TY - JOUR
T1 - Induction of Nitric Oxide Synthesis and Its Reactions in Cultured Human and Rat Hepatocytes Stimulated with Cytokines Plus LPS
AU - Nussler, Andreas K.
AU - Geller, David A.
AU - Sweetland, Michael A.
AU - Di Silvio, Mauricio
AU - Billiar, Timothy R.
AU - Madariaga, Juan B.
AU - Simmons, Ricahrd L.
AU - Lancaster, Jack L.
PY - 1993/7/30
Y1 - 1993/7/30
N2 - We have examined the time course of appearance of mRNA for nitric oxide synthase (NOS), intracellular nonheme iron-nitrosyl complexes (NHFeNO, detected by EPR spectroscopy), and rates of medium appearance of NO2- + NO3- in cultured rat and human hepatocytes stimulated with a combination of cytokines (TNF-α, IFN-γ, IL-1β) and LPS. In both cells types, NOS mRNA precedes NHFeNO formation which in turn precedes maximum rates of NO2- + NO3- (NOx) formation. This profile occurs earlier in human hepatocytes than rat hepatocytes and the appearance of NOS mRNA is also more transient. These results indicate that (1) NOS is stable intracellularly (peak NOx production occurs substantially after peak mRNA levels), (2) intracellular iron is an early target (preceding maximum NOx production) for NO in both human and rat hepatocytes, and (3) decline in NHFeNO in the face of maximum NOx production indicates the presence of a “repair” or “removal” mechanism for these intracellular iron-nitrosyl complexes.
AB - We have examined the time course of appearance of mRNA for nitric oxide synthase (NOS), intracellular nonheme iron-nitrosyl complexes (NHFeNO, detected by EPR spectroscopy), and rates of medium appearance of NO2- + NO3- in cultured rat and human hepatocytes stimulated with a combination of cytokines (TNF-α, IFN-γ, IL-1β) and LPS. In both cells types, NOS mRNA precedes NHFeNO formation which in turn precedes maximum rates of NO2- + NO3- (NOx) formation. This profile occurs earlier in human hepatocytes than rat hepatocytes and the appearance of NOS mRNA is also more transient. These results indicate that (1) NOS is stable intracellularly (peak NOx production occurs substantially after peak mRNA levels), (2) intracellular iron is an early target (preceding maximum NOx production) for NO in both human and rat hepatocytes, and (3) decline in NHFeNO in the face of maximum NOx production indicates the presence of a “repair” or “removal” mechanism for these intracellular iron-nitrosyl complexes.
UR - http://www.scopus.com/inward/record.url?scp=0027162258&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1993.1896
DO - 10.1006/bbrc.1993.1896
M3 - Article
C2 - 7688228
AN - SCOPUS:0027162258
SN - 0006-291X
VL - 194
SP - 826
EP - 835
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -