The roles in agonist-induced morphological mechanisms in cultured human umbilical vein endothelial cells.

培养的人脐静脉内皮细胞中激动剂诱导的形态机制的作用。

• 批准号: 07807010
• 负责人: HASHIGUCHI Toshio
• 金额: $ 1.34万
• 依托单位: Tokyo Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07807010/
• 关键词: Human Umbilical Vein Endothelial Cells; morphometry; roundness value; histamine; protein kinase C; tyrosine kinase; 培養ヒト臍帯静脈血管内皮細胞(HUVEC); roundness(真円度); 培養ヒト血管内皮細胞(HUVEC); フロテインカイネースC; PMA

Vascular endothelium constitutes the inner lining of vascular wall and acts as a selective varrier to various substrates. It is also known that endothelial cells are targets of inflammatory mediators which cause various functions including an increase in vascular permeability. We investigated the roles of agonist-induced morphological changes by time-lapse analysis in cultured human umbilical vein endothelial cells using histamine as an inflammatory mediator. The mechanisms of intracellular signal transductions in inflammation were investigated by the movement of cytosolic Ca^<2+> concentrations and cytoskeletal events. Histamine induced transient shrinkage of cell periphery which coincided with loosening of cell to cell junctions. This change was abolished by promethazine, an antagonist of H^1 receptor, but not H_2 antagonist. This shrinkage was dependent upon extracellular Ca^<2+>. Histamine induced biphasic increase in [Ca^<2+>] _<in>. The first phase was transient and the second on … More e was sustained. An H_1 antagonist was effective on histamine-induced increase in cytosolic Ca^<2+>. These results suggest that histamine activates H^1 receptors. Then influx of Ca^<2+> promotes breakdown of inosytolphospholipid turnover and activation protein kinase C (PKC) which in turn phosphorylates various cellular proteins including cytoskeleton-associated proteins. Diglycerol, one of the products of phospholipid breakdown and an activator of PKC,was found to mimic morphological effect of histamine. On the other hand PMA,another PKC potentiator, induced biphasic morphological changes which is different from that of histamine. The first phase was shrinkage of cell periphery and the second one was delayd elongation of cells. The facts that only the second phase was inhibited by PKC inhibitor, tyrosine kinase inhibitor and protein synthesis inhibitor strongly suggest multiple signaling pathways are involved in histamine action. In addition to PKC activation another pathways presumably through activation of protein tyrosine kinase might play a crucial role in histamine-induced morphological change. Less

血管内皮细胞构成血管壁的内层，是多种底物的选择性屏障。众所周知，内皮细胞是炎症介质的靶标，炎症介质可引起多种功能，包括增加血管通透性。我们使用组胺作为炎症介质，通过时间推移分析研究了激动剂诱导的人脐静脉内皮细胞形态变化的作用。通过细胞内Ca~(2+)浓度和细胞骨架事件的变化，探讨炎症过程中细胞内信号转导的机制。组胺引起细胞周边的一过性收缩，这与细胞与细胞连接的松动相一致。这种变化可被H^1受体拮抗剂异丙嗪所逆转，但不能被H_2受体拮抗剂异丙嗪所阻断。这种收缩依赖于细胞外的钙离子。组胺引起大鼠心肌细胞内[Ca~(2+)]_t；双相升高。第一阶段是暂时性的，第二阶段是…更多的E是持续的。H_1拮抗剂对组胺引起的胞浆Ca~(2+)和Gt~(2+)升高有明显的抑制作用。这些结果表明组胺激活了H^1受体。然后，钙离子的内流促进肌胞内磷脂周转和激活蛋白激酶C(PKC)的分解，进而使包括细胞骨架相关蛋白在内的各种细胞蛋白磷酸化。二甘油是磷脂降解的产物之一，也是PKC的激活剂，被发现与组胺的形态效应相似。另一方面，另一种PKC增强剂PMA引起与组胺不同的双相形态改变。第一阶段为细胞周边收缩，第二阶段为细胞延缓伸长。PKC抑制剂、酪氨酸激酶抑制剂和蛋白质合成抑制剂仅抑制第二时相，这一事实有力地表明组胺作用涉及多条信号通路。在组胺诱导的细胞形态改变中，除PKC激活外，另一条可能通过蛋白酪氨酸激酶激活的途径可能起着至关重要的作用。较少

项目成果

橋口 美津子等: "培養ヒト臍帯静脈内皮細胞のPKC活性化による形態変化" 生化学. 67(7). 632 (1995)

Mitsuko Hashiguchi 等人：“PKC 激活导致的培养人脐静脉内皮细胞的形态变化”，生物化学 67(7) (1995)。

Hashiguchi M.et al.: "Mechanisms of Histamine-induced Morphological Changes of Cultured Human Umbilical Vein Endotherial cells(HUVEC)." The Japanese Journal of Physiology. 46(S). S10- (1996)

Hashiguchi M.et al.：“组胺诱导培养的人脐静脉内皮细胞 (HUVEC) 形态变化的机制。”

Hashiguchi M.et al.: "Mechanisms of Histamine-induced Morphological Changes of Cultured Human Umbilical Vein Endothelial cells(HUVEC)." The Japanese Journal of Physiology. 46(S). S10 (1996)

Hashiguchi M.et al.：“组胺诱导培养的人脐静脉内皮细胞 (HUVEC) 形态变化的机制。”

Hashiguchi M., Hashiguchi T.et al.: "Mechanisms of Histamine-induced Morphological Changes in Cultured Human Umbilical Vein Endothelial cells(HUVEC)." Jpn.J.Physiol.46(S). S10 (1996)

Hashiguchi M.、Hashiguchi T.et al.：“组胺诱导培养的人脐静脉内皮细胞 (HUVEC) 形态变化的机制”。

橋口 美津子等: "PKC活性化による培養ヒト臍帯静脈内皮細胞(HUVEC)の形態変化の定性的、定量的解析" 生化学. 68(7). 1152 (1996)

Mitsuko Hashiguchi 等人：“PKC 激活引起的培养人脐静脉内皮细胞 (HUVEC) 形态变化的定性和定量分析”生物化学 68(7)。