Membrane cholesterol but not putative receptors mediates anandamide-induced hepatocyte apoptosis.
The endogenous cannabinoid anandamide, a lipid mediator, induces various physiologic events such as vascular relaxation, inhibition of gap-junctions formation, tumor proliferation, neurologic analgesia, and apoptosis. Although increased concentration of anandamide in plasma has been implicated in pathophysiologic states including endotoxin-induced hypotension, the effects of anandamide on hepatocytes still remain unclear. In this study, we present evidence that plasma anandamide concentration is highly increased in severe hepatitis and cirrhosis patients. In addition, concentrations of anandamide within the pathophysiologic range potently induced apoptosis of hepatoma cell line (Hep G2) and primary hepatocytes, suggesting a possible link between increased anandamide level and hepatocyte damage. Anandamide-induced cell death was preceded by G0/G1 cell-cycle arrest, activation of proapoptotic signaling (i.e., p38 MAPK and JNK), and inhibition of antiapoptotic signaling (i.e., PKB/Akt) pathways. Moreover, anandamide increased susceptibility to oxidative stress-induced hepatocyte damage. In this context, methyl-beta-cyclodextrin (MCD), a membrane cholesterol depletor, or mevastatin, an HMG-CoA reductase inhibitor, or N-acetyl cysteine, an antioxidant, potently inhibited the anandamide-induced proapoptotic events and cell death, whereas putative cannabinoid receptor antagonists did not exhibit an inhibitory effect on anandamide-induced cell death. Furthermore, binding assay using polymyxin beads revealed that anandamide could interact with cholesterol. In conclusion, our data suggest that cholesterol present in the cell membrane determines the fate of hepatocytes exposed to anandamide, possibly functioning as an anandamide receptor.
Department of Laboratory and Molecular Medicine, Faculty of Medicine, Kagoshima University, Kagoshima, Japan.
Kamal Krishna
KK
Biswas
Krishna Pada
KP
Sarker
Kazuhiro
K
Abeyama
Ko-ichi
K
Kawahara
Satoshi
S
Iino
Yasuharu
Y
Otsubo
Kazuhiko
K
Saigo
Hiroyuki
H
Izumi
Teruto
T
Hashiguchi
Munekazu
M
Yamakuchi
Kazuyo
K
Yamaji
Ryujin
R
Endo
Kazuyuki
K
Suzuki
Hitoshi
H
Imaizumi
Ikuro
I
Maruyama
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001095:Arachidonic Acids / Q000097:blood / Q000502:physiology*; D063385:Cannabinoid Receptor Modulators / Q000097:blood / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000502:physiology*; D063388:Endocannabinoids; D005260:Female; D006505:Hepatitis / Q000097:blood; D022781:Hepatocytes / Q000502:physiology*; D006801:Humans; D018384:Oxidative Stress / Q000502:physiology; D053284:Polyunsaturated Alkamides; D051381:Rats; D017208:Rats, Wistar
D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; D002784:Cholesterol; C078814:anandamide