银屑病患者皮损间充质干细胞生长缓慢的原因探讨

The research on the slow growth of dermal mesenchymal stem cells in patients with psoriasis

目的通过比较银屑病患者与健康人皮肤间充质干细胞(DMSCs)原代培养和3代总培养时间,及培养上清液中碱性成纤维细胞生长因子(bFGF)的浓度,研究DMSCs增殖速度与bFGF含量的关系。方法Dispase酶消化结合贴壁法分离培养患者与健康人皮肤DMSCs,流式细胞术及多向分化法进行细胞鉴定;细胞生长达90%融合时胰酶消化传代,记录原代培养时间与3代总培养时间,酶联免疫吸附试验(ELISA)法检测细胞培养上清液bFGF的浓度,bFGF对正常DMSCs增殖的影响。结果银屑病组与对照组皮肤DMSCs的细胞形态相似,均具有多向分化能力,细胞表面抗原CD105、CD44、CD73、CD29及CD90表达阳性,CD34、CD45及人类白细胞抗原(HLA-DR)表达阴性。与对照组相比,银屑病患者DMSCs原代培养和3代总培养时间延长(P<0.01),分泌的bFGF降低(P<0.01),加入bFGF组DMSCs原代培养和3代总培养时间缩短(P<0.01)。结论银屑病皮损DMSCs分泌bFGF水平下降,直接影响其增殖速度,这可能是银屑病患者间充质干细胞生长缓慢的原因。

Objective To explore the correlation between bFGF and the multiplication rate of dermal mesenchymal stem cells(DMSCs)derived from psoriatic lesions and normal controls psoriasis.Methods①To culture DMSCs by adhesion method;②To identify the DMSCs by flow cytometry and multiple differentiation method;③To detect bFGF in DMSCs cultural supernatants derived from psoriatic and normal controls by ELISA.Results The cell morphology and multiple differentiation ability of DMSCs from psoriatic group and normal control group were similar.The expression of cell surface antigen such as CD105,CD44,CD73,CD29 and CD90 was positive,while the expression of CD34,CD45 and HLA-DR was negative.Both the primary culture time and total culture time of psoriatic DMSCs were shortened compared with the normal controls(P<0.01).The bFGF level of psoriatic DMSCs were lower than that in the normal group(P<0.01).The bFGF can significantly improve proliferation rate of DMSCs.Conclusion DMSCs of psoriatic secreting lower bFGF might influence the proliferation ability of MSCs in patients with psoriasis.