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  

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

作　　者：鲍华烨 周妙妮[1] 许爱娥[1] 

机构地区：[1]安徽医科大学附属杭州临床学院,杭州市第三人民医院皮肤科,杭州310009

出　　处：《中华皮肤科杂志》2014年第1期22-25,共4页Chinese Journal of Dermatology

基　　金：基金项目：国家自然科学基金（81071294,81271758）;浙江省自然科学基金（Z2100973）

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（rlL-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-13mRNA水平和上清蛋白水平。结果不同浓度（10、50、100、250、500txg／L）的rIL-13作用黑素细胞24、48、72、96h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-13mRNA（0．1000±0．0024）和蛋白[（1509．62±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：0．3832±0．0187，蛋白：（5507．98±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。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 lymphoeytes were isolated from both the tissue specimens and peripheral blood of these patients. DMSCs and melanoeytes 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 cocuhure 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 monoeultured CD8＋ T lymphocytes. C

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 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]