:To construct reliable HaCaT cell lines to construct an IL-17 cell model,and to analyze the impact of azelaic acid on IL-17 cell proliferation and PI3K/AKT signaling pathway proteins.Methods:1.For CCK-8 cell proliferation/toxicity testing,stable experimental cells were cultivated,and cell viability was ascertained.Additionally,the survival rate of normal HaCaT cells and IL-17 model cells in varying azelaic acid concentrations was established.2.Western blotting revealed the expression levels of PI3K/AKT signaling pathway related proteins p-PI3K,p-AKT,p-mTOR,VEGF,COX-2,angiopoietin 1 and HIF-1α,which had been extracted from the self-cultured cells of each group.Results:1.No effect of azelaic acid on normal HaCaT cells was observed in the proliferation/toxicity test of CCK-8 cells,and varying concentrations of azelaic acid inhibited the proliferation of HaCaT cells significantly,with concentration gradient dependence.2.In the IL-17+azelaic acid group,a significant decrease in the relative protein expression levels of all related proteins was observed when compared to the blank group and negative control group,including p-PI3K,p-AKT,p-mTOR,VEGF,COX-2,angiopoietin 1,and HIF-1α.Conclusions:1.The follow-up experiment revealed that azelaic acid had a concentration of 5mmol/L and IL-17 at 100ng/ml,based on the experimental results and literature review.2.By inhibiting the PI3K/AKT signaling pathway and the proliferation of angiogenic factors,azelaic acid can ameliorate the signs of psoriasis.