Acute and chronic effect of alcohol on Ca2+ channels in hepatic stellate cells.
BACKGROUND
Hepatic stellate cells have been reported to play important roles in the regulation of hepatic microcirculation via cell contraction. Increase in intracellular calcium concentration is required to induce cell contraction. We have already reported the existence of L-type voltage-operated Ca2+ channels (VOCC), such as smooth muscle cells. On the other hand, alcohol has been known to disturb hepatic microcirculation. In this study, we evaluated the effect of acute and chronic treatment of alcohol on VOCC in rat hepatic stellate cells.


METHODS
Stellate cells isolated from rats were cultured with or without 100 mM ethanol for up to 14 days. VOCC were detected by the patch clamp technique. Cells cultured for 14 days without ethanol were exposed to ethanol to investigate calcium current during membrane depolarization. alpha-Smooth muscle actin (alpha-SMA) was stained by indirect immunofluorescence.


RESULTS
In the control model, VOCC were recognized in cells cultured for more than 7 days. Detection of VOCC increased from 9% on day 7 to 55% on day 14. On the other hand, VOCC in cells treated chronically with 100 mM ethanol appeared earlier than in the control and the incidences were significantly higher than those of the control accompanied with an early activation of cells. In contrast, simultaneous exposure to ethanol during the membrane depolarization inhibited Ca2+ current.


CONCLUSIONS
The expression of Ca2+ channels in stellate cells were up-regulated by the chronic treatment of alcohol accompanied with the transformation to myofibroblast-like phenotype. However, alcohol itself inhibited Ca2+ current.
Oide
H
H
Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
Itatsu
T
T
Hirose
M
M
Wang
X E
XE
Nishiyama
D
D
Takei
Y
Y
Sato
N
N
D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D015220:Calcium Channels / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D004727:Endothelium / Q000166:cytology; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar
D000199:Actins; D015220:Calcium Channels; D002492:Central Nervous System Depressants; D000431:Ethanol