TLR4 and MD2 antisense genes suppress activation of alveolar typeⅡ epithelial cells induced by lipopolysaccharide / 第三军医大学学报

ABSTRACT

Objective To observe effects of the antisense genes of Toll-like receptor 4 (TLR4 ) and myeloid differentiation protein-2 (MD2) on the activation of NF-?B in alveolar typeⅡepithelial cells (ATⅡcells) induced by lipopolysaccharide (LPS). Methods Cultured ATⅡcells were randomized to normal cells (control) group,LPS group,LPS+empty vector group,LPS+TLR4 antisense gene group,LPS+MD2 antisense gene group,LPS+TLR4-MD2 antisense gene group. The expressions of TLR4 and MD2 mRNA and protein in ATⅡcells were detected by Northern blotting and Western blotting respectively. The activity of NF-?B in ATⅡcells was measured with electrophoretic mobility shift assay (EMSA). The TNF-? and IL-6 concentrations in the supernatant of cultured ATⅡcells were tested with ELISA.Results Compared with control groups,the expressions of TLR4 and MD2 mRNA and protein,the NF-?? activity,the levels of TNF-? and IL-6 were increased remarkably in LPS group and LPS+empty vector group (P