Wiecha J, Schläger B, Voisard R, Hannekum A, Mattfeldt T, Hombach V
Department of Cardiology, University Hospital of Giessen, Germany.
Basic Res Cardiol. 1997 Aug;92(4):233-9. doi: 10.1007/BF00788518.
The behavior of Ca(2+)-activated K+ channels of large conductance (BKCa) in smooth muscle cells, which were obtained from atherosclerotic plaque material (SMCP) and from media segments (SMCM) of human coronary arteries, were compared using the patch-clamp technique. Voltage-clamp protocols in cell-attached patches revealed the characteristic voltage-dependent activation of BKCa in both cell groups. Single-channel conduction as 216.4 +/- 16.7 pS (n = 6) in SMCP and 199.9 +/- 6.7 pS (n = 6) in SMCM in symmetrical 140 mM K+ solutions. Using outside-out patches, external perfusion with 500 microM tetraethylammonium ions caused a typical "flickery block" of the unitary current. The selective BKCa channel inhibitor iberiotoxin (50 nM) effectively blocked BKCa, channel activity. Comparing BKCa open-state probabilities (P0) at +80 mV in cell-attached patches, a highly significant difference between SMCP (P0 = 0.1438 +/- 0.1301; n = 15) and SMCM (P0 = 0.0093 +/- 0.0044; n = 15; Kruskal-Wallis test, p < 0.001) was found. In contrast to this finding, there was no significant difference in the open-state probability of BKCa, between SMCP (P0 = 0.542 +/- 0.0237; n = 9) and SMCM (P0 = 0.0472 +/- 0.0218; n = 10; p = n.s.) using inside-out patches. The results show an interesting difference in the behavior of large conductance Ca(2+)-activated K+ channel in SMCP compared to SMCM with a significantly higher channel activity in human smooth muscle cells obtained from coronary atherosclerotic plaque material. This finding may indicate an important functional role of BKCa channels in the development of atherosclerosis.
采用膜片钳技术比较了从人类冠状动脉粥样硬化斑块组织(SMCP)和中膜段(SMCM)获取的平滑肌细胞中,大电导钙激活钾通道(BKCa)的行为。细胞贴附式膜片钳的电压钳实验方案揭示了两组细胞中BKCa通道典型的电压依赖性激活。在对称的140 mM K+溶液中,SMCP的单通道传导率为216.4±16.7 pS(n = 6),SMCM为199.9±6.7 pS(n = 6)。使用外向膜片,用500 μM四乙铵离子进行外部灌流可引起单位电流典型的“闪烁阻断”。选择性BKCa通道抑制剂iberiotoxin(50 nM)可有效阻断BKCa通道活性。比较细胞贴附式膜片中+80 mV时的BKCa通道开放概率(P0),发现SMCP(P0 = 0.1438±0.1301;n = 15)和SMCM(P0 = 0.0093±0.0044;n = 15;Kruskal-Wallis检验,p < 0.001)之间存在高度显著差异。与该结果相反,使用内向膜片时,SMCP(P0 = 0.542±0.0237;n = 9)和SMCM(P0 = 0.0472±0.0218;n = 10;p = 无显著性差异)的BKCa通道开放概率无显著差异。结果显示,与SMCM相比,SMCP中大电导钙激活钾通道的行为存在有趣的差异,从冠状动脉粥样硬化斑块组织获取的人类平滑肌细胞中通道活性显著更高。这一发现可能表明BKCa通道在动脉粥样硬化发展中具有重要的功能作用。