The modulating effects of transfected IkappaBalpha gene on the NF-kappaB activity of hepatic tissue in scalded rats.
- Author:
Minghai WANG
1
;
Guangping TAI
;
Yaping ZHANG
;
Guangxia XIAO
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Animals; Burns; genetics; metabolism; therapy; Genetic Therapy; methods; Genetic Vectors; genetics; I-kappa B Proteins; genetics; metabolism; Interleukin-1; genetics; Liver; metabolism; NF-KappaB Inhibitor alpha; NF-kappa B; metabolism; RNA, Messenger; genetics; metabolism; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; Transfection; methods; Tumor Necrosis Factor-alpha; genetics
- From: Chinese Journal of Burns 2002;18(1):42-44
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the modulating effects mediated by recombinant IkappaBalpha gene by adenovirus vector transfection on the NF-kappaB activity of hepatic tissue in scalded rats.
METHODSAfter being primed by recombinant defect adenovirus (AdIkappaBalpha) vector, the rats were scalded. The hepatic tissue was harvested at different time points and the nuclear protein was extracted and reacted with [r(-23) P] ATP labelled NF-kB specific probe. NF-Kb/DNA combining activity was determined with electrophoretic mobility shifty assay (EMSA). The total RNA in hepatic tissue was extracted and the expressions of IL-1beta and TNFalpha mRNA were determined with RT-PCR.
RESULTSWhen compared to those in normal control, the NF-kappaB/DNA combining activity increased significantly in scalded rats at half an hour after scalding, and it lasted for 24 PBDs with rather strong activity. But in AdIkappaBalpha priming group, the rat NF-kappaB/DNA binding activity was slightly higher than that of normal control group, but was obviously lower than that of scalding group.
CONCLUSIONThe intracellular NF-kappaB in hepatic tissue was activated rapidly after that the rats were severely scalded, and the expression of IL-1beta and TNFalpha was enhanced significantly simultaneously. Priming of AdIkappaBalpha could evidently inhibit the activation of hepatic tissue NF-kB in rats injured by severe scalding, and down-regulated the expressions of IL-1beta and TNFalpha.
