降钙素基因相关肽对大鼠血管平滑肌细胞CDK2和Cyclin E的影响

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (3): 249-253.

降钙素基因相关肽对大鼠血管平滑肌细胞CDK2和Cyclin E的影响

邓水秀¹, 曾泗宇², 任俊芳², 郑元斌², 廖端芳², 秦旭平²

¹湖南环境生物职业技术学院药学系, 衡阳 421005,湖南;
²南华大学药物药理研究所, 衡阳 421001,湖南

收稿日期:2009-12-21 修回日期:2010-03-05 出版日期:2011-03-26 发布日期:2011-05-18
通讯作者: 秦旭平,男,教授,硕士生导师,研究方向:心血管重构的药物防治。Tel: 0734-8281587 E-mail: qinxp333@hotmail.com
作者简介:邓水秀,女,硕士,助教,研究方向:心血管药理学。Tel: 0734-8198721 E-mail: tina20040152@yahoo.com.cn廖端芳,男,教授,博士生导师,研究方向:心血管药理学。
基金资助:
国家自然科学基金(30572192)和湖南省自然科学基金(05JJ30042)资助课题

Effect of CGRP on the CDK2 and Cyclin E of rat vascular smooth muscle cell

DENG Shui-xiu¹, ZENG Si-yu², REN Jun-fang², ZHENG Yuan-bin², LIAO Duan-fang², QIN Xu-ping²

¹Department of Pharmacy, Hunan Environment-Biological Polytechnic College, Hengyang　421005, Hunan, China;
²Institute of Pharmacy and Pharmacology, University of South China, Hengyang　421001, Hunan, China

Received:2009-12-21 Revised:2010-03-05 Online:2011-03-26 Published:2011-05-18

摘要/Abstract

摘要： 目的: 研究降钙素基因相关肽(CGRP)对大鼠血管平滑肌细胞中细胞周期蛋白依赖性激酶2(cyclin-dependent kinase 2,CDK2)和细胞周期蛋白E(Cyclin E)的影响,为阐明CGRP抑制血管平滑肌细胞增殖的细胞周期机制提供实验依据。方法: 培养大鼠胸主动脉血管平滑肌细胞,分别用CGRP或/和10%FBS处理细胞。噻唑蓝比色法(MTT)观察CGRP对10%FBS诱导大鼠血管平滑肌细胞增殖的影响;流式细胞术分析细胞周期;Western blot检测CDK2和Cyclin E的表达。结果: CGRP能抑制10%FBS诱导的血管平滑肌细胞增殖,升高G₀/G₁期细胞百分比,降低S+G₂+M期细胞百分比,抑制细胞内CDK2和Cyclin E表达。结论: CGRP能通过阻滞细胞周期由G₀/G₁期进入S期而抑制血管平滑肌细胞增殖,其作用机制可能与降低CDK2和Cyclin E表达有关。

Abstract: AIM: To study the effect of calcitonin gene-related peptide (CGRP) on cyclin-dependent kinase 2(CDK2) and Cyclin E of vascular smooth muscle cell (VSMC), and to elute the mechanism of inhibitory effect of CGRP on proliferation of VSMC in cell cycles. METHODS: VSMC were prepared from thoracic aortas of male Sprague-Dawley rat by explanting method. The cell viability and cell cycle were determined by MTT and Flow Cytometry, respectively. Western Blot was used to observe expressions of CDK2 and Cyclin E of VSMC. RESULTS: Pretreatment of VSMC with CGRP significantly inhibited cells viability, decreased the proportion of S phase and increased ratio of G₀/G₁ that were induced by 10% FBS, simultaneously, CGRP down-regulated the expressions of CDK2 and Cyclin E when cultured cells with 10% FBS at 12, 24 and 48 hours.CONCLUSION: CGRP could inhibit the cell cycle of VSMC G₀/G₁ phase to S phase transition, by which the mechanism maybe involved in the decrease expressions of CDK2 and Cyclin E.

Key words: CGRP, Vascular smooth muscle cell, Cell cycle, CDK2, Cyclin E

中图分类号:

R965.2

邓水秀, 曾泗宇, 任俊芳, 郑元斌, 廖端芳, 秦旭平. 降钙素基因相关肽对大鼠血管平滑肌细胞CDK2和Cyclin E的影响[J]. 中国临床药理学与治疗学, 2011, 16(3): 249-253.

DENG Shui-xiu, ZENG Si-yu, REN Jun-fang, ZHENG Yuan-bin, LIAO Duan-fang, QIN Xu-ping. Effect of CGRP on the CDK2 and Cyclin E of rat vascular smooth muscle cell[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(3): 249-253.

参考文献

[1] Kwasaki H, Takasaki K, Saito A, et al. Calcitonin gene-related peptide acts as a novel vasodilator neurotransmitter in mesenteric resistance vessels of the rat[J]. Nature, 1988, 335(6186):164-167.
[2] Qin XP, Ye F, Liao DF, et al. Involvement of calcitonin gene-related peptide in the depressor effects of losartan and perindopril in rats[J]. Eur J Pharmacol, 2003, 464 (1):63-67.
[3] Qin XP, Chen Y, Qin YF, et al. Different responses of calcitonin gene-related peptide between losartan and prazosin in early phase of renovascular hypertension[J]. 中国临床药理学与治疗学, 2009,14(1):42-47.
[4] Li Y, Fiscus RR, Wu J, et al. The antiproliferative effects of calcitonin gene-related peptide in different passages of cultured vascular smooth muscle cells[J]. Neuropeptides, 1997, 31(5):503-509.
[5] Qin XP, Ye F, Hu CP, et al. Effect of CGRP on proliferation induced by angiotensinⅡin vascular smooth muscle cells[J]. Eur J Pharmaco, 2004, 488:45-49.
[6] 武旭东, 徐成斌, 陈红,等. 降钙素基因相关肽对内皮素致血管平滑肌细胞增殖的影响[J]. 北京医科大学学报,1997, 29(1):91-94.
[7] 李晓艳,黄从新,孙有刚,等.降钙素基因相关肽对人血管平滑肌细胞增殖的抑制作用[J]. 中国病理生理杂志, 2000, 16(1):24-26.
[8] Wang X, Wang W, Li Y, et al. Mechanism of SNAP potentiating antiproliferative effect of calcitonin gene-related peptide in cultured vascular smooth muscle cells[J]. J Mol Cell Cardiol, 1999, 31 (9):1599-1606.
[9] Chattergoon NN, D Souza FM, Deng W, et al. Antiproliferative effects of calcitonin gene-related peptide in aortic and pulmonary artery smooth muscle cells[J]. Am J Physiol Lung Cell Mol Physiol, 2005, 288(1):L202-L211.
[10] Sherr CJ, Roberts JM. CDK inhibitors: positive and negative regulators of G₁-phase progression[J]. Genes Dev, 1999, 13 (12):1501-1512.
[11] Dulic V, Lees E, Reed SI. Association of humane cyclin E with a periodic G₁-S phase protein kinase[J]. Science, 1992, 257 (5078):1958-1961.

[1]	叶凯丽, 郑雯, 叶啟发, 杨岚. CDK1参与肝细胞癌的发展机制及其抑制剂应用价值[J]. 中国临床药理学与治疗学, 2021, 26(9): 1086-1094.
[2]	聂娟，曹玉梅，杨冬梅，孙四玉，王巧琼，谢雪姣，庹勤慧. 夏枯草水提取物对血管平滑肌细胞增殖的影响及其机制研究[J]. 中国临床药理学与治疗学, 2019, 24(9): 977-982.
[3]	吴丽梅，程丽艳，郑晓亮，王孝举. 靶向抑制CDK4/6的肿瘤治疗基础研究与临床应用进展[J]. 中国临床药理学与治疗学, 2019, 24(9): 1065-1069.
[4]	刘亚东，卢小伟，刘志刚. 促红细胞生成素对急性脊髓损伤患者血清降钙素基因相关肽的影响[J]. 中国临床药理学与治疗学, 2019, 24(8): 933-937.
[5]	全海燕，刘玉环，杨丽，阳芳，秦旭平. CGRP通过激活cAMP/PPARγ/eNOS途径改善血管内皮细胞胰岛素抵抗[J]. 中国临床药理学与治疗学, 2019, 24(6): 615-622.
[6]	邱飞, 高丹, 杨冬梅, 钟小妹, 唐莎, 尹辉明, 张在其. 姜黄素对血小板源性生长因子诱导的血管平滑肌细胞增殖及PTEN表达的影响[J]. 中国临床药理学与治疗学, 2019, 24(12): 1358-1363.
[7]	秦博宇，王刚，齐晓光，王雅捷，孙琼，孙胜杰 . 顺铂与唑来膦酸序贯应用抑制肺癌细胞增殖的研究[J]. 中国临床药理学与治疗学, 2018, 23(9): 1027-1030.
[8]	阎卉芳，徐昊，彭熙炜，朱嘉欢，邓常清. 黄芪和当归配伍对大鼠血管内膜增生血管平滑肌细胞增殖的影响[J]. 中国临床药理学与治疗学, 2018, 23(4): 361-369.
[9]	颜为红，邹坤，贺海波，张永峰，李小妹，李小琴，杨小姣，汪鋆植，邓张双. 吉祥草中甾体皂苷RCE-4对人宫颈癌Caski细胞Ras/Erk和p16/cyclinD1/CDK4通路的影响[J]. 中国临床药理学与治疗学, 2018, 23(3): 247-254.
[10]	李献超，马晓东，郑青秀，刘刚. 桑白利湿汤治疗感染后咳嗽的疗效和对诱导痰中P物质、降钙素基因相关肽水平的影响[J]. 中国临床药理学与治疗学, 2018, 23(11): 1282-1286.
[11]	张晓一，于潇华，刘玉环，杨丽，王珍，阳芳，秦旭平，曾泗宇. ERK1/2和p38MAPK在介导CGRP和Ang II诱导血管平滑肌细胞Nox1表达中的作用[J]. 中国临床药理学与治疗学, 2017, 22(1): 13-19.
[12]	杨冬梅，聂娟，孙四玉，邱飞，刘杨，王炜，熊国祚，廖端芳，庹勤慧. 竹节参皂苷IVa甲酯对血管紧张素II诱导的血管平滑肌细胞增殖影响及机制研究[J]. 中国临床药理学与治疗学, 2016, 21(8): 873-877.
[13]	郑彬，孙峰. 黄药子醇提物对人胃癌细胞凋亡及FABP-5表达的影响[J]. 中国临床药理学与治疗学, 2016, 21(3): 252-258.
[14]	何艳鲁卫华秦雪梅金孝岠. 脓毒症大鼠肠道瞬时感受器电位香草酸受体1及降钙素基因相关肽的表达及作用[J]. 中国临床药理学与治疗学, 2016, 21(1): 38-41.
[15]	张学非, 朱萱萱, 刘沈林, 王琼, 严士海, 王海丹, 张迪, 符蕊. 胃瘤安不同组方对人胃癌SGC-7901增殖、细胞周期和凋亡的影响[J]. 中国临床药理学与治疗学, 2015, 20(1): 31-37.