Effect of short-term ethanol treatment on voltage-dependent calcium channels in Kupffer cells.
Kupffer cells, the resident hepatic macrophages, are activated by calcium, and several reports indicate that their function (e.g., phagocytosis and cytokine production) is altered by ethanol. We recently found that Kupffer cells contain L-type voltage-dependent Ca2+ channels. The purpose of this study, therefore, was to evaluate the effect of short-term ethanol treatment on voltage-dependent Ca2+ channels in Kupffer cells. Kupffer cells were isolated from rats 2 hr after intragastric administration of ethanol (5 gm/kg intragastrically). Cytosolic free calcium concentration of cultured Kupffer cells was measured with the fluorescent Ca2+ indicator fura-2. In Kupffer cells isolated from control rats, partial replacement of extracellular Na+ by K+ caused an increase in cytosolic free calcium concentration in a concentration-dependent manner (half-maximal effect was observed with 81 mmol/L K+), presumably because of membrane depolarization. Acute ethanol treatment in vivo shifted the concentration-response curve for K+ to the right (half-maximal effect was observed with 94 mmol/L K+) and reduced the maximal elevation of cytosolic free calcium concentration by means of K+. Significantly, the dihydropyridine-type calcium channel agonist BAY K 8644 (1 mumol/L) shifted the concentration-response curve for K+ to the left in control and ethanol-treated groups (half-maximal effect was observed with 61 and 77 mmol/L K+, respectively). Moreover, the dihydropyridine-type calcium channel blocker nitrendipine (10 mumol/L) prevented the increase in cytosolic free calcium concentration in both groups. When extracellular Ca2+ was omitted from the incubation medium, the increases in cytosolic free calcium concentration due to depolarization were prevented completely. However, direct addition of ethanol to the cell cultures was without effect.(ABSTRACT TRUNCATED AT 250 WORDS)
Hijioka
T
T
Department of Pharmacology, University of North Carolina, Chapel Hill 27599-7365.
Goto
M
M
Lemasters
J J
JJ
Thurman
R G
RG
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000187:drug effects* / Q000502:physiology*; D002478:Cells, Cultured; D004594:Electrophysiology; D000431:Ethanol / Q000097:blood / Q000494:pharmacology*; D005110:Extracellular Space / Q000378:metabolism; D005260:Female; D007425:Intracellular Membranes / Q000378:metabolism; D007728:Kupffer Cells / Q000378:metabolism*; D009568:Nitrendipine / Q000494:pharmacology; D009994:Osmolar Concentration; D011188:Potassium / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D015220:Calcium Channels; D000431:Ethanol; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D009568:Nitrendipine; D011188:Potassium; D002118:Calcium