title: Carbon tetrachloride-induced hepatic injury through formation of oxidized diacylglycerol and activation of the PKC/NF-κB pathway.
abstract: Protein kinase C (PKC) participates in signal transduction, and its overactivation is involved in various types of cell injury. PKC depends on diacylglycerol (DAG) for its activation in vivo We have previously reported that DAG peroxides (DAG-O(O)H) activate PKC in vitro more strongly than unoxidized DAG, suggesting that DAG-O(O)H, if generated in vivo under oxidative stress, would act as an aberrant signal transducer. The present study examined whether DAG-O(O)H are formed in carbon tetrachloride (CCl(4))-induced acute rat liver injury in association with activation of the PKC/nuclear factor (NF)-κB pathway. A single subcutaneous injection of CCl(4) resulted in a marked increase in hepatic DAG-O(O)H content. At the molecular level, immunohistochemistry and subcellular fractionation combined with immunoblotting localized PKCα, βI, βII and δ isoforms to cell membranes, while immunoblotting showed phosphorylation of the p65 subunit of NF-κB, and immunoprecipitation using isoform-specific anti-PKC antibodies revealed specific association of PKCα and p65. In addition, expression of tumor necrosis factor α (TNFα) and neutrophil invasion increased in the CCl(4)-treated rats. Furthermore, we demonstrated that Vitamin E, one of the most important natural antioxidants that suppresses peroxidation of membrane lipids, significantly inhibited the CCl(4)-induced increase in hepatic DAG-O(O)H content and TNFα expression as well as phosphorylation of PKCα and p65. These data demonstrate for the first time that DAG-O(O)H are generated in the process of CCl(4)-induced liver injury, resulting in activation of the PKC/NF-κB pathway and TNFα-mediated aggravation of liver injury.
authors.affiliation: Department of Pathology, Tokai University School of Medicine, Isehara, Kanagawa, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Toriumi
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Horikoshi
authors.forename: R
authors.initials: R
authors.lastname: Yoshiyuki Osamura
authors.forename: Yorihiro
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Naoya
authors.initials: N
authors.lastname: Nakamura
authors.forename: Susumu
authors.initials: S
authors.lastname: Takekoshi
mesh_terms: D055371:Acute Lung Injury / Q000139:chemically induced* / Q000378:metabolism; D000818:Animals; D002251:Carbon Tetrachloride / Q000633:toxicity*; D002458:Cell Fractionation; D004075:Diglycerides / Q000378:metabolism*; D015151:Immunoblotting; D007150:Immunohistochemistry; D016328:NF-kappa B / Q000378:metabolism*; D010545:Peroxides / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D004075:Diglycerides; D016328:NF-kappa B; D010545:Peroxides; D020033:Protein Isoforms; D002251:Carbon Tetrachloride; D011493:Protein Kinase C