TY - JOUR
T1 - A DNA microarray study of nitric oxide-induced genes in mouse hepatocytes
T2 - Implications for hepatic heme oxygenase-1 expression in ischemia/reperfusion
AU - Zamora, Ruben
AU - Vodovotz, Yoram
AU - Aulak, Kulwant S.
AU - Kim, Peter K.M.
AU - Kane, John M.
AU - Alarcon, Louis
AU - Stuehr, Dennis J.
AU - Billiar, Timothy R.
N1 - Funding Information:
This work was supported by Grants GM 44100 (to T.R. Billiar), CA 53914, and GM 51491 (to D.J. Stuehr) from the National Institutes of Health. Ruben Zamora was supported by a Research Fellowship from the Belgian American Educational Foundation (BAEF). The authors thank Ray Ganster (Department of Surgery, University of Pittsburgh) for critical reading of the manuscript and Debra L. Williams and Binnie Betten for skillful assistance with isolation of hepatocytes, cell transfection, and Northern blotting techniques.
PY - 2002
Y1 - 2002
N2 - Nitric oxide (NO) can modulate numerous genes directly; however, some genes may be modulated only in the presence of the inflammatory stimuli that increase the expression of the inducible nitric oxide synthase (iNOS). One method by which to examine changes in NO-mediated gene expression is to carry out a gene array analysis on NO-naïve cells. Herein, we report a gene array analysis on mRNA from iNOS-null (iNOS-/-) mouse hepatocytes harvested from mice exposed to NO by infection with an adenovirus expressing human iNOS (Ad-iNOS). Of the 6500 genes on this array, only approximately 200 were modulated either up or down by the increased iNOS activity according to our criteria for significance. Several clearly defined families of genes were modulated, including genes coding for proinflammatory transcription factors, cytokines, cytokine receptors, proteins associated with cell proliferation and cellular energetics, as well as proteins involved in apoptosis. Our results suggest that iNOS has a generally anti-inflammatory and anti-apoptotic role in hepatocytes but also acts to suppress proliferation and protein synthesis. The expression of iNOS results in increased expression of stress-related proteins, including heme oxygenase-1 (HO-1). We used HO-1 to confirm that a significant change identified by an analysis could be demonstrated as significant in cells and tissues. The elevation of HO-1 was confirmed at the protein level in hepatocytes in vitro. Furthermore, iNOS-/- mice experienced greatly increased liver injury subsequent to intestinal ischemia/reperfusion injury, associated with an inability to upregulate HO-1. This is the first study to address the global gene changes induced by iNOS in any cell type, and the findings presented herein may have clinical relevance for conditions such as septic or hemorrhagic shock in which hepatocytes, NO, and HO-1 play a crucial role.
AB - Nitric oxide (NO) can modulate numerous genes directly; however, some genes may be modulated only in the presence of the inflammatory stimuli that increase the expression of the inducible nitric oxide synthase (iNOS). One method by which to examine changes in NO-mediated gene expression is to carry out a gene array analysis on NO-naïve cells. Herein, we report a gene array analysis on mRNA from iNOS-null (iNOS-/-) mouse hepatocytes harvested from mice exposed to NO by infection with an adenovirus expressing human iNOS (Ad-iNOS). Of the 6500 genes on this array, only approximately 200 were modulated either up or down by the increased iNOS activity according to our criteria for significance. Several clearly defined families of genes were modulated, including genes coding for proinflammatory transcription factors, cytokines, cytokine receptors, proteins associated with cell proliferation and cellular energetics, as well as proteins involved in apoptosis. Our results suggest that iNOS has a generally anti-inflammatory and anti-apoptotic role in hepatocytes but also acts to suppress proliferation and protein synthesis. The expression of iNOS results in increased expression of stress-related proteins, including heme oxygenase-1 (HO-1). We used HO-1 to confirm that a significant change identified by an analysis could be demonstrated as significant in cells and tissues. The elevation of HO-1 was confirmed at the protein level in hepatocytes in vitro. Furthermore, iNOS-/- mice experienced greatly increased liver injury subsequent to intestinal ischemia/reperfusion injury, associated with an inability to upregulate HO-1. This is the first study to address the global gene changes induced by iNOS in any cell type, and the findings presented herein may have clinical relevance for conditions such as septic or hemorrhagic shock in which hepatocytes, NO, and HO-1 play a crucial role.
UR - http://www.scopus.com/inward/record.url?scp=1842834977&partnerID=8YFLogxK
U2 - 10.1016/S1089-8603(02)00104-0
DO - 10.1016/S1089-8603(02)00104-0
M3 - Article
C2 - 12381414
AN - SCOPUS:1842834977
SN - 1089-8603
VL - 7
SP - 165
EP - 186
JO - Nitric Oxide - Biology and Chemistry
JF - Nitric Oxide - Biology and Chemistry
IS - 3
ER -