O²-Vinyl 1-(Pyrrolidin-1-yl)diazen-1-ium-1,2-diolate Protection Againstd-Galactosamine/Endotoxin-Induced Hepatotoxicity in Mice: Genomic Analysis Using Microarrays

Abstract

O 2 -Vinyl 1-(pyrrolidin-1-yl)diazen-1-ium-1,2-diolate (V-PYRRO/ NO), a liver-selective nitric oxide (NO)-donating prodrug, is metabolized by hepatic enzymes to release NO within the liver. This study was undertaken to examine the effects of V-PYRRO/NO on D-galactosamine/lipopolysaccharide (GlaN/ LPS)-induced liver injury in mice. Mice were given injections of V-PYRRO/NO (10 mg/kg, s.c. at 2-h intervals) before and after GlaN/LPS (700 mg/30 g/kg, i.p.). V-PYRRO/NO administration dramatically reduced GlaN/LPS-induced hepatotoxicity, as evidenced by reduced serum alanine aminotransferase activity and improved pathology. To examine the mechanisms of the protection, cDNA microarray was performed to profile the gene expression pattern in livers of mice treated with GlaN/LPS, GlaN/LPS plus V-PYRRO/NO, or controls. V-PYRRO/NO administration greatly ameliorated GlaN/LPS-induced alterations in the expression of genes encoding the stress response, DNA damage/repair response, and drug-metabolizing enzymes in accordance with hepatoprotection. Gel shift assay and Western blot analysis supported microarray results, showing that V-PYRRO/NO suppressed GlaN/LPS-induced activation of nuclear factor-B and GlaN/LPS-induced increases in caspase-1, caspase-8, tumor necrosis factor receptor 1 (TNFR1)-associated death domain, and TNF-related apoptosis-inducing ligand. Immunohistochemical analysis further revealed that GlaN/LPS-induced activation of TNFR1, caspase-3, and hepatocellular apoptosis was ameliorated by V-PYRRO/NO treatment. GlaN/LPS-induced elevation of hepatic caspase-3 activity was diminished by V-PYRRO/NO treatment. In addition, V-PYRRO/NO alone suppressed the basal expression of genes encoding inducible NO synthase and TNF-␣-related components, as revealed by mouse 1.2 array. In summary, this study demonstrates that the liver-selective NO donor, V-PYRRO/NO, is effective in blocking GlaN/LPS-induced hepatotoxicity in mice, and that this protection appears to involve, at least in part, the suppression of the TNF-␣-mediated cell death pathways.

The nitric oxide donor, V-PYRRO/NO, protects against acetaminophen-induced hepatotoxicity in mice

“…12 The structure of V-PYRRO/NO and the mode of NO release have been published previously. 12,13,15 Osmotic pumps (200-L capacity, 8 L/h) were purchased from Alzet (Palo Alto, CA). Acetaminophen was purchased from Sigma Chemical Co. (St. Louis, MO).…”

“…13 The systemic effects of V-PYRRO/NO have been shown to be minimal. [12][13][14][15] V-PYRRO/NO has been shown to protect against tumor necrosis factor ␣ (TNF-␣)-induced hepatic apoptosis, 12,16,17 monocrotaline-induced hepatic sinusoid injury, 18 liver damage from ischemia reperfusion, 14 D-galactosame/endotoxin-induced hepatotoxicity, 15 and bile duct ligation-induced portal hypertension and fibrosis. 19 The role of NO in acetaminophen hepatotoxicity is still an issue of debate, apparently due to the dual effects of NO.…”

The nitric oxide prodrug, V-PYRRO/NO, mitigates arsenic-induced liver cell toxicity and apoptosis

“…V-PYRRO/NO mitigates the hepatotoxicity of various compounds in vivo and in vitro [10][11][12]. For instance, V-PYRRO/NO can protect against LPS-induced liver injury in mice, an effect that is linked to suppression of apoptosis and NF-κB expression [11]. V-PYRRO/NO also reduces TNF-α-induced liver injury associated with reduced hepatocellular apoptosis [9].…”

“…O 2 -vinyl 1-(pyrrolidin-1-yl)diazen-1-ium-1,2-diolate (V-PYRRO/NO) is a stable diazeniumdiolate, which can circulate freely in the body until it is metabolized to release NO by cytochrome P450 in the liver, providing a significant level of local hepatic NO [9]. V-PYRRO/NO mitigates the hepatotoxicity of various compounds in vivo and in vitro [10][11][12]. For instance, V-PYRRO/NO can protect against LPS-induced liver injury in mice, an effect that is linked to suppression of apoptosis and NF-κB expression [11].…”

The morphological and immunocytochemical evaluation of primary rat hepatocytes undergoing spontaneous cell death: Modulation by the nitric oxide donor S-nitroso-N-acetylpenicillamine

“…Another important aspect of NO interaction with cellular functions is its effect on apoptosis. There are studies, which indicate that NO possesses antiapoptotic effects in the liver as NO improves hepatocyte survival after liver injury caused by apoptosis inducing agents 9,10 . In the present study we evaluated the effect of the NO donor, S-nitroso-N-acetylpenicillamine (SNAP), on the viability and several other functional parameters of primary rat hepatocytes.…”