中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (46): 8603-8608.doi: 10.3969/j.issn.1673-8225.2011.46.013

• 血管组织构建 vascular tissue construction • 上一篇    下一篇

apelin-13下调caveolae可促进血管平滑肌细胞增殖

毛小环1, 2，李兰芳1，高  晶1，杨  莉1，柳  威1，秦旭平1，陈临溪1   

  1. 1南华大学药物药理研究所，湖南省衡阳市421001
    2湖南环境生物职业技术学院，湖南省衡阳市 421005
  • 收稿日期:2011-05-10 修回日期:2011-07-06 出版日期:2011-11-12 发布日期:2011-11-12
  • 通讯作者: 陈临溪，博士，教授，南华大学药物药理研究所，湖南省衡阳市421001 chenlinxi@tom.com
  • 作者简介:毛小环★，男，1980年生，湖南省衡阳市人，汉族，南华大学在读硕士，讲师，主要从事药理学与生物信息学的教学与科研工作。 csumxh@sina.com 并列第一作者：李兰芳☆，女，1978年生，湖南省衡阳市人，汉族，2010年中国协和医科大学毕业，博士，讲师，主要从事药理学与生物信息学的教学与科研工作。 149083488@qq.com
  • 基金资助:

    国家自然科学基金项目(30901577) 和教育部留学回国人员科研启动基金(20091590)。

apelin-13 promotes proliferation of vascular smooth muscle cells by downregulating caveolae expression

Mao Xiao-huan1, 2, Li Lan-fang1, Gao Jing1, Yang Li1, Liu Wei1, Qin Xu-ping1, Chen Lin-xi1   

  1. 1Institute of Pharmacy and Pharmacology, University of South China, Hengyang 421001, Hunan Province, China
    2Hunan Polytechnic College of Environment and Biology, Hengyang 421005, Hunan Province, China
  • Received:2011-05-10 Revised:2011-07-06 Online:2011-11-12 Published:2011-11-12
  • Contact: Chen Lin-xi, Doctor, Professor, Institute of Pharmacy and Pharmacology, University of South China, Hengyang 420001, Hunan Province, China chenlinxi@tom.com
  • About author:Mao Xiao-huan★, Studying for master’s degree, Lecturer, Institute of Pharmacy and Pharmacology, University of South China, Hengyang 420001, Hunan Province, China; Hunan Polytechnic College of Environment and Biology, Hengyang 420001, Hunan Province, China csumxh@sina.com Li Lan-fang☆, Doctor, Lecturer, Institute of Pharmacy and Pharmacology, University of South China, Hengyang 420001, Hunan Province, China 149083488@qq.com Ma Xiao-huan and Li Lan-fang contributed equally to this paper and were considered as co-first authors.
  • Supported by:

    the National Natural Science Foundation of China, No.30901577*; the Scientific Research Foundation for Returned Overseas Chinese Scholars, State Education Ministry, No. 20091590*

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被引次数

1

摘要：

背景：结合课题组以往研究成果，提出caveolae可能参与apelin-13促血管平滑肌细胞增殖的假设。
目的：实验观察细胞膜特殊凹陷结构caveolae参与G蛋白偶联受体APJ的内源性配体apelin-13促进大鼠血管平滑肌细胞增殖的作用。
方法：采用组织贴块法培养大鼠胸主动脉血管平滑肌细胞，用MTT方法观察血管平滑肌细胞增殖，Western Blotting方法观察信号蛋白表达，免疫共沉淀技术检测信号分子复合物的形成。
结果与结论：①caveolae结构破坏剂β-环糊精(5 mmol/L，25 h)可明显增强apelin-13诱导的血管平滑肌细胞增殖。②apelin-13(0，1，2，4，8 µmol/L)刺激血管平滑肌细胞，caveolin-1的表达下调，在1 µmol/L时下调明显。③β-环糊精        (5 mmol/L)破坏caveolae后，可使apelin-13下调caveolin-1表达的作用增强。④对照组(体积分数为0.1%胎牛血清孵育)及处理组(apelin-13刺激)caveolin-1与PI3K及ERK1/2均有复合物形成，在apelin-13刺激的情况caveolin-1-PI3K复合物减少、caveolin-1-ERK1/2复合物减少，即apelin-13可能促进caveolin-1与PI3K及ERK1/2解离。结果提示细胞膜特殊凹陷结构caveolae参与apelin-13促血管平滑肌细胞增殖作用。

关键词: 血管平滑肌细胞, apelin-13, APJ, caveolae, caveolin-1, β-环糊精, PI3K, ERK1/2

Abstract:

BACKGROUND: Previous results have shown that caveolae is likely to participate in apelin-13 promotion of vascular smooth muscle cell proliferation.
OBJECTIVE: APJ is a G-protein coupled receptor, and its ligand is apelin peptide. Previously, we reported PI3K and ERK1/2 signal pathway mediated proliferation of vascular smooth muscle cells (VSMCs) induced by apelin-13. This study is to observe caveolae involved in rat VSMCs proliferation induced by apelin-13 and to determine whether caveolin-1 protein binds to signal molecules PI3K, ERK1/2 to mediate VSMC proliferation induced by apelin-13.
METHODS: VSMCs were prepared from male Sprague-Dawley rat thoracic aorta by the primary-explant method. The effect of β- cyclodextrin on cell proliferation induced by apelin-13 was measured by MTT assay. VSMCs were incubated with apelin-13 and β-cyclodextrin (β-CD), caveolin-1 expression was detected by western blot assay. Formation of high molecular weight polyprotein component including caveolin-1 and PI3K /ERK1/2 was detected by immunoprecipitation.
RESULTS AND CONCLUSION: β-CD (5 mmol/L, 25 hours) significantly enhanced VSMCs proliferation induced by apelin-13   (P< 0.05). Treating VSMCs with apelin-13 (0, 1, 2, 4, 8 µmol/L) downregulated apelin-13-induced expression of caveolin-1 and the effect was distinct at 1µmol/L (P < 0.05). Pretreatment of the cells with 5 mmol/L β-CD could strength the downregulation of apelin-13-induced caveolin-1 expression (P < 0.05). Apelin-13 may induce dissociation of PI3K (ERK1/2) with caveolin-1 in VSMCs compared with the control group. These findings suggest that caveolae is involved in apelin-13-induced VSMCs proliferation.

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