Lee Keon Il, Kong In Deok, Baik Soon Koo, Kim Hyun Soo, Lee Dong Ki, Kwon Sang Ok, Chun Boe Gwun
Department of Physiology, Wonju College of Medicine, Yonsei University 162 Ilsan-dong, Wonju 220-701, Korea.
Yonsei Med J. 2004 Aug 31;45(4):649-60. doi: 10.3349/ymj.2004.45.4.649.
Hepatic stellate cells (HSCs) are known to play a role in the pathogenesis of the increased intrahepatic vascular resistance found in chronic liver diseases. The aim of this study was to evaluate the K+ and Ca2+ currents in cultured HSCs from rat liver, through the patch-clamp technique. Most cells were positive for desmin immunostain after isolation and in alpha-smooth muscle actin immunostain after 10 - 14 days of culturing. Outward and inward rectifying K+ currents were confirmed. Two different types of K+ currents were distinguished: one with the inward rectifying current and the other without. The outward K+ currents consisted of at least four components: tetraethylammonium (TEA)-sensitive current, 4-aminopyridine (4-AP)-sensitive current, pimozide-sensitive current and three blocker-resistant current. The peaks of the outward K+ currents evoked by a depolarizing pulse were decreased to 32.0 +/- 3.0, 62.8 +/- 3.7 and 32.8 +/- 3.5% by 5 mM TEA, 2 mM 4-AP and 15 micro M pimozide, respectively. Moreover, the combined application of three blockers caused 86.6 +/- 4.8% suppression. The inward currents evoked hyperpolarizing pulses were inwardly rectifying and almost blocked by Ba2+. Elevation of external K+ increased the inward current amplitude and positively shifted its reversal potential. Voltage- dependent Ca2+ currents which were completely abolished by Cd2+ and nimodipine were detected in 14 day cultured HSCs. In this study, the cultured HSCs were found to express outward K+ currents composed of multiple pharmacological components, Ba2+-sensitive inward rectifying K+ current and L-type Ca2+ current.
肝星状细胞(HSCs)在慢性肝病中肝内血管阻力增加的发病机制中发挥作用。本研究旨在通过膜片钳技术评估大鼠肝脏培养的肝星状细胞中的钾离子(K+)和钙离子(Ca2+)电流。分离后大多数细胞结蛋白免疫染色呈阳性,培养10 - 14天后α-平滑肌肌动蛋白免疫染色呈阳性。证实了外向和内向整流钾电流。区分出两种不同类型的钾电流:一种具有内向整流电流,另一种没有。外向钾电流至少由四个成分组成:四乙铵(TEA)敏感电流、4-氨基吡啶(4-AP)敏感电流、匹莫齐特敏感电流和三种阻滞剂抗性电流。去极化脉冲诱发的外向钾电流峰值分别被5 mM TEA、2 mM 4-AP和15 μM匹莫齐特降低至32.0±3.0%、62.8±3.7%和32.8±3.5%。此外,三种阻滞剂联合应用导致86.6±4.8%的抑制。超极化脉冲诱发的内向电流是内向整流的,几乎被Ba2+阻断。细胞外钾离子浓度升高增加了内向电流幅度并使其反转电位正向移动。在培养14天的肝星状细胞中检测到完全被Cd2+和尼莫地平阻断的电压依赖性钙离子电流。在本研究中,发现培养的肝星状细胞表达由多种药理学成分组成的外向钾电流、Ba2+敏感的内向整流钾电流和L型钙离子电流。