Relationship between lipid mediator receptors and lipid-induced impairment of human endothelial cells

脂质介质受体与脂质诱导的人内皮细胞损伤的关系

• 批准号: 13672293
• 负责人: MURAKI Katsuhiko
• 金额: $ 2.18万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-13672293/
• 关键词: human endothelial cell; lipid mediator; endothelial dysfunction; Ca; cell death; ion channel

2001Ca^<2+> entry pathway activated by sphingosine-1-phosphate (S1P) was examined in human umbilical vein endothelial cells (HUVECs) by measuring intracellular Ca^<2+> concentration ([Ca^<2+>]_i), whole-cell membrane currents and single channel activity. Application of S1P to HUVECs induced a slowly developing increase in [Ca^<2+>_i. When Ca^<2+> was absent in the bathing solution, S1P did not affect [Ca^<2+>]_i Addition of Ca^<2+> to the bathing solution, however, produced a sustained increase in [Ca^<2+>_i in the presence of S1P, suggesting that influx of Ca^<2+> plays an obligatory role in elevation of [Ca^<2+>_i by S1P. Pretreatment of HUVECs with pertussis toxin (PTX) abolished S1P-induced elevation of [Ca^<2+>]_i. When whole-cell membrane currents were recorded under the monitoring of [Ca^<2+>]_i, the application of S1P induced a tiny inward current (I_<S1P>) which was followed by the elevation of [Ca^<2+>]_i. The reversal potential of I_<S1P> revealed that I_<S1P> is a ***-selec … More tive cation (NSC) current. When S1P was included in the pipette solution in the excised inside-out patch clamp configuration, ***gle channel with a conductance of 17 pS was activated, depending on the presence of intracellular GTP. These results suggest that S1P has a novel function to activate a NSC-channel in a GTP-dependent manner via a PTX-sensitive G-protein. The NSC-channel activated by S1P acts as a Ca^<2+> entry pathway in endothelium and may regulate the cell-proliferation. These results have been published in J. Physiol. (537, 431-441, 2001)2002Effects of palmitoylcarnitine (palcar), a lipid that is released from various types of cells under the ischemic conditions, on [Ca^<2+>]_i were examined in HUVECs and compared with those of S1P. Application of palcar elevated [Ca^<2+>]_i in HUVECs and its potency was about 30 times lower than that of S1P. Response to 3 μM palcar in each HUVEC clearly paralleled that to 0.3 μM S1P. In addition, HUVECs that were treated with PTX failed to respond to palcar as werll as to S1P. These results suggest that palcar has a novel action on huvecs as a potential agonist of receptors for S1P. These results have been published in J. Pharmacol. Sci. (in press, 2003) Less

2001通过测量细胞内Ca^<2+>浓度（[Ca^<2+>]_i）、全细胞膜电流和单通道活性，研究了鞘氨醇-1-磷酸（S1P）激活的Ca^<2+>进入人脐静脉内皮细胞（HUVECs）。S1P在HUVECs中的应用诱导了[Ca^<2+ bbb_i的缓慢增加。当洗浴液中不存在Ca^<2+>时，S1P不影响[Ca^<2+>]_i，然而，在有S1P存在的情况下，Ca^<2+>_i持续增加，表明Ca^<2+>的内流在S1P升高[Ca^<2+>_i中起着必要的作用。百日咳毒素（PTX）预处理HUVECs可消除s1p诱导的[Ca^<2+>]_i升高。在[Ca^<2+>]_i监测下记录全细胞膜电流，S1P的应用诱导了一个微小的内向电流（I_<S1P>），随后升高了[Ca^<2+>]_i。I_<S1P>的逆转电位表明I_<S1P>是一个***选择性多激活阳离子（NSC）电流。当移液液中加入S1P时，根据细胞内GTP的存在，具有17 pS电导的单个通道被激活。这些结果表明，S1P具有通过ptx敏感的g蛋白以gtp依赖的方式激活nsc通道的新功能。S1P激活的nsc -通道作为Ca^<2+>进入内皮细胞的通路，可调节细胞增殖。这些研究结果发表在《J. Physiol》杂志上。[537,431 - 441,2001] 2002研究了棕榈酰肉碱（palcar）对HUVECs [Ca^<2+>]_i的影响，并将其与S1P进行了比较。棕榈酰肉碱是一种在缺血条件下由各种类型的细胞释放的脂质。palcar提高了[Ca^<2+>]_i在HUVECs中的作用，其效力比S1P低约30倍。每个HUVEC对3 μM palcar的响应与0.3 μM S1P的响应明显相似。此外，接受PTX治疗的huvec对palcar和S1P都没有反应。这些结果表明，palcar作为S1P受体的潜在激动剂对huves具有新的作用。这些结果发表在《J. Pharmacol》杂志上。科学。(出版，2003年

项目成果

A. Yamada et al.: "Usefulness and limitation of DiBAC_4(3), a voltage-sensitive fluorescent dye, for the measurement of membrane potentials regulated by recombinant large conductance Ca^<2+>-activated K^+ channels in HEK293 cells"Japanese Journal of Pharm

A. Yamada 等人：“DiBAC_4(3)（一种电压敏感荧光染料）用于测量 HEK293 细胞中重组大电导 Ca^<2>-激活 K^通道调节的膜电位的用途和局限性”

K.Muraki, Y.Imaizumi: "A novel function of sphingosine-1-phosphate to activate a non-selective cation channel in human endothelial cells"Journal of Physiology. 573-2. 431-441 (2001)

K.Muraki、Y.Imaizumi：“1-磷酸鞘氨醇激活人内皮细胞非选择性阳离子通道的新功能”生理学杂志。

S. Ohya et al.: "Molecular cloning and expression of the novel splice variants of K^+ channel-interacting protein 2"Biochemical Biophysical Research Communications. 282. 96-102 (2001)

S.Ohya等人：“Kα通道相互作用蛋白2的新型剪接变体的分子克隆和表达”生物化学生物物理研究通讯。

村木克彦: "平滑筋及び内皮細胞におけるCa関連イオンチャネル電流解析とCa動態 (総説)"日本薬理学会雑誌. 121. 143-151 (2003)

Katsuhiko Muraki：“平滑肌和内皮细胞中的 Ca 相关离子通道电流分析和 Ca 动力学（评论）”日本药理学会杂志 121. 143-151 (2003)。

Y.Ohi, et al.: "Local Ca^<2+> transients and distribution of BK channels and ryanodine receptors in smooth muscle cells of guinea pig vas deferens and urinary bladder"Journal of Physiology. 534. 313-326 (2001)

Y.Ohi等人：“豚鼠输精管和膀胱平滑肌细胞中BK通道和兰尼碱受体的局部Ca^2瞬变和分布”生理学杂志。