A Study On Skin-derived Mesenchymal Stem Cells Biological Characteristicsand Differently Expressed MRNA In Patients With Psoriasis

Objective:This study investigate the biological characteristics of skin-derived mesenchymal stem cells,and the gene expression profile of SMSCs from patients with psoriasis. To reveal the abnormality of skin-derived mesenchymal stem cells (SMSCs)in psoriatic lesions microenvironment, and provide a theoretical basis for SMSCs involved in the pathogenesis of psoriasis.Methods:SMSCs were isolated and expanded by enzyme digestion from psoriatic lesions and normal control. To collect3rd passage of SMSCs and the culture supernatant. The cell morphology was observed under the inverted phase contrast microscope. The cell immunophenotypes were analyzed by flow cytometry. The multilineage differentiation potential of SMSCs were identified by adipogenic, osteogenic and chondrogenic induction system. Accumulative approach for drawing cell growth and proliferation curve.The level of EGF、TGF-β1in medium supernatant were detected by ELISA kits. Conducted mRNA expression profile and gene ontology analyses using microarray.Results:①Cellular morphology test:SMSCs morphology heterogeneity was observed under the inverted phase contrast microscope in both psoriatic lesions and normal control groups.②Cell surface marker:The third passage cells were detected by flow cytometry. Cell surface markes were expressed high for CD29、CD44、CD73、CD90and CD105,while were lower for CD34, CD45and HLA-DR.③Multilineage differentiation potential detection:The cells were induced with adipogenic, osteoplastic and chondrogenic differentiation medium for14days, and stained with Oil Red O, alizarin red S.and toluidine blue for staining adipocytes. osteocytes, and chondrocytes, respectively.④SMSCs growth and proliferation ability test:Cell growth and proliferation curve shows the third passage SMSCs proliferation rate in psoriasis group is faster than that of normal control.⑤Secreted cytokine levels (Epidermal growth factor and transforming growth factor-β1) detection:The level of EGF in patients was higher than that in control (P<0.05),and the level of TGF-(31in patients was lower than that in control (P<0.05).⑥Differently expressed genes analysis:Unsupervised hierarchical cluster analysis revealed that there was a difference in the expression patterns between the psoriasis and control groups.1927genes (1090up-regulated and837down-regulated) were found to show significantly different expression patterns.which the epigenetic regulatory factor UHRF1in psoriatic group was significantly enriched (P<0.05)Conclusions:①Stable method of isolation and culture of SMSCs from psoriatic lesions was established in our study, SMSCs morphology heterogeneity was observed as well.②bnormal Cellular biological characteristics existed in psoriatic SMSCs.③UHRF1in psoriatic group was significantly enriched by gene ontology analysis. Up-regulated UHRF1might be closely related with the biological characteristics abnormal changes in psoriatic SMSCs. SMSCs Abnormal epigenetic regulation involved in the pathogenesis of psoriasis.