The effect of siRNA inhibited tyrosine protein kinase Lck on the function of T cells in asthmatic mice
10.3760/ema.j.issn.1008-1372.2011.10.008
- VernacularTitle:SiRNA干扰靶向抑制酪氨酸蛋白激酶Lck对哮喘小鼠T细胞功能的影响
- Author:
Qiaoying JI
;
Shuangyan FANG
;
Caimin SHU
;
Qiongfang YANG
;
Dongli SONG
;
Yonghua ZHENG
- Publication Type:Journal Article
- Keywords:
RNA,small interfering;
Lymphocyte specific protein tyrosine kinase p56 (lck);
Asthma/IM;
Cytokines/ME
- From:
Journal of Chinese Physician
2011;13(10):1323-1326
- CountryChina
- Language:Chinese
-
Abstract:
Objective Using the technology of siRNA to inhibit gene expression of T cells'nonreceptor tyrosine protein kinase Lck in asthmatic mice,and to study the effect of siRNA inhibited Lck to the function of T cells in asthmatic mice.Methods The 21 - 23 bp RNA fragments of mouse T cell Lck were made by chemosynthesis.INTERFERinTMsiRNA Transfection Reagent was used as transfection reagent to transfect the siRNA into the spleen T cells of asthmatic mice for 48 hours.Then T cells were mixed with bone marrow dendritic cells (DC) of asthmatic mice for another 48 hours.Cell culture suspension was collected and the level of IL-4,IL-13,IL-2,INF-γ were detected with respondent ELISA kits; Western Blot was used to identify if the expression of Lck was blocked.Results The expression of Lck in T cells almost could not be detected in siRNA interference group.The levels of IL-4 and IL-13 in siRNA interference group( 10.19 ± 1.66,12.34 ±0.79) were lower than no-siRNA interference(28.06 ±2.88,27.87 ± 1.61 )and control group ( 22.07 ± 2.5 1,20.47 ± 2.37 ),and the difference was statistical significant ( P ＜0.01 ).Conclusions Special siRNA could block the expression of special gene,and Lck specific siRNA could block the activation and differentiation of T cells and reduce the secretion of inflammatory cytokines in asthmatic mice.
