Hepatic ischemia/reperfusion upregulates the susceptibility of hepatocytes to confer the induction of inducible nitric oxide synthase gene expression.
During hepatic ischemia/reperfusion (I/R), proinflammatory cytokines such as tumor necrosis factor alpha and interleukin (IL) 1beta stimulate the induction of inducible nitric oxide synthase (iNOS) in hepatocytes, followed by massive production of nitric oxide. We hypothesized that I/R upregulated the susceptibility of hepatocytes to confer the induction of iNOS gene expression. This study was designed to investigate whether cell susceptibility occurs in response to I/R and to delineate the mechanisms underlying the susceptibility. Hepatocytes were isolated from rats with hepatic I/R or sham, cultured, and treated with IL-1beta. The iNOS induction and its signal including inhibitor kappaB (IkappaB) kinase/nuclear factor kappaB (NF-kappaB) and Akt/type 1 interleukin 1 receptor (IL-1R1) were analyzed. Hepatocytes isolated from rats with I/R markedly increased the production of nitric oxide when stimulated by IL-1beta as compared with sham control. Ischemia/R also increased the levels of iNOS protein and its messenger RNA. Furthermore, I/R enhanced the activation of transcription factor NF-kappaB and the transactivation of iNOS promoter. However, I/R had no effects on the degradation of IkappaB and the nuclear translocation of p65 subunit of NF-kappaB. In contrast, I/R increased the phosphorylation of Akt and the upregulation of IL-1R1 induction, which is essential signal for the transcriptional activation of iNOS in addition to IkappaB kinase/NF-kappaB. These results demonstrate that I/R may augment hepatocyte susceptibility for the induction of iNOS gene expression through the enhancement of IL-1R1.
Yanagida
Hidesuke
H
The Department of Surgery, Kansai Medical University, Osaka 570-8506, Japan.
Kaibori
Masaki
M
Yoshida
Hideyuki
H
Habara
Kozo
K
Yamada
Masanori
M
Kamiyama
Yasuo
Y
Okumura
Tadayoshi
T
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000201:enzymology* / Q000473:pathology; D020672:I-kappa B Proteins / Q000378:metabolism; D007375:Interleukin-1 / Q000494:pharmacology; D007511:Ischemia / Q000201:enzymology*; D008099:Liver / Q000098:blood supply* / Q000201:enzymology; D008297:Male; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics* / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017472:Receptors, Interleukin-1 / Q000187:drug effects / Q000378:metabolism; D053573:Receptors, Interleukin-1 Type I; D015424:Reperfusion; D015427:Reperfusion Injury / Q000201:enzymology*; D015854:Up-Regulation
D020672:I-kappa B Proteins; D007375:Interleukin-1; D016328:NF-kappa B; D017472:Receptors, Interleukin-1; D053573:Receptors, Interleukin-1 Type I; D052247:Nitric Oxide Synthase Type II; D051058:Oncogene Protein v-akt