The assessment of the contractility of the proliferating vascular smooth muscle

增殖血管平滑肌收缩力的评估

• 批准号: 13832006
• 负责人: NISHIMURA Junji
• 金额: $ 2.37万
• 依托单位: Kyushu 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-13832006/
• 关键词: Vascular smooth muscle; Contraction; Virus vector; Collagen gel; RhoA; Cell culture

The purpose of the present study was to investigate the mechanism underlying the contractility of the proliferating airway smooth muscle. We could obtain the following results.1, Wd could construct the recombinant baculo virus vector, which can be infected with a high efficiency to the mammalian cell, by inserting the cytomegalo virus promoter and the cDNA of interest. This virus could successfully transfected to the NIH 3T3 fibroblast, porcine tracheal smooth muscle cells and porcine coronary smooth muscle cells. It took us long time to optimize the condition for the transfection. However, we could overcome this problem by modifying the purification process of the recombinant baculo virus.2, We could also establish the method to measure isometric tension using the cultured smooth muscle cells by reconstituting these cells into tissue-like preparation. The RhoA transfected cells showed 2-3 times greater contractility, compared with the control treatment.3, On the contrary, the knock-down of RhoA expression by RNA interference greatly inhibited the smooth muscle contractility.

本研究的目的是研究增殖气道平滑肌收缩性的机制。我们得到以下结果：1、Wd通过插入巨细胞病毒启动子和目的cDNA，构建了能够高效感染哺乳动物细胞的重组杆状病毒载体。该病毒能成功转染NIH 3T3成纤维细胞、猪气管平滑肌细胞和猪冠状动脉平滑肌细胞。我们花了很长时间来优化转染条件。然而，我们可以通过改进重组杆状病毒的纯化工艺来克服这个问题。2、我们还可以通过将培养的平滑肌细胞重构为组织样制剂来建立测量等长张力的方法。与对照处理相比，转染RhoA的细胞表现出2-3倍的收缩力。3、相反，通过RNA干扰敲低RhoA的表达极大地抑制了平滑肌的收缩力。

项目成果

Nakamura K., Nishimura J., Hirano K., Ibayashi S., Fujishima M., Kanaide H.: "Hydroxyfasudil, an active metabolite of fasudil hydrochloride, relaxes the rabbit basilar artery by disinhibition of myosin light chain phosphatase"J Cereb Blood Flow Metab. 21.

Nakamura K.、Nishimura J.、Hirano K.、Ibayashi S.、Fujishima M.、Kanaide H.：“羟基法舒地尔是盐酸法舒地尔的活性代谢物，通过解除肌球蛋白轻链磷酸酶的抑制作用来放松兔基底动脉”J Cereb Blood

Setoguchi H, Nishimura J, Hirano K et al.: "Leukotriene C enhances the contraction of porcine tracheal smooth muscle through the activation of Y-27632, a rho kinase inhibitor, sensitive pathway"Br J Pharmacol. 132. 111-118 (2001)

Setoguchi H、Nishimura J、Hirano K 等人：“白三烯 C 通过激活 Y-27632（一种 rho 激酶抑制剂）敏感途径增强猪气管平滑肌的收缩”Br J Pharmacol。

Shintani Y., Hirano K., Nakayama T., Nishimura J., Nakano H., Kanaide H.: "Mechanism of trypsin-induced contraction in the rat myometrium: The possible involvement of a novel member of protease-activated receptor"Br. J. Pharmacol.. 133. 1276-1285 (2001)

Shintani Y.、Hirano K.、Nakayama T.、Nishimura J.、Nakano H.、Kanaide H.：“胰蛋白酶诱导大鼠子宫肌层收缩的机制：蛋白酶激活受体新成员的可能参与”Br

Nakayama T., Hirano K., Shintani Y., Nishimura J., Nakatsuka A., Kuga H., Takahashi S., Kanaide H.: "Unproductive cleavage and inactivation of protease-activated receptor-1 by trypsin in vascular endotheial cells"Br. J. Pharmacol. 138. 121-130 (2003)

Nakayama T.、Hirano K.、Shintani Y.、Nishimura J.、Nakatsuka A.、Kuga H.、Takahashi S.、Kanaide H.：“血管内皮细胞中胰蛋白酶对蛋白酶激活受体 1 的非生产性裂解和失活

Nakayana T., Hirano K., Nishimura J., Tkahashi S., Kanaide H.: "Mechanism of trypsin-induced endothelium-dependent vasorelaxation in the porcine coronary artery"Br J Pharmacol. 134. 815-826 (2001)

Nakayana T.、Hirano K.、Nishimura J.、Tkahashi S.、Kanaide H.：“猪冠状动脉中胰蛋白酶诱导的内皮依赖性血管舒张的机制”Br J Pharmacol。