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作 者:孙晓萱[1] 甘晓霞[1] 谈文峰[1] 魏禺[1] 华敏慧[1] 张缪佳[1]
机构地区:[1]南京医科大学第一附属医院风湿免疫科,南京210029
出 处:《现代免疫学》2013年第5期365-369,共5页Current Immunology
基 金:国家自然科学基金(30701129;30901332;81172845;81273294);江苏省自然科学基金(BK2011851;BK2012875)
摘 要:通过研究艾拉莫德抑制Th17细胞分化的调节作用,为临床治疗Th17相关性炎症与自身免疫病提供理论与实验依据。取C57BL/6小鼠脾脏淋巴细胞,磁珠分选CD4+CD62L+T细胞(nave T细胞),以anti-CD3、anti-CD28活化T细胞,同时加入IL-6、TGF-β及IL-23诱导nave T细胞向Th17细胞分化。在上述培养体系中加入艾拉莫德(53.4mmol/L)刺激细胞,并以空白组和MTX(0.3mmol/L)组作为对照。培养3d后收集细胞,流式细胞仪检测CD4+IL-17+Th17细胞的表达;酶联免疫吸附测定(Enzyme linked immunosorbent assay,ELISA)检测细胞培养上清白细胞介素-17(Interleukin-17,IL-17)水平;实时荧光定量PCR(real-time PCR)检测细胞IL-17和视黄酸相关的孤儿受体-γt(ROR-γt)基因表达水平。结果提示:在细胞水平,艾拉莫德对Th17细胞的抑制作用强于MTX;在蛋白水平,艾拉莫德对IL-17的抑制作用明显强于MTX(P<0.05);在基因水平,与MTX相比,艾拉莫德对ROR-γt mRNA和IL-17mRNA表达的抑制作用更强(P<0.05)。结果说明,艾拉莫德对Th17细胞的抑制作用强于MTX,艾拉莫德在控制RA病情中的作用可能优于MTX。To determine a role of Iguratimod in modulating Th17 cell differentiation, purified naive T cells(CD4+ CD62L+ T cells) from C57BL/6 mice spleen were cultured in wells precoated with anti-CD3 mAb and anti CD28 mAb, and subsequently stimulated with TGF-13, IL-6, IL-23 in order to induce differentiation of Th17 cells. The cells were divided into three groups, the experemental group that was treated with Inguratimod, the MTX control group and the normal control group. After 72 hours of culture, the frequencies of Th17 cells were measured by flow cytometry; the levels of IL-17 in the culture supernatants were measured by ELISA; the levels of IL-17 mRNA and ROR-γt mRNA were measured by real-time PCR. The results showed that the frequency of Th17 cells was significantly decreased in the experimental group compared with the MTX group and control group. IL- 17 level in the cultrue supernatant of the experimental group was significantly decreased as compared with that in the MTX group. ROR γt mRNA and IL- 17 mRNA were detected in both the experimental and MTX groups, and were higher in the experimental group than in MTX group. Taken together, we have identified a new function of Iguratimod, i. e. , decreasing Th17 cell differentiation. Suppression of IL-17 by Iguratimod may contribute to its potential role in RA therapy.
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