中国当代儿科杂志2013，Vol.15Issue(4)：302-307,6.DOI:10.7499/j.issn.1008-8830.2013.04.014
短发夹RNA沉默Tim-3分子表达对哮喘小鼠外周血Th1和Th17细胞的影响
Small hairpin RNA silencing Tim-3 affects peripheral blood Th1 and Th17 cells differentiation in asthmatic mice
摘要
Abstract
Objective To investigate the effects of down-regulating Tim-3 gene in the peripheral blood mononuclear cells (PBMCs) of an asthmatic mouse model by short hairpin RNA( shRNA) and to explore the effect of Tim-3 on Thl and Thl7 cell differentiation. Methods An asthmatic murine model was established by ovalbumin sensitization and challenge. PBMCs were isolated from asthmatic mice and transfected by shRNA targeting Tim-3 gene. The mRNA and protein expressions of Tim-3 were detected by quantitative PCR and Western blot. Flow cytometry analysis was performed to determine the levels of Thl and Th17, and ELISA was performed to determine concentrations of IFN-γ, IL4 and IL-17 in the supernatant. Results Tim-3 mRNA expression in PBMCs was significantly increased in asthmatic mice. The mRNA and protein expression of Tim-3 decreased significantly in the shRNA group. Compared with the negative groups, Thl cell levels increased and Thl7 cell levels decreased significantly in the asthmatic groups after Tim-3 shRNA interference. In the Tim-3 shRNA interference groups concentrations of IFN-γ increased significantly while IL-17 decreased significantly. Conclusions Specific Tim-3 shRNA effectively silences the expression of Tim-3 and change in Tim-3 expression could affect T cell differentiation.关键词
哮喘/Tim-3/RNA沉默/T细胞/小鼠Key words
Asthma/ Tim-3/ RNA interference/ T helper cells/ Mice引用本文复制引用
陆小霞,徐佳莉,董宗祈,陈鹏,王莹..短发夹RNA沉默Tim-3分子表达对哮喘小鼠外周血Th1和Th17细胞的影响[J].中国当代儿科杂志,2013,15(4):302-307,6.基金项目
国家自然科学基金青年科学基金项目(项目编号:81200006) （项目编号:81200006）
吴阶平医学基金(项目编号:320.6750.1285) （项目编号:320.6750.1285）
湖北省自然科学基金(项目编号:2010CDB08804) （项目编号:2010CDB08804）
武汉市卫生局科研项目(项目编号:WX11B10). （项目编号:WX11B10）
