Novel Therapeutic Strategy for Heart Failure : Molecular Mechanism underlying the Regulation of Ca^<2+> Signaling in the Heart

心力衰竭的新治疗策略：心脏中 Ca^<2> 信号传导调节的分子机制

• 批准号: 13307065
• 负责人: NAGAO Taku
• 金额: $ 34.86万
• 依托单位: National Institute of Health Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13307065/
• 关键词: reactive oxygen species; oxidative stress; G protein; seven transmembrane; Ca^<2+> signaling; C^<2+> channel; cardiac myocyte; cardiac hypertrophy; G-タンパク質; システイン; 興奮収縮連関; カルシウムシグナリング; カルシウムチャネル; リアノジン受容体

1) Treatment with H_2O_2 activates G_I/G_o in rat neonatal ventricular myocytes. We found that H_2O_2 modifies Cys^<287> and Cys^<326> of G_<αi>. Angiotensin II stimulation generated ROS and induced the activation of MAP kinase. The elimination of ROS by the co expression of peroxiredoxn II abolished JNK activation induced by angiotensin II without affecting ERK or p38 MAPK activities. These results indicate that ROS, produced by receptor stimulation, serves as an intracellular mediator linking the receptor stimulation and the activation of MARK (JNK).2) Aiming at elucidating the molecular mechanism underlying the gating modulation of L-type Ca^<2+> channels by Ca^<2+> channel modulators, we searched for the DHP binding sites in L-type Ca^<2+> channel a_<1C> subunit. We identified the two key residues, Phe^<1112> and Ser^<1115> in IIIS5-S6 pore-forming region. The double mutant Ca^<2+> channel (F1112A/S115A) was insensitive to Ca^<2+> channel agonists, and weakly blocked by both Ca^<2+ … More > channel agonists and antagonists. We proposed a novel model for the modulation of Ca^<2+> channel gating by the binding of Ca^<2+> channel modulators at the pore-forming region of Ca^<2+> channel α_<1C> subunit.3) We investigated the physiological role of the privileged cross-communication between L-type Ca^<2+> channels and ryanodine receptors. We found that Ca^<2+> channels function as a sensor to the SR Ca^<2+> content to manipulate APD and the total Ca^<2+> influx through Ca^<2+> channels during APs, via the Ca^<2+>-dependent inactivation of Ca^<2+> channels produced by proximal Ca^<2+>-induced Ca^<2+> release CICR-dependent CDI, to ensure the efficacy of CICR.4) In order to elucidate the physiological role of Na^+-Ca^<2+> exchanger (NCX) in cardiac excitation-contraction coupling, we examined the Ca^<2+> signaling in NCX knockout heterozygous mouse heart. We found that the forward mode NCX activity plays a physiologically important role in the regulation of the SR Ca^<2+> content. Less

1）用H_2O_2治疗大鼠新生儿心肌细胞中的G_I/G_O。我们发现H_2O_2修改了g_ <ai>的cys^<287>和cys^<326>。血管紧张素II刺激产生ROS并诱导MAP激酶的激活。通过过氧氧化物II的CO表达消除ROS，消除了血管紧张素II诱导的JNK激活，而不会影响ERK或p38 MAPK活性。这些结果表明，由受体刺激产生的ROS是连接受体刺激的细胞内介体和标记的激活（JNK）.2）。 A_ <1c>亚基。我们在IIIS5-S6孔形成区域中确定了两个关键残基，Phe^<1112>和Ser^<1115>。双突变体Ca^<2+>通道（F1112a/s115a）对Ca^<2+>通道激动剂不敏感，并且被Ca^<2+…更多>通道激动剂和对手所阻断。我们提出了一个新的模型，用于调节Ca^<2+>通道门控在CA^<2+>通道α_<1c> subunit.3的孔形成区域的Ca^<2+>通道调节剂的结合。3）我们研究了L-type Ca^<2+> Channels and ryananosine and ryananodane and ryananodane and l-type ca^<2+> canne nivers and ca^<1c> subunit.3）。我们发现，Ca^<2+>通道功能充当Sr Ca^<2+>内容的传感器，以操纵APD，并通过Ca^<2+> - 通过Ca^<2+> - 通过Ca^<2+> - 通过Ca^<2+>通道的ca^<2+>通道的ca^<2+>通道的近端Ca^<2+> <2+> <2+> <2+> <2+> <2+> <2+> <2+> <2+>的ca^<2+>通道的ca^<2+> - ca^<2+> pancients的总ca^<2+>通道的影响。 CICR.4）为了阐明Na^+-ca^<2+>交换器（NCX）在心脏兴奋 - 诱导偶联中的身体作用，我们检查了NCX敲除杂合小鼠心脏中的Ca^<2+>信号传导。我们发现，正向模式NCX活动在调节SR Ca^<2+>含量中起物理上重要的作用。较少的

项目成果

Sugimoto Y. et al.: "β_1-selective agonist (-)-1-(3,4-dimethoxyphenetylamino)-3-(3,4-dihydroxy)-2-propanol[(-)-RO363] differentially interacts with key amino acids responsible for β_1-selective binding in resting and active states"J. Pharmacol. Exp. Ther.

Sugimoto Y. 等人：“β_1-选择性激动剂 (-)-1-(3,4-二甲氧基苯乙氨基)-3-(3,4-二羟基)-2-丙醇[(-)-RO363] 与关键药物有不同的相互作用在静息和活动状态下负责 β_1-选择性结合的氨基酸“J. Pharmacol. Exp. Ther.

Isogaya M. et al.: "Enhanced cAMP response of naturally occurring mutant of human β_3-adrenergic receptor"Jp. J. Pharmacol.. 88:(3). 314-318 (2002)

Isogaya M.等人：“人β_3-肾上腺素受体天然突变体的增强的cAMP反应”Jp.J.Pharmacol..88：(3)(2002)。

Yamaguchi, S. et al.: "Key roles of Phe^<1112> and Ser^<1115> in the pore-forming IIIS5-S6 linker of L-type Ca^<2+> channel α_<1C> subunit (Cav1.2) in binding of dihydropyridines and action of Ca^<2+> channel agonists"Mol.Phamracol.. (in press). (2003)

Yamaguchi, S. 等人：“Phe^<1112> 和 Ser^<1115> 在 L 型 Ca^<2+> 通道 α_<1C> 亚基的成孔 IIIS5-S6 连接子中的关键作用 (Cav1 .2)二氢吡啶的结合和Ca 2+ 通道激动剂的作用“Mol.Phamracol..(正在出版)。(2003)

Isogaya. M., Nagao, T., and Kurose, H.: "Enhanced cAMP response of the naturally occurring mutant of human β_3-adrenoceptor"Jpn. J. Pharamcol.. 88. 314-318 (2002)

Isogaya. M.、Nagao, T. 和 Kurose, H.：“人 β_3-肾上腺素受体天然突变体的增强 cAMP 反应”J. Pharamcol.. 88. 314-318 (2002)

Hagiwara M. et al.: "High affinity binding of [^3H]DTZ323 to the diltiazem-binding site of L-type Ca^<2+> channels"Eur. J. Pharmacol.. (in press). (2003)

Hagiwara M.等人：“[ 3 H]DTZ323与L型Ca 2 通道的地尔硫卓结合位点的高亲和力结合”Eur。