中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (9): 616-619.

• 论著 • 上一篇    下一篇

谷氨酸信号通路在人外周血淋巴细胞活化及白癜风免疫发病机制中的作用

杨丽颖1,骆铁波1,单路娟2,刘越坚2,高鹤丽1,宋智琦1   

  1. 1. 大连医科大学附属第一医院皮肤科
    2.
  • 收稿日期:2011-10-13 修回日期:2012-03-17 出版日期:2012-09-15 发布日期:2012-08-31
  • 通讯作者: 宋智琦 E-mail:szqdalian@163.com
  • 基金资助:

    Glu信号通路在黑素细胞伪足形成及黑素小体转运中分子机制的研究;经典

Roles of glutamate signaling pathway in the activation of human peripheral blood lymphocytes and pathogenesis of vitiligo

  • Received:2011-10-13 Revised:2012-03-17 Online:2012-09-15 Published:2012-08-31
  • Contact: Zhiqi Song E-mail:szqdalian@163.com
  • Supported by:

    Study on molecular mechanisms effects of glutamate study on signaling pathway in filopodia formation, melanosome transfer in melanocyte

PDF

41

赞

摘要：

目的 探讨谷氨酸信号通路在人外周血淋巴细胞活化及白癜风免疫发病机制中的作用。方法 流式细胞仪测定离子型谷氨酸受体NMDAR的非竞争性拮抗剂MK801对健康对照外周血淋巴细胞CD25表达及IFN-γ水平的影响；NMDAR的激动剂NMDA和拮抗剂MK801对淋巴细胞内活性氧簇水平的影响。实时定量PCR和流式细胞仪测定白癜风患者和健康对照外周血淋巴细胞中NMDAR亚型NMDAR1和NMDAR2A表达差异。免疫组化方法观察白癜风患者皮损和健康对照皮肤组织中NMDAR1和NMDAR2A的表达。结果 MK801组外周血淋巴细胞CD25表达（7.28 ± 0.18）%较阴性对照组（16.02 ± 0.42）%明显下降（P < 0.01）；MK801组淋巴细胞中CD25+ IFN-γ+ 比例（1.79 ± 0.09）%较阴性对照组（0.78 ± 0.06）%明显增高（P < 0.01）。NMDA组淋巴细胞内ROS水平（101.1 ± 3.50）明显高于阴性对照组（69.80 ± 2.08）（P < 0.01）。白癜风患者外周血淋巴细胞NMDAR1的表达（3.85 ± 2.17）较健康组（0.97 ± 0.55）明显增高（P < 0.05）。结论 谷氨酸信号通路对活化淋巴细胞的IFN-γ分泌及ROS水平的影响有可能是白癜风的免疫发病机制之一。

关键词: 白癜风

Abstract:

Objective To investigate the roles of glutamate signaling pathway in the activation of human peripheral blood lymphocytes（PBLs） and pathogenesis of vitiligo. Methods Peripheral blood lymphocytes （PBLs） isolated from 5 patients with generalized vitiligo and 5 healthy controls were cultured in vitro. Flow cytometry was performed to quantify the expression of CD25 and interferon-γ on PBLs derived from healthy controls and treated with MK801 （a non-competitive antagonist of N-methyl-D-aspartic acid receptor, NMDAR） at 100 μmol/L or phosphate buffered saline （PBS） for 48 hours, as well as the level of reactive oxygen species （ROS） in the control-derived PBLs treated with MK801 at 100 μmol/L, NMDA （an agonist of N-methyl-D-aspartic acid receptor） at 0.5 mmol/L or PBS for 48 hours. The protein and mRNA expressions of NMDAR1 and NMDAR2A were measured by flow cytometry and real-time PCR, respectively, in PBLs from the healthy controls and vitiligo patients. Immunohistochemistry was used to observe the expressions of NMDAR1 and NMDAR2A in tissue specimens from depigmented and postinflammatory hyperpigmentation lesions of the patients with vitiligo and from normal skin of the healthy controls. Results Compared with the PBS-treated PBLs from the healthy controls, the MK801-treated PBLs showed a downregulated expression of CD25 （7.28% ± 0.18% vs. 16.02% ± 0.42%, P < 0.01）, but an upregulated proportion of CD25+IFN-γ+ lymphocytes （1.79% ± 0.09% vs. 0.78% ± 0.06%, P < 0.01）, and the NMDA-treated PBLs displayed a higher ROS level （101.1 ± 3.50 vs. 69.80 ± 2.08, P < 0.01）. The protein expression of NMDAR1 in PBLs was significantly higher in vitiligo patients than in the healthy controls （3.85 ± 2.17 vs. 0.97 ± 0.55, P < 0.05）. Conclusion Glutamate signaling pathway may be involved in the immunopathogenesis of vitiligo via affecting the secretion of interferon-γ by, and ROS level in, activated lymphocytes.

Key words: vitiligo