Inhibitory effect of special siRNA targeting toll-like-receptor 2 gene on the inflammatory factor expression of alveolar epithelial cells under hyperoxia-induced oxidative stress
Huang Dong
Fang Fang
Xu Feng
Abstract：Objective To study the function of toll-like receptor 2 (TLR2) gene and to observe if the intracellular reactive oxygen species (ROS) can activate the signal transduction pathway of TLR2-nuclear factor (NF)-KB-inflammatory mediators resulting in inflammatory response during hyperoxia-induced acute lung injury.Methods The A549 was cultured in six-well plates and randomly divided into group N (air group),group H (hyperoxia group),group T(hyperoxia + TLR2-siRNA transfected group) and group C(hyperoxia + control-siRNA transfected group).The siRNA was transfected into A549 cells mediated by lipofectamine 2000.Group H,group T and group C were cultured in hyperoxia condition for 6 h.The expression of FAM was detected by flow eytometry to observe the efficiency of transfection.Real-time fluorescence polymerase chain reaction (Real-time PCR) method was used to detect the level of mRNA of TLR2.TLR2 protein expression was observed by flow cytometry.The activity of NF-κB in A549 cells was measured with electrophoretic mobility shift assay (EMSA) and enzyme-linked immunosorbent assay(ELISA) was used to test the level of IL-6 and IL-8 in the supematants.Results The expression of FAM was (92.3 ±9.6)％ after transfection.Compared with group N,the TLR2 mRNA,TLR2 protein level,activity of NF-κB and the IL-6 and IL-8 quantity in group H and group C increased after the hyperoxia treatment[(1.68 ±0.02),(1.52 ±0.06) vs (1.26 ±0.02) ;(9.1 ±0.8),(7.8 ± 1.4) vs (5.9 ±2.8) ;(484 ± 64),(378 ±38) vs (46 ±3)] (P ＜0.05).While the expression of the TLR2 mRNA(0.87 ±0.06),TLR2 protein level(2.8 ± 0.47),activity of NF-κB and the IL-6 and IL-8 quantity in the group T were decreased compared with group H and group C [L-6:(48.5 ± 6.3) ng/L vs (66.7 ± 7.8),(59.6 ± 6.5) ng/L,IL-8:(125 ± 19) ng/L vs (773 ± 96),(971 ± 149)ng/L] (all P ＜ 0.05).The group T had a significant difference from the group N regarding the expression of detection indexes (P ＜0.05).Conclusion The siRNA targeting TLR2 mRNA can inhibit the inflammatory reaction.
Keywords：Small interference RNAHyperoxiaToll-like receptor 2Alveolar epithelial cells
Publication Date：2014-01-01
Online Publishing Date：2026-09-14(First online date of this platform, not the publication date of the document)
Pages：5( 23-27 )