银屑病患者皮损间充质干细胞对HaCaT细胞增殖的影响

目的探讨银屑病患者皮损间充质干细胞（mesenchymal stem cells，MSCs）对HaCaT细胞增殖的影响，揭示银屑病可能的免疫发病机制。方法寻常性银屑病患者8例，进行期4例，静止期4例，银屑病皮损面积和严重程度（PASI）评分范围为1．6—18．0，平均11．64。健康对照组8例，取自我院泌尿外科和整形外科手术切除的正常皮肤。分离、培养患者皮损与健康人皮肤MSCs，流式细胞术进行细胞鉴定；将第5代皮肤MSCs与HaCaT细胞共培养，并设自然增殖组，采用实时细胞分析系统进行HaCaT细胞增殖检测，培养74h后用细胞计数法检测HaCaT细胞的数量。结果倒置显微镜下，银屑病组与健康对照组皮肤MSCs的细胞形态相似，细胞表面抗原均高表达CD29、CD44、CD73、CDg0、CDl05，而CD34、CD45及HLA—DR表达阴性。皮肤MSCs可抑制HaCaT细胞的增殖（P〈0．05），与健康对照相比，银屑病患者皮损MSCs抑制HaCaT细胞增殖的能力显著减弱[患者组细胞数为（2．35±0．254）×105，对照组细胞数为（2．04±0．122）×105，P〈0．05]。结论银屑病患者皮损MSCs抑制角质形成细胞增殖的能力减弱，这可能是银屑病的发病机制之一。

Effect of mesenchymal stem cells on HaCaT cell proliferation in patients with psoriasis lesions

Objective To explore the possible immune pathogenesis of psoriasis by investigating the effect of mesenchymal stem cells （MSCs） on HaCaT cell proliferation in patients with psoriasis lesions. Methods Eight healthy controls from Department of Urology and Plastic Surgery and 8 patients with psoriasis vulgaris（4 at progressive stage and 4 at resting stage） were recruited. The psoriasis area and severity index （PASI） score varied from 1.6 to 18.0 and averaged 11.64 in 8 patients. MSCs of passage 5 were obtained from psoriatic skin lesions and healthy human skins, and then identified using flow cytometry. The proliferation of HaCaT cell was investigated in a co-culture system by real-time cell analysis system and cell counting. The effect of MSCs on HaCaT cell proliferation was compared using one-way ANOVA and SNK test between patients and controls. The correlation of HaCaT cell quantity and PASI score was analyzed using Pearson correlation analysis. Results Inverted microscopy revealed that MSCs had similar morphology in two groups. Cell surface antigen expressed CD29, CIM4, CD73, CD90 and CD105 in patients with psoriasis vulgaris and controls, but not CD34, CIM5 or HLA-DR. Skin-derived MSCs inhibited HaCaT cell proliferation（ P 〈 0.05 ）. Compared with healthy control group, HaCaT cell proliferation increased in psoriasis group [（2.04＋0.122） ×105 vs （2.35＋0.254） ×105,P〈0.05]. Conclusion The results suggest that MSCs derived from psoriatic skin lesions have weaker inhibition effect on keratinocytes proliferation,which may be one of the pathogenic mechanisms of psoriasis.