Effect of lentivirus expressing TLR4 shRNA on function of rat alveolar macrophage
- VernacularTitle:慢病毒介导shRNA特异性沉默TLR4基因对大鼠肺泡巨噬细胞功能的影响
- Author:
Jian LIU
;
Huimin FAN
;
Zhongmin LIU
- Publication Type:Journal Article
- Keywords:
Toll-like receptor 4;
RNA interference;
lentivirus;
interleukin-1?;
interleukin-6
- From:
Academic Journal of Second Military Medical University
1985;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the replication-incompetent lentivirus expressing rat TLR4 shRNA,and to observe its inhibitory effect on TLR4 expression in alveolar macrophage and on lipopolysaccharide (LPS)-induced release of IL-1?,IL-6.Methods We designed and constructed four shRNA expressing plasmids with silencing effect,then they were co-transfected into HEK293T cells with the TLR4 eukaryotic vector constructed previously;the best shRNA was selected to be packed into lentivirus;and the titer was determined.The packed lentvirus was used to infect the alveolar macrophage(NR8383) in presence of LPS,and the LPS-induced IL1?,IL-6 expression was examined by ELISA.Results The best shRNA was successfully screened out and was correctly inserted into the lentivirus.The titer of the recombinant lentivirus was 2.0 ? 106 TU/ml.The LPS-induced expression of IL-1?,IL-6 in the alveolar macrophages was greatly reduced after virus infection (P
