文章摘要
陈菲菲,毛 山,史 莹,周丹阳,张金梅,谷 伟.花姜酮对小鼠CD4+CD25+Treg细胞分化功能的影响及机制[J].南京医科大学学报,2019,(2):205~209
花姜酮对小鼠CD4+CD25+Treg细胞分化功能的影响及机制
The effects of Zerumbone on the differentiation and functions of CD4+CD25+Treg cells
投稿时间:2017-09-17  
DOI:10.7655/NYDXBNS20190209
中文关键词: 花姜酮  CD4+CD62L+T细胞  CD4+CD25+Treg  白细胞介素⁃10
英文关键词: Zerumbone  CD4+CD62L+T cells  CD4+CD25+Treg cells  IL⁃10
基金项目:南京市卫生计委杰出青年基金(JQX16028)
作者单位
陈菲菲 南京医科大学附属南京医院呼吸科江苏 南京 210006 
毛 山 南京医科大学附属南京医院呼吸科江苏 南京 210006 
史 莹 南京医科大学附属南京医院呼吸科江苏 南京 210006 
周丹阳 南京医科大学附属南京医院呼吸科江苏 南京 210006 
张金梅 南京医科大学附属南京医院呼吸科江苏 南京 210006 
谷 伟 南京医科大学附属南京医院呼吸科江苏 南京 210006 
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中文摘要:
      目的:建立小鼠CD4+CD25+Treg细胞的体外诱导及培养方法,同时探讨花姜酮(Zerumbone)对Treg细胞的分化、分泌功能的影响及其机制。方法:从BABL/c小鼠的脾脏分离、纯化CD4+CD62L+T细胞,加入转化生长因子(transforming growth factor,TGF)? β(5 ng/mL)、 白细胞介素(interleukin,IL)?2(30 μg/mL)促进CD4+CD62L+T细胞向Treg细胞分化。Treg细胞被分为5组:正常组、模型对照组、Zerumbone(1 μmol/L)组、Zerumbone(10 μmol/L)组、Zerumbone(30 μmol/L)组。流式细胞术检测各组Treg细胞的比例,酶联免疫吸附试验(enzyme?linked immunosorbent assay,ELISA)方法检测IL?10含量,Foxp3、IL?10 mRNA的表达用实时荧光定量聚合酶链反应(RT?PCR)方法检测。结果:本实验方法使Treg细胞的诱导比例明显升高(P< 0.01)。与模型对照组相比,Zerumbone组的Treg细胞比例呈剂量依赖性升高(P均< 0.05)。IL?10蛋白的表达与模型对照组比较也呈剂量依赖性升高(P均< 0.05),Foxp3、IL?10 mRNA的表达同样升高(P < 0.05),其中Zerumbone(30 μmol/L)组升高最明显(P < 0.01)。结论:在体外实验中,Zerumbone可以促进BABL/c小鼠体内的CD4+CD62L+T细胞向Treg细胞分化,并促进IL?10蛋白的表达,其机制可能与诱导CD4+CD25+Treg特异性转录因子Foxp3的表达有关。
英文摘要:
      Objective:To explore an induced and culture method for CD4+CD25+Treg cells in vitro,study the effect of Zerumbone about the differentiation and secretion functions of Treg cells and explore the mechanisms included. Methods:CD4+CD62L+T cells were isolated from BALB/c mice spleen and purified with magnetic bead methods. CD4+CD62L+T cells were co?cultured with transforming growth factor(TGF)?beta(5 ng/mL),interleukin(IL)?2(30 μg/mL) for CD4+CD25+Treg polarization.The cultured CD4+CD25+Treg cells were divided into five groups:the normal group;the induced group,which were cultured with the above protocol;Zerumbone(1 μmol/L)group;Zerumbone(10 μmol/L)group; Zerumbone(30 μmol/L)group. Flow cytometry was used to detect the proportion of CD4+CD25+Treg cells. The ELISA method was detected the levels of IL?10. Reverse transcriptase polymerase chain reaction(RT?PCR)was detected the level of IL?10 mRNA and Foxp3 mRNA. Results:The proportion of CD4+CD25+Treg cells cultured with the protocol were significantly higher compared with the normal group(P < 0.05). The CD4+CD25+Treg cells proportion in Zerumbone(1 μmol/L),Zerumbone(10 μmol/L),Zerumbone(30 μmol/L)groups were significantly increased compared with group model,there is dose dependent(P < 0.05). The protein level of IL?10 was increased by Zerumbone and that was also dose?dependent. Zerumbone increased the expression of IL?10 mRNA and Foxp3 mRNA(P < 0.05). Conclusion:Zerumbone can increase the differentiation of splenic CD4+CD62L Zerumbone into CD4+CD25+Treg cells and induce the expressions of IL?10 protein in vitro. The results may be thought the activation of Foxp3 in CD4+CD25+Treg cells.
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