γ促分泌酶抑制剂MW167对毛细支气管炎患儿外周血单个核细胞中Th17细胞比例的影响

摘要

目的 观察γ促分泌酶抑制剂MW167对呼吸道合胞病毒(RSV)毛细支气管炎患儿外周血单个核细胞(PBMC)中Th17细胞比例、视黄酸受体相关孤核受体γt(RORγt)mRNA表达及细胞上清液中IL-17水平的影响.方法 选择RSV毛细支气管炎30例患儿作为观察组,25例健康体检儿作为对照组,抽取患儿外周静脉血,分离PBMC.将对照组儿童外周血分离所得的PBMC作为正常组,观察组患儿外周血分离所得的PBMC作为RSV组、MW167组,MW167组加入MW167,培养48 h.采用流式细胞术检测三组Th17细胞比例,ELISA检测细胞培养上清液中IL-17水平,实时荧光定量PCR检测细胞中的RORγt mRNA表达.结果 正常组、RSV组、MW167组PBMC中Th17细胞比例分别为0.49％±0.09％、1.56％±0.77％、0.84％±0.23％,RSV组与正常组相比P＜0.01,MW167组与RSV组相比P＜0.05.正常组、RSV组、MW167组PBMC细胞培养上清液中IIL-17水平分别为(12.18±2.97)、(20.88 ±4.10)、(12.91 ±4.28) pg/mL,RSV组与正常组相比,MW167组与RSV组相比,P均＜0.05.正常组、RSV组、MW167组PBMC中RORγt mRNA相对表达量分别为1.0、5.72 ±0.53、1.82 ±0.21,RSV组与正常组相比,MW167组与RSV组相比,P均＜0.05.结论 γ促分泌酶抑制剂MW167可降低RSV毛细支气管炎患儿PBMC中RORγt mRNA表达、Th17细胞比例及细胞培养上清液中IL-17水平.%Objective To observe the effects of γ-secretase inhibitor MW167 on the proportion of Th17 cells,the expression of retinoic acid receptor-related orphan nuclear receptor γt (RORγt) mRNA,and the level of interleukin-17 (IL-17) in the cell supematant of peripheral blood mononuclear cells (PBMCs) of children with respiratory syncytial virus (RSV) bronchiolitics.Methods Thirty children with RSV bronchiolitics were selected as the observation group and 25 healthy children as the control group.Peripheral blood was drawn from children and PBMCs were isolated.PBMCs which isolated from healthy children were taken as the normal group,and PBMCs from the observation group were then divided into the RSV group and MW167 group.Cells in the MW167 group were added with MW167 and were cultured for 48 h.The proportion of Th17 cells in the three groups was detected by flow cytometry.The level of IL-17 in the cell culture supernatants was detected by ELISA.The mRNA expression of RORγt was detected by real-time fluorescence quantitative PCR.Results The proportion of Th17 cells in PBMCs of the normal group,RSV group,and MW167 group was 0.49％ ± 0.09％,1.56％ ±0.77％,and 0.84％ ±0.23％,and significant difference was found between the RSV group and the normal group,as well as between the M W167 group and the RSV group (P ＜ 0.01 or P ＜ 0.05).The levels of IL-17 in the PBMC culture supernatants of the normal group,RSV group,and MW167 group were (12.18 ±2.97),(20.88 ±4.10) and (12.91 ±4.28) pg/mL,respectively;significant difference was found between the RSV group and the normal group,and between the MW167 group and the RSV group (both P ＜ 0.05).The relative expression levels of RORγt mRNA in PBMCs of the normal group,RSV group,and MW167 group were 1.0,5.72 ±0.53,and 1.82 ±0.21,respectively;significant difference was found between the RSV group and the normal group,and between the MW167 group and the RSV group (both P ＜ 0.05).Conclusion The γ-secretase inhibitor MW167 can reduce the expression of RORγt mRNA,the proportion of Th17 cells,and the level of IL-17 in PBMCs of children with RSV bronchiolitis