DL0805抑制血管紧张素Ⅱ通过Ca2+依赖和非Ca2+依赖途径引起的大鼠胸主动脉环收缩

doi:10.3969/j.issn.1006-7795.2013.03.015

首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (3): 398-403.doi: 10.3969/j.issn.1006-7795.2013.03.015

DL0805抑制血管紧张素Ⅱ通过Ca²⁺依赖和非Ca²⁺依赖途径引起的大鼠胸主动脉环收缩

李莉, 袁天翊, 郭晶, 方莲花, 杨海光, 张莉, 杜冠华

中国医学科学院&北京协和医学院药物研究所,药物靶点研究和新药筛选北京市重点实验室,北京 100050

收稿日期:2013-04-08 出版日期:2013-06-21 发布日期:2013-06-17
通讯作者: 杜冠华 E-mail:dugh@imm.ac.cn
基金资助:
"重大新药创制"国家科技重大专项资助项目(2009ZX09302－003,2013ZX09103001－008),国家自然科学基金(81102444, 81102492)。

DL0805 inhibits the vasoconstriction induced by angiotensin Ⅱ in a Ca²⁺-dependent and Ca²⁺-independent manner in the rat aorta

LI Li, YUAN Tianyi, GUO Jing, FANG Lianhua, YANG Haiguang, ZHANG Li, DU Guanhua

Beijing Key Laboratory of Drug Target and Screening Research, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China

Received:2013-04-08 Online:2013-06-21 Published:2013-06-17
Supported by:
This study was supported by the Major Scientific and Technological Special Project for "Significant New Drugs Creation" (2009ZX09302－003, 2013ZX09103001－008), the National Natural Science Foundation of China (81102444, 81102492).

摘要/Abstract

摘要：

目的探讨Rho激酶抑制剂DL0805抑制血管紧张素Ⅱ(angiotensin Ⅱ, Ang Ⅱ)收缩大鼠胸主动脉环的机制。方法体外培养大鼠胸主动脉平滑肌细胞(vascular smooth muscle cell, VSMC),在有/无1.8 mmol/L Ca²⁺的环境下采用Fura2/AM荧光探针法测定细胞内游离钙(intracellular calcium, ［Ca²⁺］_i)的变化。应用Western blotting检测大鼠胸主动脉环肌球蛋白磷酸酶靶亚基1(myosin phosphatase target subunit 1, MYPT1)和Akt蛋白磷酸化,以及Rho激酶1(Rho kinase 1, ROCK1)蛋白表达水平。结果 DL0805(1、10 μmol/L)抑制Ang Ⅱ(100 nmol/L)引起的VSMC细胞内钙释放和外钙内流。DL0805 (25、50 μmol/L)抑制 Ang Ⅱ(100 nmol/L)引起的MYPT1和Akt磷酸化水平以及ROCK1蛋白表达水平增高。结论 DL0805能抑制Ang Ⅱ通过Ca²⁺依赖和非Ca²⁺依赖途径引起的大鼠胸主动脉环收缩。

关键词: DL0805, 血管紧张素Ⅱ, 大鼠胸主动脉环, 细胞内游离钙, Rho 激酶, 血管平滑肌细胞

Abstract:

Objective To investigate the mechanisms of DL0805, a Rho kinase inhibitor, in the relaxation on the contraction induced by angiotensin Ⅱ(Ang Ⅱ) in the rat thoracic aorta. Methods Intracellular ［Ca²⁺］ (［Ca²⁺］i) was measured with Fura2/AM in vascular smooth muscle cell(VSMC). Protein level of Rho kinase 1(ROCK1), the phosphorylation levels of myosin phosphatase target subunit 1 (MYPT1) and Akt in the rat aortic rings were detected by Western blotting. Results DL0805 (1, 10 μmol/L) were shown to inhibit both Ang Ⅱ(100 nmol/L)-induced Ca²⁺ release from internal stores and Ca²⁺ influx. DL0805 (25、50 μmol/L) significantly attenuated the increased protein level of ROCK1, the increased phosphorylation levels of MYPT1 and Akt in the aorta stimulated by Ang Ⅱ(100 nmol/L). Conclusions DL0805 inhibits the vasoconstriction induced by Ang Ⅱ in a Ca²⁺-dependent and Ca²⁺-independent manner in the rat thoracic aorta.

Key words: DL0805, angiotensin Ⅱ, rat thoracic aortic rings, intracellular calcium, Rho kinase, vascular smooth muscle cell

中图分类号:

R 966

李莉, 袁天翊, 郭晶, 方莲花, 杨海光, 张莉, 杜冠华. DL0805抑制血管紧张素Ⅱ通过Ca²⁺依赖和非Ca²⁺依赖途径引起的大鼠胸主动脉环收缩[J]. 首都医科大学学报, 2013, 34(3): 398-403.

LI Li, YUAN Tianyi, GUO Jing, FANG Lianhua, YANG Haiguang, ZHANG Li, DU Guanhua. DL0805 inhibits the vasoconstriction induced by angiotensin Ⅱ in a Ca²⁺-dependent and Ca²⁺-independent manner in the rat aorta[J]. Journal of Capital Medical University, 2013, 34(3): 398-403.

参考文献

[1] Nunes K P, Rigsby C S, Webb R C. RhoA/Rho-kinase and vascular diseases: what is the link[J]. Cell Mol Life Sci, 2010, 67(22): 3823-3836.

[2] Michael S K, Surks H K, Wang Y, et al. High blood pressure arising from a defect in vascular function[J]. Proc Natl Acad Sci USA, 2008, 105(18): 6702-6707.

[3] Somlyo A P, Somlyo A V. Signal transduction and regulation in smooth muscle[J]. Nature, 1994, 372(6503): 231-236.

[4] Mizuno Y, Isotani E, Huang J, et al. Myosin light chain kinase activation and calcium sensitization in smooth muscle in vivo[J]. Am J Physiol Cell Physiol, 2008, 295(2): C358-364.

[5] Somlyo A P, Somlyo A V. Ca²⁺ sensitivity of smooth muscle and nonmuscle myosin Ⅱ: modulated by G proteins, kinases, and myosin phosphatase[J]. Physiol Rev, 2003, 83(4): 1325-1358.

[6] Shimokawa H, Takeshita A. Rho-Kinase is an important therapeutic target in cardiovascular medicine[J]. Arterioscler Thromb Vasc Biol,2005, 25(9):1767-1775.

[7] Gong L L, Fang L H, Peng J H,et al.Integration of virtual screening with high-throughput screening for the identification of novel Rho-kinase I inhibitors[J]. J Biotechnol, 2010, 145(3): 295-303.

[8] Gong L, Peng J,Fang L, et al.The vasorelaxant mechanisms of a Rho kinase inhibitor DL0805 in rat thoracic aorta[J]. Molecules, 2012, 17(5): 5935-5944.

[9] 李莉,袁天翊,杨海光,等. ERK1/2及JNK参与DL0805抑制血管紧张素Ⅱ引起的大鼠离体胸主动脉环收缩[J]. 基础医学与临床, 2013, 33(6): 699-703.

[10] Zhu X M, Fang L H, Li Y J, et al.Endothelium-dependent and -independent relaxation induced by pinocembrin in rat aortic rings[J]. Vascul Pharmacol, 2007, 46(3): 160-165.

[11] Evans J F, Lee J H, Raqolia L. Ang-Ⅱ-induced Ca(2+) influx is mediated by the 1/4/5 subgroup of the transient receptor potential proteins in cultured aortic smooth muscle cells from diabetic Goto-Kakizaki rats[J]. Mol Cell Endocrinol, 2009, 302(1): 49-57.

[12] Nguyen Dinh Cat A, Touyz R M. Cell signaling of angiotensin Ⅱ on vascular tone: novel mechanisms[J]. Curr Hypertens Rep, 2011, 13(2): 122-128.

[13] Wirth A. Rho kinase and hypertension[J]. Biochim Biophys Acta, 2010, 1802(12): 1276-1284.

[14] Touyz R M, Schiffrin E L. Signal transduction mechanisms mediating the physiological and pathophysiological actions of angiotensin Ⅱ in vascular smooth muscle cells[J]. Pharmacol Rev, 2000, 52(4): 639-672.

[15] Lee D L, Webb R C, Jin L. Hypertension and RhoA/Rho-kinase signaling in the vasculature: highlights from the recent literature[J]. Hypertension, 2004, 44(6): 796-799.

DL0805抑制血管紧张素Ⅱ通过Ca²⁺依赖和非Ca²⁺依赖途径引起的大鼠胸主动脉环收缩

DL0805 inhibits the vasoconstriction induced by angiotensin Ⅱ in a Ca²⁺-dependent and Ca²⁺-independent manner in the rat aorta

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 14

编辑推荐

Metrics

本文评价

[1]	边经纬, 尹晓辰, 李浩, 张苏丽, 王美丽, 刘慧荣. 血管紧张素Ⅱ1型受体自身抗体阳性雌鼠孕后出现妊娠期高血压样症状[J]. 首都医科大学学报, 2019, 40(2): 257-262.
[2]	陈晨, 孟艳. 血管紧张素Ⅱ对树突状细胞功能的影响[J]. 首都医科大学学报, 2017, 38(5): 709-714.
[3]	王鹏雁;安建多;王池;王学从;武鑫瑞;王学江. 槲芪散及槲寄生提取物对肝癌细胞内游离钙离子浓度的影响[J]. 首都医科大学学报, 2012, 33(3): 291-296.
[4]	刘飞;李艳芳;曹芳芳;张玲姬;白雪源;吕杨. 麝香保心丸对心力衰竭大鼠肾脏血管紧张素Ⅱ受体各亚型表达的影响[J]. 首都医科大学学报, 2012, 33(1): 99-102.
[5]	王英;王梅. 高磷诱导大鼠血管平滑肌细胞钙化及其电镜表现[J]. 首都医科大学学报, 2009, 30(4): 521-524.
[6]	曹晓菁;李艳芳. 大鼠肾脏α₁-肾上腺素受体和血管紧张素Ⅱ受体亚型表达水平随增龄的改变[J]. 首都医科大学学报, 2008, 29(6): 728-731.
[7]	龚浩;王宇;张忠涛;李建设;马雪梅;周延忠. 血管紧张素Ⅱ及其受体拮抗剂对肝星状细胞Ⅰ型胶原合成的影响[J]. 首都医科大学学报, 2007, 28(1): 7-9.
[8]	周楠;沈颖;焦莉平;王辉;孟群. IgA肾病早期肾脏局部RAS的活化与调节[J]. 首都医科大学学报, 2005, 26(5): 549-554.
[9]	华琦;李东宝;邹海强;陈彪;皮林;刘荣坤. 内皮型一氧化氮合酶4b/a基因多态性与缬沙坦降压疗效的关系[J]. 首都医科大学学报, 2004, 25(3): 349-352.
[10]	田泽君;叶丹;汪家瑞. HSV-tk基因对实验性血管成形术后再狭窄的抑制作用[J]. 首都医科大学学报, 1999, 20(2): 103-106.
[11]	叶丹;汪家瑞;王宏燕;白丹. 巯甲丙脯酸对活化白细胞所致血管平滑肌细胞损伤的保护作用[J]. 首都医科大学学报, 1995, 16(1): 45-47.
[12]	温绍君;何士大;卢德宏;汪家瑞;张维君;胡旭东;陈宝田;赵迎新;龚伊红;罗立群. 心包肽类激素免疫组织化学及免疫电镜的研究[J]. 首都医科大学学报, 1993, 14(3): 192-195.
[13]	盛树力;张万江;赵志炜;郭春远;艾厚喜;韩劲勇;耿军;刘喆;周磊;高键;金键. 中枢和外周脑钠素(BNP)调节醛固酮分泌的不同作用[J]. 首都医科大学学报, 1992, 13(1): 40-45.
[14]	于仲元;何士大;徐宝钢;王晓光;张叔诚;汪家瑞. 人血白细胞血管紧张素Ⅱ样物质的分离提取及鉴定[J]. 首都医科大学学报, 1988, 9(2): 114-116.