IL-10表达对小鼠脾脏CD4+T细胞亚群相关细胞因子的影响  

The effect of IL-10 expression on CD4+T cell subset related cytokines in mouse spleen

作　　者：李东杰[1] 薛永平 刘丽媛 陆思敏 程锁利[2] LI Dongjie;XUE Yongping;LIU Liyuan;LU Simin;CHENG Suoli(Department of Clinical Laboratory,People′s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,China;Department of Joint Surgery,People′s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,China;The Third Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏回族自治区人民医院临床医学检验诊断中心,宁夏银川750002 [2]宁夏回族自治区人民医院关节外科,宁夏银川750002 [3]宁夏医科大学第三临床医学院,宁夏银川750004

出　　处：《宁夏医学杂志》2023年第10期878-882,共5页Ningxia Medical Journal

基　　金：宁夏自然科学基金资助项目(2022AAC03367)。

摘　　要：目的研究应用T细胞特异性慢病毒过表达白细胞介素10(IL-10)或siRNA片段抑制IL-10后对小鼠脾脏CD4+T细胞亚群的影响。方法磁珠分选试剂分离小鼠初始CD4+T细胞,通过IL-10过表达特异性慢病毒侵染CD4+T细胞,应用qRT-PCR检测IFN-γ、IL-4、IL-17A、Foxp3的mRNA表达量,应用流式细胞术分别检测IFN-γ、IL-4、IL-17A、Foxp3在CD4+T细胞中的分泌比率。应用siRNA干扰片段抑制IL-10,应用qRT-PCR检测IFN-γ、IL-4、IL-17A、Foxp3的mRNA表达量,应用流式细胞术分别检测IFN-γ、IL-4、IL-17A、Foxp3在CD4+T细胞中的分泌比率。结果过表达IL-10后,IFN-γ、IL-4、IL-17A mRNA水平明显下降(P<0.05),Foxp3 mRNA表达明显上升(P<0.05);IFN-γ、IL-4、IL-17A分泌量均明显降低(P<0.05),Foxp3分泌量明显增高(P<0.05)。IL-10抑制后,IFN-γ和IL-17A mRNA表达明显上升(P<0.05);IFN-γ和IL-17A分泌量均明显升高(P<0.05)。结论过表达IL-10后抑制Th1、Th2、Th17亚群中的IFN-γ、IL-4、IL-17A的表达,对Treg细胞Foxp3的分泌具有促进作用。IL-10抑制能明显促进IFN-γ、IL-17A的表达。Objective To study the effect of CD4+T cell subsets in mouse spleen after inhibiting IL-10 by using T cell specific lentiviruses overexpression of IL-10 or siRNA fragments respectively.Methods Magnetic bead sorting reagent was used to isolate initial CD4+T cells from mouse.IL-10 overexpression specific lentiviruses and Inhibition of IL-10 using siRNA interference fragments were used to infect CD4+T cells respectively.The mRNA expression levels of IFN-γ,IL-4,IL-17A and Foxp3 were tested by qRT-PCR,and the secretion ratios of IFN-γ,IL-4,IL-17A and Foxp3 were tested by flow cytometry.Results The mRNA levels of IFN-γ,IL-4,IL-17A were significantly decreased after IL-10 overexpression(P<0.05)and the expression of Foxp3 mRNA significantly increased(P<0.05).The secretion of IFN-γ,IL-4,IL-17A significantly decreased(P<0.05)and the secretion of Foxp3 significantly increased(P<0.05).After IL-10 inhibition,the mRNA levels of IFN-γand IL-17A significantly increased(P<0.05),and the secretion of IFN-γand IL-17A significantly increased(P<0.05).Conclusion Overexpression of IL-10 can inhibite the expression of IFN-γ,IL-4 and IL-17A,and significantly promote the expression of Foxp3.IL-10 inhibition can significantly promote the expression of IFN-γand IL-17A.

关 键 词：CD4+T细胞 IL-10 慢病毒过表达 SIRNA抑制 

分 类 号：R532.14[医药卫生—内科学]