寻常型银屑病患者AMSCs的分离培养和免疫调控功能鉴定

目的对比寻常型银屑病患者和健康人来源的脂肪间充质干细胞(AMSCs)的表型、周期、成脂和成骨分化能力及免疫调控功能差异,探索患者AMSCs在银屑病发病机制中的作用。方法体外分离培养AMSCs,用流式细胞计量术检测AMSCs的细胞表型和周期;用油红O和碱性磷酶染色鉴定AMSCs的成脂和成骨分化能力;用PCR和ELISA检测AMSCs表达抑炎或促炎因子;通过基因表达谱芯片筛选两组AMSCs的免疫差异表达基因。结果健康人和银屑病患者AMSCs均高表达CD29、CD44及CD73和低表达CD45、CD31及HLADR。而且,都可分化成骨细胞和成脂细胞,但银屑病组AMSCs增殖趋缓(P0.05),成脂分化能力较强。银屑病组较对照组AMSCs表达或分泌的抑炎因子IL-10、IDO及TGF-β等下降(P0.05),而TNF-α及IFN-γ等促炎细胞因子增强(P0.05)。银屑病组AMSCs的JAK-STAT通路与对照组比明显下调(P0.05)。结论寻常型银屑病患者AMSCs较正常人的增殖和成脂分化能力出现异常,免疫抑炎调控能力减弱,推测可能与JAK-STAT免疫相关通路下调有关。

Isolating culture of adipose mesenchymal stem cells in psoriasis vulgaris patients and differentiation into immune regulation function

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.