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In vitro and in vivo expression of inducible nitric oxide synthase during experimental endotoxemia: Involvement of other cytokines
Author(s) -
Aono Keiya,
Isobe Kenichi,
Kiuchi Kazutoshi,
Fan Zouheng,
Ito Masafumi,
Takeuchi Akihide,
Miyachi Masahiko,
Nakashima Izumi,
Nimura Yuji
Publication year - 1997
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19970601)65:3<349::aid-jcb5>3.0.co;2-s
Subject(s) - nitric oxide synthase , in vivo , in vitro , nitric oxide , chemistry , atp synthase , pharmacology , microbiology and biotechnology , medicine , biology , biochemistry , enzyme , organic chemistry
In this study, we investigated the expression of genes for inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF‐α), interleukin 1β (IL‐1β), interleukin 6 (IL‐6) of Kupffer cells in the presence of lipopolysaccharide (LPS), and the tissue expression of iNOS in a rat liver after LPS injection at various time intervals. The effects of L‐N G ‐nitroarginine‐methyl‐esther HCI (L‐NAMF), a NO inhibitor, also were examined. The mRNA transcripts of TNF‐α IL‐1β and IL‐6 were rapidly detected no more than at 1 h after LPS stimulation, whereas the iNOS transcript was detectable from 3 h after LPS stimulation and maximally increased at 12 h. This fact suggested that these early induced cytokines were related to expression of iNOS. Using an anti‐iNOS antiserum raised against recombinant iNOS protein, immunohistochemical analysis was made to reveal kinetics of NO producing cells. The cells immunoreactive for iNOS appeared at 6 h post‐LPS injection in the sinusoids of the liver. By structural and immunohistochemical studies, almost all iNOS positive cells were identified as Kupffer cells and endothelial cells. The number of cells immunoreactive for iNOS increased until 12 h post‐LPS injection. At 24 h after LPS injection, iNOS positive cells were restricted to the foci of spotty necrosis. Hepatic injury measured by released enzymes was increased by pretreatment of L‐NAME before LPS injection. J. Cell. Biochem. 65:349–358. © 1997 Wiley‐Liss, Inc.