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

作　　者：陈菲菲[1] 毛山[1] 史莹[1] 周丹阳[1] 张金梅[1] 谷伟[1] Chen Feifei;Mao Shan;Shi Ying;Zhou Danyang;Zhang Jinniei;Gu Wei(Department of Respiratory Medicine, Nanjing First Hospital, NMU, Nanjing 210006, China)

机构地区：[1]南京医科大学附属南京医院呼吸科,江苏南京210006

出　　处：《南京医科大学学报（自然科学版）》2019年第2期205-209,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京市卫生计委杰出青年基金(JQX16028)

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(30μg/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(1μmol/L)组、Zerumbone(10μmol/L)组、Zerumbone(30μmol/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(30μmol/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。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(30μg/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(1μmol/L)group;Zerumbone(10μmol/L)group;Zerumbone(30μmol/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(1μmol/L),Zerumbone(10μmol/L),Zerumbone(30μmol/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.

关 键 词：花姜酮 CD4^+CD62L^+T细胞 CD4^+CD25^+TREG 白细胞介素-10 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]