缺血再灌注损伤对大鼠肝细胞钙池操纵的钙通道电流的影响及药物拮抗
黄昌州, 裘法祖, 华中科技大学同济医学院附属同济医院普外科 湖北省武汉市 430030
张宗明,清华大学第一附属医院普外科 北京市 100016
黄昌州,男, 1965-09-08生, 湖北省红安县人, 汉族, 华中科技大学同济医学院附属同济医院普外科博士, 主要从事细胞离子通道与肝胆外科疾病的研究.
国家自然科学基金资助项目, No. 30270532
教育部跨世纪优秀人才培养计划基金资助项目, 教技函〔2002〕48号
上海市教委”曙光计划”基金资助项目, No. 02SG20
通讯作者:张宗明, 100016, 北京市朝阳区酒仙桥一街坊6号, 清华大学第一附属医院普外科. zhangzongming@yahoo.com
电话: 010-64372362 传真: 010-64361322
收稿日期: 2004-11-23 接受日期: 2004-12-28
Effects of 2-APB on store-operated Ca2+channel currents of hepatocytes after hepatic ischemia/reperfusion injuryin rats
Chang-Zhou Huang, Zong-Ming Zhang, Fa-Zu Qiu
Chang-Zhou Huang, Fa-Zu Qiu, Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Zong-Ming Zhang, Department of General Surgery, the First Affiliated Hospital of Tsinghua University, Beijing 100016, China
Supported by National Natural Science Foundation of China, No.30224801; the Trans-Century Excellent Talent Development Plan Fund of Ministry of Education of China, Official Letter No.2002-48; and Shuguang Program Project of Shanghai Educational Committee, No.02SG20.
Correspondence to: Zong-Ming Zhang, Department of General Surgery, First Affiliated Hospital of Tsinghua University, Beijing 100016, China. zhangzongming@yahoo.com
Received: 2004-11-23 Accepted: 2004-12-28
Abstract
AIM: To study the effects of hepatic ischemia/reperfusion (I/R) injury on hepatocellular viability and store-operated calcium channel currents (Isoc) in freshly isolated rat hepatocytes, and the effects of (2-aminoethoxydiphenyl borane) 2-APB on Isoc of hepatocytes after hepatic ischemia/reperfusion injury in rats.
METHODS: Rat hepatic ischemia and reperfusion injury model was established. Hepatocellular viabilities were determined by trypan blue exclusion assay. The effects of 2-APB on Isoc was investigated by whole-cell patch plamp technique.
RESULTS: Ischemia/reperfusion injury significantly reduced hepatocellular viability and increased Isoc in hepatocytes. 2-APB(20, 40, 60, 80, 100 mmol/L)induced a concentration-dependent decrease of Isoc with IC50value of 64.6±10.7 mmol/L(n = 8).
CONCLUSION: Ischemia/reperfusion injury can reduce hepatocellularviability, probably through increasing Isoc in hepatocytes. 2-APB has aprotective effect on ischemia/reperfusion-induced liver injury, probablythough inhibiting Isoc.
Key Words: Hepatocytes; Ischemia/reperfusion; Store-operated Ca2+channel currents; 2-aminoethoxydiphenyl borane
Huang CZ, Zhang ZM, Qiu FZ. Effects of 2-APB on store-operated Ca2+channel currents of hepatocytes after hepatic ischemia/reperfusion injury in rats. Shijie Huaren Xiaohua Zazhi 2005;13(6):739-742
摘要
目的:研究缺血再灌注损伤对大鼠肝细胞活性和钙池操纵的钙通道电流(Isoc)的影响及钙通道阻滞剂(2-aminoethoxydiphenyl borane，2-APB)的拮抗作用.
方法:建立大鼠肝缺血再灌注损伤模型，应用全细胞膜片钳技术研究缺血再灌注损伤对大鼠肝细胞Isoc的影响及2-APB的拮抗作用.
结果:缺血再灌注组肝细胞Isoc为-1 058.0±223.3pA(n = 8)，假手术组为664.5±140.4pA(n = 8)，两组比较差异显著(P<0.05).(2)20、40、60、80、100 mmol/L的2-APB分别使Isoc由给药前的-1 056.7±225.1pA下降至给药后的-853.7±225.1pA、-686.3±145.0pA、 -534.4±120.4pA、-382.6±58.0pA 、-272.9±51.1pA，抑制率分别为19.2±6.2%、35.1±7.6%、49.4±9.9%、63.8±15.8%、74.2±16.5%;给药前后的Isoc比较有显著差异(n= 8，P<0.05或P<0.01);2-APB的抑制作用呈浓度依赖性增强，其EC50为64.6±10.7 mmol/L.
结论:缺血再灌注损伤具有增加大鼠肝细胞的钙离子内流的作用，钙离子内流增加可能导致肝细胞钙超载，从而降低肝细胞的活性;2-APB对缺血再灌注损伤大鼠肝细胞的钙离子内流具有抑制作用，因此对缺血再灌注损伤的肝细胞起到保护作用.
关键词:肝细胞; 缺血再灌注; 钙池操纵的钙通道电流; 钙通道阻滞剂
黄昌州, 张宗明, 裘法祖. 缺血再灌注损伤对大鼠肝细胞钙池操纵的钙通道电流的影响及药物拮抗. 世界华人消化杂志 2005;13(6):739-742
(PDF) 2-APB对大鼠缺血再灌注损伤的肝细胞Isoc的影响. 2-APB的浓度效应(A). 60、80 mM 的2-APB对Isoc影响的I-V曲线(B)(■对照，●60 mmol/L的2-APB，▲80 mmol/L的2-APB).
3 讨论本实验结果显示缺血再灌注损伤能明显降低肝细胞活性，并进一步增加肝细胞的Isoc.肝缺血再灌注损伤后产生大量氧自由基(reactive oxygen species，ROS)，并且ROS引起一系列肝细胞的病理生理事件，如凋亡、坏死和炎症[13-15]，由此推测ROS可能在SOC激活过程中起着重要的作用，我们的其他实验已证实ROS能增加人肝实质细胞的Isoc(资料未显示).Broad et al[16]发现磷脂酶C和多磷酸肌醇是SOC激活所必须的，新近的资料发现在完好无损的细胞中，具有活性的受体调节的磷脂酶C激活瞬时受体电位通道C3(transientreceptor potential channel，TRPC3)是通过甘油二酯产物的作用来完成的，而与蛋白G、蛋白激酶或三磷酸肌醇无关[17-18]，目前认为该类离子通道的本质是瞬时受体电位通道蛋白家族[19].本研究结果表明大鼠肝缺血再灌注损伤后产生大量的氧自由基，进而与肝细胞膜上的受体结合引起膜磷脂酶C的激活，然后激活细胞膜上的SOC通道，导致肝细胞内钙离子超载，从而引起肝细胞损伤.
我们发现2-APB对缺血再灌注损伤后肝细胞Isoc具有明显的抑制作用，且该抑制作用呈浓度依赖性增强，一些新近的研究也证明2-APB可能是一个钙通道直接阻滞剂而不是三磷酸肌醇受体拮抗剂，并且通过细胞外应用2-APB，钙离子释放激活钙电流(Ca2+-release-activated Ca2+current，Icrac)的活性被迅速地抑制，而细胞内应用则不起作用[20-22];此外，Broad et al[16]证实在缺乏三磷酸肌醇受体的DT40细胞上，2-APB能废除由肌浆内质网ATPase抑制剂(Thapsigagin)引起的容积性钙离子内流，这与2-APB直接作用于SOC本身的观点一致，因此我们可以推测2-APB能够通过抑制Isoc来减少钙离子内流，减轻由缺血再灌注损伤所致的肝细胞损害.
总之，我们可以得出如下结论:缺血再灌注损伤具有增加大鼠肝细胞的钙离子内流的作用，钙离子内流增加可能导致肝细胞钙超载，从而降低肝细胞的活性;2-APB对缺血再灌注损伤大鼠肝细胞的钙离子内流具有抑制作用，因此对缺血再灌注损伤的肝细胞起到保护作用.
4 参考文献1 Vazquez G, Wedel BJ, Aziz O, Trebak M, Putney JW Jr. The mammalian TRPC cation channels.
Biochim Biophys Acta 2004;1742:21-36
2 Penner R, Fleig A. Store-operated calcium entry:a tough nut to CRAC. Sci STKE 2004;2004:pe38
3 Putney JW Jr. Store-operated calcium channels: how do we measure them, and why do we care.
Sci STKE 2004;2004:pe37
4 Gregory RB, Hughes R, Riley AM, Potter BV, Wilcox RA, Barritt GJ. Inositol trisphosphate analogues selective for types I
and II inositol trisphosphate receptors exert differential effects on vasopressin-stimulated Ca2+ inflow and Ca2+
release from intracellular stores in rat hepatocytes. Biochem J 2004;381:519-526
5 Rychkov G, Brereton HM, Harland ML, Barritt GJ. Plasma Membrane Ca2+ release-activated Ca2+channels with a
high selectivity for Ca2+ identified by patch-clamp recording in rat liver cells. Hepatology 2001;33:938-947
6 Maruyama T, Kanaji T, Nakade S, Kanno T, Mikoshiba K. 2-APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable
modulator of ins(1, 4, 5)P3-induce Ca2+release. J Biochem (Tokyo) 1997;122:498-505
7 Iwasaki H, Mori Y, Uchida K, Zhou H, Mikoshiba K. 2-aminoethoxydiphenyl borate(2-APB)inhibits capacitative calcium
entry independently of the function of inositol 1, 4, 5-trisphospate receptors. Receptors Channels 2001;7:429-439
8 Bootman MD, Collins TJ, Mackenzie L, Roderick HL, Berridge MJ, Peppiatt CM. 2-aminoethoxydiphenyl borate(2-APB)is
a reliable blocker of store-operated calcium entry but an inconsistent inhibitor of ins P3-induce Ca2+release.
FASEB J 2002;16:1145-1150
9 Braun FJ, Aziz O, Putney JW Jr. 2-aminoethoxydiphenyl borane activates a novel calcium-permeable cation channel.
Mol Pharmacol 2003;63:1304-1311
10 Colletti LM, Remick DG, Burtch GD, Kunkel SL, Strieter RM, Campbell DA Jr. Role of tumor necrosis factor-alpha in
the pathophysiologic alterations after hepatic ischemia/reperfusion injury in the rat. J Clin Invest 1990;85:1936-1943
11 Riccalton-Banks L, Bhandari R, Fry I, Shakeff KM. A simple method for the simultaneous isolation of stellate cells
and hepatocytes from rat liver tissue. Mol Cell Biochem 2003;248:97-102
12 Barros LF, Stutzin A, Calixto A, Catalan M, Castro J, Hetz C,Hermosilla T. Nonselective Cation Channel as effectors of
free radical-induced rat liver cell necrosis. Hepatology 2001;33:114-122
13 Fan C, Zwacka RM, Engelhardt JF. Therapeutic approaches for ischemia/reperfusion injury in the liver. J Mol Med
1999;77:577-592
14 Pardini RS. Toxicity of Oxygen from naturally occurring redox-active pro-oxidants. Arch Insect Biochem Physiol
1995;29:101-118
15 Losser MR, Payen D. Mechanisms of liver damage. Semin Liver Dis 1996;16:357-367
16 Broad LM, Braun FJ, Lievremont JP, Bird GS, Kurosaki T, Putney JW Jr. Role of phospholipase C-inositol 1, 4,5-trisphosphate pathway in calcium release-activated calcium current and capacitatative calcium entry.
J Biol Chem 2001;276:15945-15952
17 Zhang Z, Tang J, Tikunova S, Johnson JD, Chen Z, Qin N, Dietrich A, Stefani E, Birnbaumer L, Zhu MX. Activation Of
Trp3by inositol 1, 4, 5-trisphosphate receptor through displacement of inhibition calmodulin from a common
binding domain. Proc Natl Acad Sci USA 2001;98:3168-3173
18 Trebak M, St J Bird G, McKay RR, Birnbaumer L, Putney JW Jr. Signaling mechanism for receptor-activated canonical
transient receptor potential 3(TRPC3)channels. J Biol Chem 2003;278:16244-16252
19 Clapham DE, Runnels LW, Strubing C. The TRP ion channel family. Natu Rev Neurosci 2001;2:387-396
20 Ma HT, Patterson RL, Van Rossum DB, Birnbaumer L, Mikoshiba K, Gill DL. Requirement of the inositol trisphosphate
receptor for activation of store-operated Ca2+ channels. Science 2000;287:1647-1651
21 Braun FJ, Broad LM, Armstrong DL, Putney JW Jr. Stable activation of single Ca2+ release-activated Ca2+channels
in divalent cation-free solutions. J Biol Chem 2001;276:1063-1070
22 Kukkonen JP, Lund PE, Akerman KE. 2-aminoethoxy-diphenyl borate reveals heterogeneity in receptor-activated
Ca2+discharge and store-operated Ca2+influx. Cell Calcium 2001;30:117-129
编辑 潘伯荣 审读 张海宁( 黄昌州,张宗明, 裘法祖)
张宗明,清华大学第一附属医院普外科 北京市 100016
黄昌州,男, 1965-09-08生, 湖北省红安县人, 汉族, 华中科技大学同济医学院附属同济医院普外科博士, 主要从事细胞离子通道与肝胆外科疾病的研究.
国家自然科学基金资助项目, No. 30270532
教育部跨世纪优秀人才培养计划基金资助项目, 教技函〔2002〕48号
上海市教委”曙光计划”基金资助项目, No. 02SG20
通讯作者:张宗明, 100016, 北京市朝阳区酒仙桥一街坊6号, 清华大学第一附属医院普外科. zhangzongming@yahoo.com
电话: 010-64372362 传真: 010-64361322
收稿日期: 2004-11-23 接受日期: 2004-12-28
Effects of 2-APB on store-operated Ca2+channel currents of hepatocytes after hepatic ischemia/reperfusion injuryin rats
Chang-Zhou Huang, Zong-Ming Zhang, Fa-Zu Qiu
Chang-Zhou Huang, Fa-Zu Qiu, Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Zong-Ming Zhang, Department of General Surgery, the First Affiliated Hospital of Tsinghua University, Beijing 100016, China
Supported by National Natural Science Foundation of China, No.30224801; the Trans-Century Excellent Talent Development Plan Fund of Ministry of Education of China, Official Letter No.2002-48; and Shuguang Program Project of Shanghai Educational Committee, No.02SG20.
Correspondence to: Zong-Ming Zhang, Department of General Surgery, First Affiliated Hospital of Tsinghua University, Beijing 100016, China. zhangzongming@yahoo.com
Received: 2004-11-23 Accepted: 2004-12-28
Abstract
AIM: To study the effects of hepatic ischemia/reperfusion (I/R) injury on hepatocellular viability and store-operated calcium channel currents (Isoc) in freshly isolated rat hepatocytes, and the effects of (2-aminoethoxydiphenyl borane) 2-APB on Isoc of hepatocytes after hepatic ischemia/reperfusion injury in rats.
METHODS: Rat hepatic ischemia and reperfusion injury model was established. Hepatocellular viabilities were determined by trypan blue exclusion assay. The effects of 2-APB on Isoc was investigated by whole-cell patch plamp technique.
RESULTS: Ischemia/reperfusion injury significantly reduced hepatocellular viability and increased Isoc in hepatocytes. 2-APB(20, 40, 60, 80, 100 mmol/L)induced a concentration-dependent decrease of Isoc with IC50value of 64.6±10.7 mmol/L(n = 8).
CONCLUSION: Ischemia/reperfusion injury can reduce hepatocellularviability, probably through increasing Isoc in hepatocytes. 2-APB has aprotective effect on ischemia/reperfusion-induced liver injury, probablythough inhibiting Isoc.
Key Words: Hepatocytes; Ischemia/reperfusion; Store-operated Ca2+channel currents; 2-aminoethoxydiphenyl borane
Huang CZ, Zhang ZM, Qiu FZ. Effects of 2-APB on store-operated Ca2+channel currents of hepatocytes after hepatic ischemia/reperfusion injury in rats. Shijie Huaren Xiaohua Zazhi 2005;13(6):739-742
摘要
目的:研究缺血再灌注损伤对大鼠肝细胞活性和钙池操纵的钙通道电流(Isoc)的影响及钙通道阻滞剂(2-aminoethoxydiphenyl borane，2-APB)的拮抗作用.
方法:建立大鼠肝缺血再灌注损伤模型，应用全细胞膜片钳技术研究缺血再灌注损伤对大鼠肝细胞Isoc的影响及2-APB的拮抗作用.
结果:缺血再灌注组肝细胞Isoc为-1 058.0±223.3pA(n = 8)，假手术组为664.5±140.4pA(n = 8)，两组比较差异显著(P<0.05).(2)20、40、60、80、100 mmol/L的2-APB分别使Isoc由给药前的-1 056.7±225.1pA下降至给药后的-853.7±225.1pA、-686.3±145.0pA、 -534.4±120.4pA、-382.6±58.0pA 、-272.9±51.1pA，抑制率分别为19.2±6.2%、35.1±7.6%、49.4±9.9%、63.8±15.8%、74.2±16.5%;给药前后的Isoc比较有显著差异(n= 8，P<0.05或P<0.01);2-APB的抑制作用呈浓度依赖性增强，其EC50为64.6±10.7 mmol/L.
结论:缺血再灌注损伤具有增加大鼠肝细胞的钙离子内流的作用，钙离子内流增加可能导致肝细胞钙超载，从而降低肝细胞的活性;2-APB对缺血再灌注损伤大鼠肝细胞的钙离子内流具有抑制作用，因此对缺血再灌注损伤的肝细胞起到保护作用.
关键词:肝细胞; 缺血再灌注; 钙池操纵的钙通道电流; 钙通道阻滞剂
黄昌州, 张宗明, 裘法祖. 缺血再灌注损伤对大鼠肝细胞钙池操纵的钙通道电流的影响及药物拮抗. 世界华人消化杂志 2005;13(6):739-742
(PDF) 2-APB对大鼠缺血再灌注损伤的肝细胞Isoc的影响. 2-APB的浓度效应(A). 60、80 mM 的2-APB对Isoc影响的I-V曲线(B)(■对照，●60 mmol/L的2-APB，▲80 mmol/L的2-APB).
3 讨论本实验结果显示缺血再灌注损伤能明显降低肝细胞活性，并进一步增加肝细胞的Isoc.肝缺血再灌注损伤后产生大量氧自由基(reactive oxygen species，ROS)，并且ROS引起一系列肝细胞的病理生理事件，如凋亡、坏死和炎症[13-15]，由此推测ROS可能在SOC激活过程中起着重要的作用，我们的其他实验已证实ROS能增加人肝实质细胞的Isoc(资料未显示).Broad et al[16]发现磷脂酶C和多磷酸肌醇是SOC激活所必须的，新近的资料发现在完好无损的细胞中，具有活性的受体调节的磷脂酶C激活瞬时受体电位通道C3(transientreceptor potential channel，TRPC3)是通过甘油二酯产物的作用来完成的，而与蛋白G、蛋白激酶或三磷酸肌醇无关[17-18]，目前认为该类离子通道的本质是瞬时受体电位通道蛋白家族[19].本研究结果表明大鼠肝缺血再灌注损伤后产生大量的氧自由基，进而与肝细胞膜上的受体结合引起膜磷脂酶C的激活，然后激活细胞膜上的SOC通道，导致肝细胞内钙离子超载，从而引起肝细胞损伤.
我们发现2-APB对缺血再灌注损伤后肝细胞Isoc具有明显的抑制作用，且该抑制作用呈浓度依赖性增强，一些新近的研究也证明2-APB可能是一个钙通道直接阻滞剂而不是三磷酸肌醇受体拮抗剂，并且通过细胞外应用2-APB，钙离子释放激活钙电流(Ca2+-release-activated Ca2+current，Icrac)的活性被迅速地抑制，而细胞内应用则不起作用[20-22];此外，Broad et al[16]证实在缺乏三磷酸肌醇受体的DT40细胞上，2-APB能废除由肌浆内质网ATPase抑制剂(Thapsigagin)引起的容积性钙离子内流，这与2-APB直接作用于SOC本身的观点一致，因此我们可以推测2-APB能够通过抑制Isoc来减少钙离子内流，减轻由缺血再灌注损伤所致的肝细胞损害.
总之，我们可以得出如下结论:缺血再灌注损伤具有增加大鼠肝细胞的钙离子内流的作用，钙离子内流增加可能导致肝细胞钙超载，从而降低肝细胞的活性;2-APB对缺血再灌注损伤大鼠肝细胞的钙离子内流具有抑制作用，因此对缺血再灌注损伤的肝细胞起到保护作用.
4 参考文献1 Vazquez G, Wedel BJ, Aziz O, Trebak M, Putney JW Jr. The mammalian TRPC cation channels.
Biochim Biophys Acta 2004;1742:21-36
2 Penner R, Fleig A. Store-operated calcium entry:a tough nut to CRAC. Sci STKE 2004;2004:pe38
3 Putney JW Jr. Store-operated calcium channels: how do we measure them, and why do we care.
Sci STKE 2004;2004:pe37
4 Gregory RB, Hughes R, Riley AM, Potter BV, Wilcox RA, Barritt GJ. Inositol trisphosphate analogues selective for types I
and II inositol trisphosphate receptors exert differential effects on vasopressin-stimulated Ca2+ inflow and Ca2+
release from intracellular stores in rat hepatocytes. Biochem J 2004;381:519-526
5 Rychkov G, Brereton HM, Harland ML, Barritt GJ. Plasma Membrane Ca2+ release-activated Ca2+channels with a
high selectivity for Ca2+ identified by patch-clamp recording in rat liver cells. Hepatology 2001;33:938-947
6 Maruyama T, Kanaji T, Nakade S, Kanno T, Mikoshiba K. 2-APB, 2-aminoethoxydiphenyl borate, a membrane-penetrable
modulator of ins(1, 4, 5)P3-induce Ca2+release. J Biochem (Tokyo) 1997;122:498-505
7 Iwasaki H, Mori Y, Uchida K, Zhou H, Mikoshiba K. 2-aminoethoxydiphenyl borate(2-APB)inhibits capacitative calcium
entry independently of the function of inositol 1, 4, 5-trisphospate receptors. Receptors Channels 2001;7:429-439
8 Bootman MD, Collins TJ, Mackenzie L, Roderick HL, Berridge MJ, Peppiatt CM. 2-aminoethoxydiphenyl borate(2-APB)is
a reliable blocker of store-operated calcium entry but an inconsistent inhibitor of ins P3-induce Ca2+release.
FASEB J 2002;16:1145-1150
9 Braun FJ, Aziz O, Putney JW Jr. 2-aminoethoxydiphenyl borane activates a novel calcium-permeable cation channel.
Mol Pharmacol 2003;63:1304-1311
10 Colletti LM, Remick DG, Burtch GD, Kunkel SL, Strieter RM, Campbell DA Jr. Role of tumor necrosis factor-alpha in
the pathophysiologic alterations after hepatic ischemia/reperfusion injury in the rat. J Clin Invest 1990;85:1936-1943
11 Riccalton-Banks L, Bhandari R, Fry I, Shakeff KM. A simple method for the simultaneous isolation of stellate cells
and hepatocytes from rat liver tissue. Mol Cell Biochem 2003;248:97-102
12 Barros LF, Stutzin A, Calixto A, Catalan M, Castro J, Hetz C,Hermosilla T. Nonselective Cation Channel as effectors of
free radical-induced rat liver cell necrosis. Hepatology 2001;33:114-122
13 Fan C, Zwacka RM, Engelhardt JF. Therapeutic approaches for ischemia/reperfusion injury in the liver. J Mol Med
1999;77:577-592
14 Pardini RS. Toxicity of Oxygen from naturally occurring redox-active pro-oxidants. Arch Insect Biochem Physiol
1995;29:101-118
15 Losser MR, Payen D. Mechanisms of liver damage. Semin Liver Dis 1996;16:357-367
16 Broad LM, Braun FJ, Lievremont JP, Bird GS, Kurosaki T, Putney JW Jr. Role of phospholipase C-inositol 1, 4,5-trisphosphate pathway in calcium release-activated calcium current and capacitatative calcium entry.
J Biol Chem 2001;276:15945-15952
17 Zhang Z, Tang J, Tikunova S, Johnson JD, Chen Z, Qin N, Dietrich A, Stefani E, Birnbaumer L, Zhu MX. Activation Of
Trp3by inositol 1, 4, 5-trisphosphate receptor through displacement of inhibition calmodulin from a common
binding domain. Proc Natl Acad Sci USA 2001;98:3168-3173
18 Trebak M, St J Bird G, McKay RR, Birnbaumer L, Putney JW Jr. Signaling mechanism for receptor-activated canonical
transient receptor potential 3(TRPC3)channels. J Biol Chem 2003;278:16244-16252
19 Clapham DE, Runnels LW, Strubing C. The TRP ion channel family. Natu Rev Neurosci 2001;2:387-396
20 Ma HT, Patterson RL, Van Rossum DB, Birnbaumer L, Mikoshiba K, Gill DL. Requirement of the inositol trisphosphate
receptor for activation of store-operated Ca2+ channels. Science 2000;287:1647-1651
21 Braun FJ, Broad LM, Armstrong DL, Putney JW Jr. Stable activation of single Ca2+ release-activated Ca2+channels
in divalent cation-free solutions. J Biol Chem 2001;276:1063-1070
22 Kukkonen JP, Lund PE, Akerman KE. 2-aminoethoxy-diphenyl borate reveals heterogeneity in receptor-activated
Ca2+discharge and store-operated Ca2+influx. Cell Calcium 2001;30:117-129
编辑 潘伯荣 审读 张海宁( 黄昌州,张宗明, 裘法祖)