[The modulating effects of transfected IkappaBalpha gene on the NF-kappaB activity of hepatic tissue in scalded rats].
OBJECTIVE
To explore the modulating effects mediated by recombinant IkappaBalpha gene by adenovirus vector transfection on the NF-kappaB activity of hepatic tissue in scalded rats.


METHODS
After 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.


RESULTS
When 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.


CONCLUSIONS
The 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.
Wang
Minghai
M
Institute of Burn Research, Southwestern Hospital, The Third Military Medical University, Chonqing 400038 P.R. China.
Tai
Guangping
G
Zhang
Yaping
Y
Xiao
Guangxia
G
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002056:Burns / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D020672:I-kappa B Proteins / Q000235:genetics* / Q000378:metabolism; D007375:Interleukin-1 / Q000235:genetics; D008099:Liver / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D020672:I-kappa B Proteins; D007375:Interleukin-1; D016328:NF-kappa B; C000606206:Nfkbia protein, rat; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha