Yuan X J, Goldman W F, Tod M L, Rubin L J, Blaustein M P
Department of Physiology, University of Maryland School of Medicine, Baltimore 21201.
Am J Physiol. 1993 Feb;264(2 Pt 1):L116-23. doi: 10.1152/ajplung.1993.264.2.L116.
To explore possible mechanisms underlying hypoxia-induced pulmonary vasoconstriction, the effect of hypoxia on outward K+ current (Iout) was evaluated in primary cultured rat pulmonary (PA) and mesenteric (MA) arterial smooth muscle cells using the whole cell patch-clamp technique. When the cells were bathed in standard physiological salt solution and the patch pipettes contained Ca(2+)-free media with 10 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), virtually all of the Iout, including both the rapidly inactivating component (Irt) and the steady-state (noninactivating) component (Iss), was mediated by voltage-gated K+ channels. Reduction of O2 tension in the bath solution from 155 Torr to < 74 Torr with sodium dithionite reversibly inhibited both Irt and Iss in PA myocytes, but not in MA myocytes. The hypoxia-sensitive Iss was activated at about -50 mV; thus, some of the channels responsible for this current may be open at the resting membrane potential (-40 +/- 1 mV) of PA cells used in this study. Hypoxia also significantly depolarized PA cells bathed in PSS (1.8 mM Ca2+) from -40.7 +/- 1.3 to -24.0 +/- 2.4 mV, and PA cells bathed in Ca(2+)-free PSS (0.1 mM EGTA) from -38.4 +/- 1.3 to -26.1 +/- 3.9 mV. The hypoxia-induced inhibition of Iout in PA cells was accompanied by an apparent increase in inward Ca2+ current.(ABSTRACT TRUNCATED AT 250 WORDS)
为了探究低氧诱导肺血管收缩的潜在机制,采用全细胞膜片钳技术,评估低氧对原代培养的大鼠肺(PA)和肠系膜(MA)动脉平滑肌细胞外向钾电流(Iout)的影响。当细胞浸浴在标准生理盐溶液中,且膜片钳微管内含有含10 mM乙二醇双(β-氨基乙醚)-N,N,N',N'-四乙酸(EGTA)的无钙培养基时,几乎所有的Iout,包括快速失活成分(Irt)和稳态(非失活)成分(Iss),均由电压门控钾通道介导。用连二亚硫酸钠将浴液中的氧张力从155 Torr降至<74 Torr,可使PA肌细胞中的Irt和Iss可逆性抑制,但MA肌细胞不受影响。对低氧敏感的Iss在约-50 mV时被激活;因此,负责该电流的一些通道可能在本研究中所用PA细胞的静息膜电位(-40±1 mV)时开放。低氧还使浸浴在PSS(1.8 mM Ca2+)中的PA细胞从-40.7±1.3 mV显著去极化至-24.0±2.4 mV,使浸浴在无钙PSS(0.1 mM EGTA)中的PA细胞从-38.4±1.3 mV去极化至-26.1±3.9 mV。PA细胞中低氧诱导的Iout抑制伴随着内向钙电流的明显增加。(摘要截短于250字)