中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (11): 779-783.

• 论著 • 上一篇    下一篇

银屑病患者皮损间充质干细胞对T细胞增殖的抑制能力减弱

刘瑞风,赵新程,杨元文,张开明   

  1. 太原市中心医院皮肤科
  • 收稿日期:2012-11-29 修回日期:2013-06-14 出版日期:2013-11-15 发布日期:2013-11-01
  • 通讯作者: 张开明 E-mail:zhangkaiming@sina.com
  • 基金资助:
    国家自然科学基金;国家自然科学基金

Diminished capacity of mesenchymal stem cells in lesions of patients with psoriasis to inhibit T cell proliferation

  • Received:2012-11-29 Revised:2013-06-14 Online:2013-11-15 Published:2013-11-01
  • Supported by:
    National Natural Science Foundation of China; National Natural Science Foundation of China

摘要: 【摘要】 目的 探讨银屑病患者皮损间充质干细胞(MSC)对T淋巴细胞增殖的抑制能力。 方法 寻常性银屑病患者15例,进行期7例,静止期8例。健康对照组15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MSC,流式细胞仪及多向分化法进行细胞鉴定;将第3代皮肤MSC与健康人外周血T细胞共培养,用细胞计数法及MTT法观察其对T细胞增殖的影响;ELISA法检测皮肤MSC培养上清液中白介素6(IL-6)、白介素11(IL-11)、肝细胞生长因子(HGF)和转化生长因子β1(TGF-β1)的浓度。患者组和对照组T淋巴细胞增殖比较采用单因素方差分析及SNK检验,细胞因子水平比较采用两独立样本均数比较的t检验。 结果 倒置显微镜下,银屑病组与健康对照组皮肤MSC的细胞形态和多向分化能力相似,细胞表面均表达CD29、CD44、CD73、CD90、CD105,而CD34、CD45及HLA-DR表达阴性。健康人外周血T细胞与银屑病患者及健康对照皮肤MSC共培养,T淋巴细胞计数健康对照组为(1.04 ± 0.29) × 105/ml,银屑病患者组为(1.67 ± 0.34) × 105/ml,两组比较,P < 0.01;MTT检测细胞增殖结果(A值)分别为0.275 ± 0.007和0.317 ± 0.021,两组比较,P < 0.01。培养上清液中,患者组MSC分泌IL-11水平(181.37 ± 31.74 ng/L高于健康对照组(130.07 ± 29.20 ng/L),两组比较,t = 5.32,P < 0.01;患者组MSC分泌IL-6(61.67 ± 17.53 ng/L)和HGF(319.24 ± 41.03 ng/L)水平低于健康对照组(分别为76.74 ± 18.96 ng/L和352.35 ± 51.47 ng/L),两组比较,t值分别为2.61和2.25,均P < 0.05);TGF-β1两组差异无统计学意义。结论 银屑病患者皮损MSC对T细胞增殖的抑制能力减弱,可能是银屑病的发病机制之一。 【关键词】 银屑病; 间质干细胞; T淋巴细胞

关键词: 银屑病, T淋巴细胞, 间充质干细胞

Abstract: LIU Rui-feng, ZHAO Xin-cheng, YANG Yuan-wen, ZHANG Kai-ming. Department of Dermatology, Taiyuan City Centre Hospital, Taiyuan 030009, China Correponding author: ZHANG Kai-ming, Email: zhangkaiming@sina.com 【Abstract】 Objective To evaluate the inhibitory effect of mesenchymal stem cells (MSCs) in lesions of patients with psoriasis on T lymphocyte proliferation. Methods Tissue specimens were obtained from the lesions of 15 patients with psoriasis vulgaris (7 at progressive stage and 8 at resting stage) and normal skin of 15 human controls from the Department of Urology and Plastic Surgery, Taiyuan City Centre Hospital. MSCs were isolated from these skin specimens, cultured, and identified using flow cytometry and in vitro differentiation assay. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the concentration of interleukin (IL)-6, IL-11, hepatocyte growth factor (HGF) and transforming growth factor (TGF)-β1 in the culture supernatant of third-passage MSCs. Peripheral blood T cells were obtained from a healthy adult and cocultured with the third-passage MSCs for four days. Then, cells were counted and methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferation of T cells. One-way analysis of variance (ANOVA) and Student-Newman-Keuls (SNK) test were carried out to compare the proliferation of T lymphocytes, and two independent samples t test to compare the concentrations of cytokines. Results Inverted microscopy revealed that the patient- and control-derived MSCs shared similar morphological properties and multi-directional differentiation capacity, along with the expression of CD29, CD44, CD73, CD90 and CD105, but absence of CD34, CD45 and HLA-DR on cell surface. After coculture with MSCs from the patients and controls for four days, the count of T lymphocytes per milliliter was (1.67 ± 0.34) × 105 and (1.04 ± 0.29) × 105 respectively (P < 0.01), and the proliferative activity (expressed as absorbence at 492 nm) was 0.317 ± 0.021 and 0.275 ± 0.007 respectively (P < 0.01). Compared with the control-derived MSCs, the patient-derived MSCs showed a significantly higher level of IL-11 ((181.37 ± 31.74) vs. (130.07 ± 29.20) ng/L, t = 5.32, P < 0.01), but a lower level of IL-6 ((61.67 ± 17.53) vs. (76.74 ± 18.96) ng/L, t = 2.61, P < 0.05) and HGF ((319.24 ± 41.03) vs. (352.35 ± 51.47) ng/L, t = 2.25, P < 0.05), as well as a similar level of TFG-β1, in the culture supernatant. Conclusions The inhibitory effect of MSCs in psoriatic lesions on T lymphocyte proliferation is diminished, which may contribute to the pathogenesis of psoriasis. 【Key words】 Psoriasis; Mesenchymal stem cells; T-lymphocytes