中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (1): 22-25.

• 论著 • 上一篇    下一篇

人真皮间充质干细胞对白癜风患者皮损周围CD8+ T淋巴细胞表达和分泌白介素13的影响

鲍华烨1,周妙妮2,许爱娥3   

  1. 1. 安徽医科大学杭州市第三人民医院皮肤科
    2. 杭州市第三人民医院皮肤科
    3. 安徽医科大学附属杭州市第三人民医院皮肤科
  • 收稿日期:2013-02-07 修回日期:2013-09-24 发布日期:2014-01-01
  • 通讯作者: 许爱娥 E-mail:xuaiehz@msn.com
  • 基金资助:
    国家自然科学基金;国家自然科学基金;浙江省自然科学基金

Effect of human dermal mesenchymal stem cells on the expression and secretion of interleukin-13 by perilesional CD8+ T lymphocytes from patients with vitiligo

Hua YeBAO1,Miaoni Zhou2,   

  • Received:2013-02-07 Revised:2013-09-24 Published:2014-01-01
  • Supported by:
    National Natural Science Foundation of China; National Natural Science Foundation of China

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摘要： 【摘要】 目的 探讨人真皮间充质干细胞（DMSC）对白癜风患者皮损周围CD8+ T淋巴细胞分泌表达白介素（IL）13的影响。 方法 细胞增殖检测法（MTS）检测重组IL-13（rIL-13）对黑素细胞增殖的影响。逆转录聚合酶链反应（RT-PCR）和Western印迹法分别检测6例进展期寻常型白癜风患者皮损周围及外周血中CD8+ T淋巴细胞中IL-13基因和蛋白表达。实时荧光定量反转录聚合酶链反应（qRT-PCR）技术和酶联免疫吸附试验（ELISA）分别检测DMSC与白癜风患者皮损周围CD8+ T淋巴细胞共培养前后细胞IL-13 mRNA水平和上清蛋白水平。 结果 不同浓度（10、50、100、250、500 μg/L）的rIL-13作用黑素细胞24、48、72、96 h后黑素细胞的增殖与对照组比较均未见明显变化（均P > 0.05）。白癜风患者皮损周围及外周血中CD8+ T淋巴细胞均可表达IL-13，但皮损周围CD8+ T淋巴细胞表达IL-13更显著。将皮损周围CD8+ T淋巴细胞和DMSC共培养，CD8+ T淋巴细胞IL-13 mRNA（0.100 0 ± 0.002 4）和蛋白［（1 509.62 ± 48.44） ng/L］的表达水平均显著低于单独培养的CD8+ T淋巴细胞［mRNA：0.383 2 ± 0.018 7，蛋白：（5 507.98 ± 34.11） ng/L，均P < 0.05］。 结论 白癜风患者皮损周围CD8+ T淋巴细胞高表达IL-13，DMSC能够有效地抑制其表达IL-13，或许可作为白癜风的治疗靶点之一。

关键词: 白癜风, CD8阳性T淋巴细胞, 间质干细胞, 白细胞介素13

Abstract: Bao Huaye, Zhou Miaoni, Xu Aie. Hangzhou Clinical College Affiliated to Anhui Medical University, Department of Dermatology, Third People′s Hospital of Hangzhou, Hangzhou 310009, China Corresponding author: Xu Aie, Email: xuaiehz@msn.com 【Abstract】 Objective To evaluate the effect of human dermal mesenchymal stem cells （DMSCs） on the expression and secretion of interleukin （IL）-13 by perilesional CD8+ T lymphocytes from patients with vitiligo. Methods Tissue specimens were obtained from the perilesional region of six patients with active vitiligo, and CD8+ T lymphocytes were isolated from both the tissue specimens and peripheral blood of these patients. DMSCs and melanocytes were obtained from the foreskin tissue of healthy males. The 3-（4,5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium, inner salt （MTS） assay was performed to estimate the effect of different concentrations of recombinant IL-13 on the proliferation of melanocytes, reverse transcripition-PCR and Western blotting to detect the mRNA and protein expressions of IL-13 in perilesional and peripheral blood CD8+ T lymphocytes respectively, real-time quantitative reverse transcription （RT）-PCR and enzyme-linked immunosorbent assay （ELISA） to detect the IL-13 mRNA expression in, and IL-13 protein expression in the culture supernatant of, CD8+ T lymphocytes before and after coculture with DMSCs, respectively. Statistical analysis was done by t test. Results No obvious changes were observed in the proliferation of melanocytes treated with different concentrations （10, 50, 100, 250, 500 μg/L） of recombinant IL-13 for various durations （24, 48, 72 and 96 hours） compared with untreated melanocytes （all P > 0.05）. Both perilesional and peripheral blood CD8+ T lymphocytes expressed IL-13, and the expression was stronger in perilesional than in peripheral blood CD8+ T lymphocytes. A significant decrease was noted in IL-13 mRNA expression （0.100 0 ± 0.002 4 vs. 0.383 2 ± 0.018 7, P < 0.05） and protein level in the culture supernatant （（1 509.62 ± 48.44） ng/L vs. （5 507.98 ± 34.11） ng/L, P < 0.05） of CD8+ T lymphocytes cocultured with DMSCs compared with monocultured CD8+ T lymphocytes. Conclusions There is a strong expression of IL-13 by perilesional CD8+ T lymphocytes in patients with vitiligo, which may be inhibited by DMSCs and serve as a target for the treatment of vitiligo.

Key words: Vitiligo, CD8-positive T-lymphocytes, Mesenchymal stem cells, Interleukin-13

引用本文

鲍华烨 周妙妮 许爱娥. 人真皮间充质干细胞对白癜风患者皮损周围CD8+ T淋巴细胞表达和分泌白介素13的影响[J]. 中华皮肤科杂志, 2014,47(1):22-25. doi:

Hua YeBAO Miaoni Zhou. Effect of human dermal mesenchymal stem cells on the expression and secretion of interleukin-13 by perilesional CD8+ T lymphocytes from patients with vitiligo [J]. Chinese Journal of Dermatology, 2014, 47(1): 22-25.doi: