Carbon monoxide stimulates the Ca2(+)-activated big conductance k channels in cultured human endothelial cells.
We used the whole-cell patch-clamp technique to study K channels in the human umbilical vein endothelial cells and identified a 201 pS K channel, which was blocked by tetraethylammonium and iberiotoxin but not by TRAM34 and apamin. This suggests that the Ca(2+)-activated big-conductance K channel (BK) is expressed in endothelial cells. Application of carbon monoxide (CO) or tricarbonylchloro(glycinato)ruthenium(II), a water soluble CO donor, stimulated the BK channels. Moreover, application of hemin, a substrate of heme oxygenase, mimicked the effect of CO and increased the BK channel activity. The stimulatory effect of hemin was significantly diminished by tin mesoporphyrin, an inhibitor of heme oxygenase. To determine whether the stimulatory effect of CO on the BK channel was mediated by NO and the cGMP-dependent pathway, we examined the effect of CO on BK channels in cells treated with, N(G)-nitro-l-arginine methyl ester, 1H(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one, an inhibitor of soluble guanylate cyclase, or KT5823, an inhibitor of protein kinase G. Addition of either diethylamine NONOate or sodium nitroprusside significantly increased BK channel activity. Inhibition of endogenous NO synthesis with N(G)-nitro-l-arginine methyl ester, blocking soluble guanylate cyclase or protein kinase G, delayed but did not prevent the CO-induced activation of BK channels. Finally, application of an antioxidant agent, ebselen, had no effect on CO-mediated stimulation of BK channels in human umbilical vein endothelial cells. We conclude that BK channels are expressed in human umbilical vein endothelial cells and that they are activated by both CO and NO. CO activates BK channels directly, as well as via a mechanism involving NO or the cGMP-dependent pathway.
Dong
De-Li
DL
Department of Pharmacology, Harbin Medical University, Harbin, China.
Zhang
Yan
Y
Lin
Dao-Hong
DH
Chen
Jun
J
Patschan
Susann
S
Goligorsky
Michael S
MS
Nasjletti
Alberto
A
Yang
Bao-Feng
BF
Wang
Wen-Hui
WH
D000975:Antioxidants / Q000494:pharmacology; D001393:Azoles / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002248:Carbon Monoxide / Q000494:pharmacology*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D054833:Isoindoles; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D008564:Membrane Potentials / Q000502:physiology; D009569:Nitric Oxide / Q000502:physiology; D016566:Organoselenium Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction; D018408:Patch-Clamp Techniques; D015398:Signal Transduction / Q000502:physiology
D000975:Antioxidants; D001393:Azoles; D004791:Enzyme Inhibitors; D054833:Isoindoles; D051036:Large-Conductance Calcium-Activated Potassium Channels; D016566:Organoselenium Compounds; D009569:Nitric Oxide; C042986:ebselen; D002248:Carbon Monoxide; D006152:Cyclic GMP; D002118:Calcium