Pitavastatin up-regulates the induction of iNOS through enhanced stabilization of its mRNA in pro-inflammatory cytokine-stimulated hepatocytes
Section snippets
Materials
Recombinant human IL-1β (2 × 107 units/mg protein) was provided by Otsuka Pharmaceutical Co. Ltd. (Tokushima, Japan). [γ-32P]Adenosine-5′-triphosphate (ATP, −222 TBq/mmol) and [α-32P]deoxycytidine-5′-triphosphate (dCTP, −111 TBq/mmol) were from DuPont-New England Nuclear Japan (Tokyo). Pitavastatin was provided by Kowa Co. Ltd. (Nagoya, Japan) and dissolved in Williams′ medium E (WE). All other chemicals were of reagent grade. Rats were kept at 22 °C under a 12 h light–dark cycle, and received food
Pitavastatin up-regulates the induction of iNOS in hepatocytes
The pro-inflammatory cytokine, IL-1β, stimulated the induction of iNOS gene expression, resulting in an increased NO production, in primary cultures of rat hepatocytes as reported previously [29], [32], [33], [34]. Pretreatment with pitavastatin (HMG-CoA reductase inhibitor, 50 μM) for 2 h or more (up to overnight) before the addition of IL-β markedly increased the production of NO with time (approximately 3- to 4-fold) compared with IL-β alone (Fig. 1a). Simultaneous addition of pitavastatin (50
Discussion
In this study, we found that an HMG-CoA reductase inhibitor, pitavastatin, significantly up-regulated the induction of iNOS gene expression, as stimulated by pro-inflammatory cytokine IL-1β in cultured hepatocytes, followed by an increased production of NO (Fig. 1, Fig. 2). However, pitavastatin had no effects on either the degradation of IκBα and IκBβ or the activation of NF-κB (Fig. 3). In contrast, pitavastatin time- and dose-dependently enhanced the induction of IL-1RI mRNA and protein (
Acknowledgments
This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Culture and Sports of Japan, and by grants from the Science Research Promotion Fund of the Japan Private School Promotion Foundation, and from Amino Up Chemical Co., Ltd. (Sapporo, Japan).
References (45)
- et al.
The evolving role of statins in the management of atherosclerosis
J. Am. Coll. Cardiol.
(2000) - et al.
Simvastatin enhances hepatic nitric oxide production and decreases the hepatic vascular tone in patients with cirrhosis
Gastroenterology
(2004) - et al.
Simvastatin attenuates expression of cytokine-inducible nitric-oxide synthase in embryonic cardiac myoblasts
J. Biol. Chem.
(2005) - et al.
Reduced nitric oxide production by endothelial cells in cirrhotic rat liver: endothelial dysfunction in portal hypertension
Gastroenterology
(1998) - et al.
Impaired endothelial nitric oxide synthase activity associated with enhanced caveolin binding in experimental cirrhosis in the rat
Gastroenterology
(1999) - et al.
Stimulation of glycogen degradation by prostaglandin E2 in primary cultured rat hepatocytes
Prostaglandins
(1993) Preparation of isolated rat liver cells
Methods Cell Biol.
(1976)- et al.
Analysis of nitrate, nitrite and [15N]nitrate in biological fluids
Anal. Biochem.
(1982) - et al.
Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction
Anal. Biochem.
(1987) - et al.
Cloning of inducible nitric oxide synthase in rat vascular smooth muscle cells
Biochem. Biophys. Res. Commun.
(1993)
An mRNA homologous to interleukin-1 receptor type I is expressed in cultured rat sympathetic ganglia
J. Neuroimmunol.
Glyceraldehyde-3-phosphate dehydrogenase mRNA is a major interleukin 2-induced transcript in a cloned T-helper lymphocyte
Gene
Characterization of the isoforms of MOVO zinc finger protein, a mouse homologue of Drosophila Ovo, as transcription factors
Gene
Vicinal dithiol-binding agent, phenylarsine oxide, inhibits iNOS gene expression at a step of NF-κB DNA binding in hepatocytes
J. Biol. Chem.
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding
Anal. Biochem.
Anti-inflammatory drug sodium salicylate inhibits nitric oxide formation induced by interleukin-1b at a translational step, but not a transcriptional step, in hepatocytes
Hepatology
Hypoxia and heat inhibit inducible nitric oxide synthase gene expression by different mechanisms in rat hepatocytes
Hepatology
Interleukin-1β markedly stimulates nitric oxide formation in the absence of other cytokines or lipopolysaccharide, in primary cultured rat hepatocytes, but not in Kupffer cells
Hepatology
Up-regulation of IL-1 receptor through PI3K/Akt is essential for the induction of iNOS gene expression in hepatocytes
J. Hepatol.
Regulation of the expression of inducible nitric oxide synthase
Eur. J. Pharmacol.
The polypyrimidine tract-binding protein (PTB) is involved in the post-transcriptional regulation of human inducible nitric oxide synthase expression
J. Biol. Chem.
Distinct roles of the IkappaB kinase alpha and beta subunits in liberating nuclear factor kappaB (NF-kappaB) from IkappaB and in phosphorylating the p65 subunit of NF-kappaB
J. Biol. Chem.
Cited by (23)
Anti-inflammatory effects of pitavastatin in interleukin-1β-induced SW982 human synovial cells
2017, International ImmunopharmacologyCitation Excerpt :Statins have been reported to enhance production of nitric oxide (NO), previous study has found that pitavastatin induces NO production through the Akt pathway in human umbilical vein endothelial cells [31]. Habara et al. demonstrated that the induction of inducible NO synthase (iNOS) was up-regulated by pitavastatin in hepatocytes [32]. Studies have reported that the iNOS, a sort of nitric oxide synthase, results in an increase in NO production in OA synoviocytes [33–34].
Natural antisense transcript-targeted regulation of inducible nitric oxide synthase mRNA levels
2013, Nitric Oxide - Biology and ChemistryCitation Excerpt :In rat hepatocytes treated with the proinflammatory cytokine interleukin-1β (IL-1β), rapid induction of iNOS asRNA is observed [4]. Insulin-like growth factor I (IGF-I), the anti-inflammatory drug dexamethasone, and the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) inhibitor pitavastatin all affect rat iNOS asRNA levels [9–11]. Additionally, iNOS asRNA is expressed in models of septic shock using rats treated with d-galactosamine and lipopolysaccharide (LPS) [9,12].
Aristolochic acid I suppressed iNOS gene expression and NF-κB activation in stimulated macrophage cells
2011, Toxicology LettersCitation Excerpt :A regulatory cis-element within the 3′-UTR of iNOS mRNA has been identified and is supposed to play an important role in iNOS mRNA stability during extended stimulation by LPS (Soderberg et al., 2007). Various compounds have been shown to act on the 3′-UTR of iNOS gene and thus to increase or to decrease the stability of iNOS mRNA in mouse hepatocytes or macrophages (Habara et al., 2008; Yoshida et al., 2008). In particular, a natural antisense transcript that can stabilize iNOS mRNA has been identified; the iNOS antisense transcript may interact with the iNOS mRNA 3′-UTR with the help of RNA binding proteins (Matsui et al., 2008).
Simvastatin-dependent up-regulation of heme oxygenase-1 via mRNA stabilization in human endothelial cells
2010, European Journal of Pharmaceutical SciencesRegulation of the expression of inducible nitric oxide synthase
2010, Nitric Oxide - Biology and ChemistryCobalt protoporphyrin inhibition of lipopolysaccharide or lipoteichoic acid-induced nitric oxide production via blocking c-Jun N-terminal kinase activation and nitric oxide enzyme activity
2009, Chemico-Biological InteractionsCitation Excerpt :In large amounts, NO is considered a deleterious molecule in the process of inflammation and sepsis. After exposure to endogenous and exogenous stimulators, inducible NO synthase (iNOS) is quantitatively induced in various cells such as macrophages, smooth muscle cells, and hepatocytes to trigger several deleterious cellular responses [9–11]. Lipopolysaccharide (LPS) is a major component of gram-negative bacteria, and promotes the secretion of inflammatory cytokines and induces iNOS gene expression via binding to the CD14/Toll-like receptor (TLR) 4 in macrophages [11,12].