药学学报, 2015, 50(7): 824-829
引用本文:
刘焕龙, 于丁, 朱忠宁, 苏素文, 陈雪彦, 张永健. 间尼索地平通过Rho/ROCK通路抑制5-羟色胺诱导的大鼠肺动脉平滑肌细胞增殖[J]. 药学学报, 2015, 50(7): 824-829.
LIU Huan-long, YU Ding, ZHU Zhong-ning, SU Su-wen, CHEN Xue-yan, ZHANG Yong-jian. m-Nisodipine inhibited 5-HT-induced proliferation of rat PASMCs through Rho/ROCK signal pathway[J]. Acta Pharmaceutica Sinica, 2015, 50(7): 824-829.

间尼索地平通过Rho/ROCK通路抑制5-羟色胺诱导的大鼠肺动脉平滑肌细胞增殖
刘焕龙1, 于丁1, 朱忠宁2, 苏素文2, 陈雪彦2, 张永健2
1. 河北医科大学 第二医院, 河北 石家庄 050000;
2. 河北医科大学 基础医学院, 河北 石家庄 050017
摘要:
探讨Rho/ROCK通路在5-羟色胺 (5-HT) 诱导的大鼠肺动脉平滑肌细胞 (PASMCs) 增殖中的作用以及间尼索地平 (m-Nis) 对此的影响。组织贴块法原代培养大鼠PASMCs; 噻唑蓝 (MTT) 比色法观察1 μmol·L-1 5-HT作用不同时间对大鼠PASMCs增殖的影响以及m-Nis的干预作用; Western blot及RT-PCR等方法检测5-HT作用下大鼠PASMCs增殖中RhoAROCK1 mRNA和p-MYPT1蛋白表达的变化以及m-Nis对此的影响, 深入探讨其作用机制。结果显示: 1 μmol·L-1 5-HT作用12~72 h可明显诱导大鼠PASMCs增殖 (P < 0.05或P < 0.01); m-Nis (1×10-5、1×10-6、1×10-7和1×10-8 mol·L-1) 均可明显抑制5-HT诱导的大鼠PASMCs增殖, 并呈现一定的浓度依赖性 (P < 0.05或P < 0.01); 另外, 不同浓度m-Nis还明显抑制了5-HT诱导的RhoAROCK1 mRNA的表达以及ROCK的活化 (P < 0.05或P < 0.01)。结果表明Rho/ROCK通路在5-HT诱导的大鼠PASMCs增殖中起重要作用, m-Nis明显抑制了5-HT诱导的大鼠PASMCs增殖, 该作用可能与其抑制Rho/ROCK通路有关。
关键词:    间尼索地平      5-羟色胺      肺动脉平滑肌细胞      增殖      Rho激酶     
m-Nisodipine inhibited 5-HT-induced proliferation of rat PASMCs through Rho/ROCK signal pathway
LIU Huan-long1, YU Ding1, ZHU Zhong-ning2, SU Su-wen2, CHEN Xue-yan2, ZHANG Yong-jian2
1. The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China;
2. School of Basic Medical Sciences, Hebei Medical University, Shijiazhuang 050017, China
Abstract:
This paper is to report the exploration of the activation of Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the inhibitory effect of m-Nis on this pathway. PASMCs were cultured with the explant technique. MTT assay was used to explore the proliferation of PASMCs after 5-HT treated for different time and the intervening effect of m-Nis. RT-PCR and Western blot were used respectively to explore the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 in 5-HT-treated PASMCs and intervening effect of m-Nis. The results of MTT assay suggested that 5-HT (1 μmol·L-1) treatment for 12-72 h significantly induced the proliferation of rat PASMCs (P < 0.05 or P < 0.01), which were inhibited by m-Nis (1×10-5, 1×10-6, 1×10-7, 1×10-8 mol·L-1) in dose-dependent manners (P < 0.05 or P < 0.01). Similarly, the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 were also inhibited by m-Nis in different degrees (P < 0.05 or P < 0.01). Thus, the results of this study suggested that Rho/ROCK pathway played an important role in 5-HT-induced proliferation of rat PASMCs, m-Nis inhibited 5-HT-induced proliferation obviously, which may be related to the blockage of Rho/ROCK signal pathway.
Key words:    m-nisoldipine    5-hydrocrotaline    pulmonary artery smooth muscle cell    proliferation    Rho kinase   
收稿日期: 2014-10-16
基金项目: 国家自然科学基金资助项目 (81273600); 河北省自然科学基金资助项目 (H2013206147, C2011206145).
通讯作者: 陈雪彦, 张永健
Email: cxylong@126.com;zhangyjlm@163.com
相关功能
PDF(1852KB) Free
打印本文
0
作者相关文章
刘焕龙  在本刊中的所有文章
于丁  在本刊中的所有文章
朱忠宁  在本刊中的所有文章
苏素文  在本刊中的所有文章
陈雪彦  在本刊中的所有文章
张永健  在本刊中的所有文章

参考文献:
[1] Suzuki YJ, Day RM, Tan CC, et al. Activation of GATA-4 by serotonin in pulmonary artery smooth muscle cells [J]. J Biol Chem, 2003, 278: 17525-17531.
[2] Wang CC, Gurevich I, Draznin B. Insulin affects vascular smooth muscle cell phenotype and migration via distinct signaling pathways [J]. Diabetes, 2003, 52: 2562-2569.
[3] Lee SL, Simon AR, Wang WW, et al. H2O2 signals 5-HT-induced ERK MAP kinase activation and mitogenesis of smooth muscle cells [J]. Am J Physiol Lung Cell Mol Physiol, 2001, 281: L646-L652.
[4] Brown JH, Del Re DP, Sussman MA. The Rac and Rho hall of fame: a decade of hypertrophic signaling hits [J]. Circ Res, 2006, 98: 730-742.
[5] Chen XY, Liu HL, Pan ZH, et al. Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells [J]. Chin J Cell Biol (细胞生物学杂志), 2009, 31: 853-858.
[6] Sakurada S, Takuwa N, Sugimoto N, et al. Ca2+-dependent activation of Rho and Rho kinase in membrane depolarization-induced and receptor stimulation-induced vascular smooth muscle contraction [J]. Circ Res, 2003, 93: 548-556.
[7] Gong MC, Fujihara H, Somlyo AV, et al. Translocation of RhoA associated with Ca2+ sensitization of smooth muscle [J]. J Biol Chem, 1997, 272: 10704-10709.
[8] Chen XY, Zhang W, Miao QF, et al. m-Nisoldipine attenuates monocrotaline-induced pulmonary hypertension by suppressing 5-HT/ERK MAPK pathway [J]. Acta Pharm Sin (药学学报), 2008, 43: 1011-1018.
[9] Mukai K, Matsushima H, Ishii Y, et al. Effects of calcium on lens epithelial cells in rabbits [J]. Nippon Ganka Gakkai Zasshi, 2006, 110: 361-369.
[10] Liu Y, Suzuki YJ, Day RM, et al. Rho kinase-induced nuclear translocation of ERK1/ERK2 in smooth muscle cell mitogenesis caused by serotonin [J]. Circ Res, 2004, 95: 579-586.
[11] Ohtsu H, Mifune M, Frank GD, et al. Signal-crosstalk between Rho/ROCK and c-Jun NH2-terminal kinase mediates migration of vascular smooth muscle cells stimulated by angiotensin II [J]. Arterioscler Thromb Vasc Biol, 2005, 25: 1831-1836.
[12] Chen XY, Dun JN, Miao QF, et al. Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, suppresses 5-hydroxytryptamine-induced pulmonary artery smooth muscle cell proliferation via JNK and ERK1/2 pathway [J]. Pharmacology, 2009, 83: 67-79.
[13] Ito M, Nakano T, Erdodi F, et al. Myosin phosphatase: structure, regulation and function [J]. Mol Cell Biochem, 2004, 259: 197-209.
相关文献:
1.陈雪彦 刘焕龙 潘振华 苗庆峰 张永健.间尼索地平对5-羟色胺诱导的大鼠肺动脉平滑肌细胞增殖中Ca2+/CaM/CaN通路的影响[J]. 药学学报, 2010,45(1): 49-54
2.陈雪彦;张伟;苗庆峰;张永健.间尼索地平对野百合碱诱导的肺动脉高压防治作用及其抗5-羟色胺—细胞外信号调节激酶通路的机制探讨[J]. 药学学报, 2008,43(10): 1011-1018