title: Withaferin-A Reduces Acetaminophen-Induced Liver Injury in Mice.
abstract: Withaferin-A (WA) has anti-oxidant activities however, its therapeutic potential in acetaminophen (APAP) hepatotoxicity is unknown. We performed a proof-of-concept study to assess the therapeutic potential of WA in a mouse model that mimics APAP-induced liver injury (AILI) in humans. Overnight fasted C57BL/6NTac (5-6 wk. old) male mice received 200 mg/kg APAP intraperitoneally (i.p.). After 1 h mice were treated with 40 mg/kg WA or vehicle i.p., and euthanized 4 and 16 h later; their livers were harvested and serum collected for analysis. At 4 h, compared to vehicle-treated mice, WA-treated mice had reduced serum ALT levels, hepatocyte necrosis and intrahepatic hemorrhage. All APAP-treated mice had reduced hepatic glutathione (GSH) levels however, reduction in GSH was lower in WA-treated when compared to vehicle-treated mice. Compared to vehicle-treated mice, livers from WA-treated mice had reduced APAP-induced JNK activation, mitochondrial Bax translocation, and nitrotyrosine generation. Compared to vehicle-treated mice, WA-treated mice had increased hepatic up-regulation of Nrf2, Gclc and Nqo1, and down-regulation of Il-6 and Il-1β. The hepatoprotective effect of WA persisted at 16 h. Compared to vehicle-treated mice, WA-treated mice had reduced hepatocyte necrosis and hepatic expression of Il-6, Tnf-α and Il-1β, increased hepatic Gclc and Nqo1 expression and GSH levels, and reduced lipid peroxidation. Finally, in AML12 hepatocytes, WA reduced H₂O₂-induced oxidative stress and necrosis by preventing GSH depletion. Collectively, these data show mechanisms whereby WA reduces necrotic hepatocyte injury, and demonstrate that WA has therapeutic potential in AILI.
authors.affiliation: Section of Gastroenterology and Hepatology, Digestive Health Center, Georgia Regents University, Augusta, GA, 30912, USA.
authors.forename: Ravirajsinh N
authors.initials: RN
authors.lastname: Jadeja
authors.affiliation: Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, Maryland, 21201, USA.
authors.forename: Nathalie H
authors.initials: NH
authors.lastname: Urrunaga
authors.affiliation: University of Maryland School of Medicine, Baltimore, MD, 21201, USA.
authors.forename: Suchismita
authors.initials: S
authors.lastname: Dash
authors.affiliation: Section of Gastroenterology and Hepatology, Digestive Health Center, Georgia Regents University, Augusta, GA, 30912, USA. Electronic address: skhurana@gru.edu.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Khurana
authors.affiliation: Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, Maryland, 21201, USA.
authors.forename: Neeraj Kumar
authors.initials: NK
authors.lastname: Saxena
mesh_terms: D000082:Acetaminophen / Q000008:administration & dosage / Q000037:antagonists & inhibitors* / Q000506:poisoning; D018712:Analgesics, Non-Narcotic / Q000008:administration & dosage / Q000737:chemistry / Q000506:poisoning*; D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D003433:Crosses, Genetic; D005978:Glutathione / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D006470:Hemorrhage / Q000209:etiology / Q000517:prevention & control; D018836:Inflammation Mediators / Q000819:agonists / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism; D007274:Injections, Intraperitoneal; D015227:Lipid Peroxidation / Q000187:drug effects; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009336:Necrosis / Q000209:etiology / Q000517:prevention & control; D018384:Oxidative Stress / Q000187:drug effects*; D011897:Random Allocation; D013047:Specific Pathogen-Free Organisms; D054358:Withanolides / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use*
chemical_list: D018712:Analgesics, Non-Narcotic; D000975:Antioxidants; D018836:Inflammation Mediators; D054358:Withanolides; D000082:Acetaminophen; D005978:Glutathione; C009684:withaferin A
keywords: Acetaminophen; Gclc; Necrosis; Oxidative stress; Withaferin-A