title: Sphingosine 1-phosphate protects rat liver sinusoidal endothelial cells from ethanol-induced apoptosis: Role of intracellular calcium and nitric oxide.
abstract: In alcoholic liver disease, ethanol-induced damage to sinusoidal endothelial cells (SECs) appears to be important in the progression of liver damage. However, little is known about the mechanisms responsible for protection of SECs against ethanol-induced injury. To elucidate the role of sphingosine 1-phosphate (S1P), which is stored in platelets and may be released from them on their activation, we investigated the effect of S1P on rat liver SECs in primary culture. Pretreatment of cells with 1 mumol/L S1P attenuated ethanol-induced apoptosis. Electron microscopy confirmed this protective effect of S1P on damaged SECs in liver tissues after perfusion of ethanol. In the absence of ethanol, S1P increased DNA synthesis as determined via incorporation of bromodeoxyuridine. S1P also ameliorated the decreased DNA synthesis of cells induced by ethanol. Addition of S1P to cells induced an increase in intracellular calcium concentrations and NO production in cells. Western blotting revealed that S1P significantly induced the activation of endothelial NO synthase (eNOS), but not Akt, and that S1P-induced activation of eNOS was blocked by trifluoperazine, a calmodulin inhibitor. Furthermore, N(G)-nitro-L-arginine methyl ester, a NO synthase inhibitor, cancelled the effect of S1P on DNA synthesis, apoptosis, and NO production in vitro as well as the protective effect of S1P on cell damage in situ. In conclusion, the biological effect of S1P is at least partially mediated by Ca(2+)-sensitive eNOS activation and subsequent NO formation; extracellular S1P could contribute to sinusoidal protection and remodeling in alcoholic liver injury.
authors.affiliation: Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Zheng
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Kitamura
authors.forename: Kenichi
authors.initials: K
authors.lastname: Ikejima
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Enomoto
authors.forename: Shunhei
authors.initials: S
authors.lastname: Yamashina
authors.forename: Satoko
authors.initials: S
authors.lastname: Suzuki
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Takei
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Sato
mesh_terms: D000818:Animals; D000891:Anti-Infective Agents, Local; D017209:Apoptosis / Q000187:drug effects*; D001973:Bromodeoxyuridine / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000648:ultrastructure; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000009:adverse effects; D005260:Female; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008246:Lysophospholipids / Q000494:pharmacology*; D008854:Microscopy, Electron; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014268:Trifluoperazine
chemical_list: D000891:Anti-Infective Agents, Local; D002147:Calmodulin; D008246:Lysophospholipids; D014268:Trifluoperazine; C060506:sphingosine 1-phosphate; D000431:Ethanol; D004247:DNA; D052250:Nitric Oxide Synthase Type III; C494920:Akt1 protein, rat; D051057:Proto-Oncogene Proteins c-akt; D001973:Bromodeoxyuridine; D013110:Sphingosine; D002118:Calcium