细粒棘球蚴囊液对体外培养小鼠脾细胞Foxp3及Smad4基因表达的影响  

The influence of Echinococcus granulosus fluid on the expression of Foxp3 and Smad4 genes of mouse spleen cells in vitro

作　　者：周洋[1,2] 金一帮[1,2] 王俊华[1] 吐尔洪江·吐逊[1,2] 魏晓丽[1] 张志[1,2] 张传山[1] 李亮[1] 林仁勇[1] 温浩[1,2] 

机构地区：[1]新疆医科大学第一附属医院医学研究中心,新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院消化血管外科中心

出　　处：《中国病原生物学杂志》2011年第3期193-196,182,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.30960342);新疆维吾尔自治区自然科学基金青年基金项目(No.2010211B19);新疆包虫病基础医学重点实验室开放课题项目(No.XJDX0202-2009-03)

摘　　要：目的观察细粒棘球蚴囊液对体外培养BABL/c小鼠脾脏细胞中调节性T细胞(Treg细胞)相对特异分子Foxp3(叉头蛋白3)及TGF-β1(转化生长因子-β1)下游信号通路Smad4表达的影响。方法以BABL/c小鼠作为脾细胞供者,用研磨法分离脾脏细胞,随机分为实验组(与细粒棘球蚴囊液共同培养)和对照组,取1×105个细胞接种于96孔板上,分别在1、3、6和12 h提取RNA,实时荧光定量PCR(qRT-PCR)检测Foxp3以及Smad4的基因表达。结果 qRT-PCR显示,细粒棘球蚴囊液处理1、3和12 h,实验组Foxp3表达量分别为4.577±0.317、8.517±0.978和7.406±0.822,对照组分别为9.274±0.451、3.297±0.408和2.464±0.328,差异均有统计学意义(P<0.05);细粒棘球蚴囊液处理1 h和12 h,实验组Smad4表达量分别为3.862±1.417和1.690±0.248,对照组分别为1.689±0.221和3.600±1.081,差异有统计学意义(P<0.05),且Foxp3与Smad4二者之间存在负相关(r=-0.991,P<0.05)。结论细粒棘球蚴囊液对小鼠脾脏细胞中Treg相对特异分子Foxp3有上调作用,而对TGF-β1的下游信号通路Smad4有下调的作用,二者之间存在负相关,提示细粒棘球蚴侵染宿主的过程中可能通过负调控TGF-β信号通路而造成宿主Treg细胞的表达升高,可能参与细粒棘球蚴感染过程中的免疫逃避。Objective To observe the expression of Foxp3 and Smad4 in the spleens of BABL/c mice treated with Echinococcus granulosus fluid（EgF）.Methods Spleen cells of BABL/c mice were isolated by grinding and randomly divided into an experimental group（treated with EgF） and a control group.Total RNA was extracted at 1,3,6,and 12 h,and the gene expression of Foxp3 and Smad4 was detected by real-time quantitative PCR（qRT-PCR）.Results qRT-PCR results showed that the expression of Foxp3 at 1 h,3 h,and 12 h was 4.577±0.317,8.517±0.978,and 7.406±0.822 for the experimental group while expression of Foxp3 was 9.274±0.451,3.297±0.408,and 2.464±0.328（P0.05） for the control group.The expression of Smad4 at 1 h and 12 h was 3.862±1.417 and 1.690±0.248 for the experimental group while expression of Smad4 was 1.689±0.221 and 3.600±1.081（P0.05） for the control group.Results indicated a negative correlation between Foxp3 and Smad4（r=-0.991,P0.05）.Conclusion The expression of Foxp3 increased significantly with decreased expression of Smad4 after treatment with EgF,suggesting that Treg cells may be down-regulated by the TGF-β signaling pathway and E.granulosus may be engaged in immune evasion.

关 键 词：细粒棘球蚴 囊液 脾细胞 FOXP3 SMAD4 

分 类 号：R383.34[医药卫生—医学寄生虫学]