核因子-κB在凝血酶促自发性高血压大鼠血管平滑肌细胞增殖中的作用

中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (6): 627-633.

核因子-κB在凝血酶促自发性高血压大鼠血管平滑肌细胞增殖中的作用

胡榕¹, 洪华山¹, 许昌声², 吴可贵²

¹福建医科大学附属协和医院心内科, 福州 350001, 福建;
²福建省高血压研究所, 福州 350005, 福建

收稿日期:2008-04-23 修回日期:2008-06-12 发布日期:2020-10-14
通讯作者: 洪华山,男,博士,教授、主任医师,博士生导师,研究方向:心血管基础和临床研究,心力衰竭等。Tel:0591-83357896-8475 E-mail:honghuashan@hotmail.com
作者简介:胡榕,女,医学博士,副主任医师,主要从事心血管疾病的临床工作。Tel:0591-83357896-8491 E-mail:hurong6666@163.com

Effect of nuclear factor-κB on proliferation of vascular smooth muscle cells promoted by thrombin in spontaneously hypertensive rats

HU Rong¹,HONG Hua-shan¹, XU Chang-sheng², WU Ke-gui²

¹Department of Cardiology, Union Hospital, Fujian Medical University, Fuzhou 350001, Fujian, China;
²Hypertension Research Institution of Fujian, Fuzhou 350005, Fujian, China

Received:2008-04-23 Revised:2008-06-12 Published:2020-10-14

摘要/Abstract

摘要： 目的: 研究凝血酶诱导培养的自发性高血压大鼠(SHR)血管平滑肌细胞(VSMCs)的增殖作用,探讨其引起VSMCs增殖与NF-κB途径的关系及可能机制。方法: 以0.01、0.1、0.2、0.5、1.0、2.0 Um/L的凝血酶与培养的VSMCs共孵育24h和用0.5 Um/L凝血酶与VSMCs分别孵育1、2、6、12、24、48h;采用WST-1代谢活性和3H-TdR掺入率测定VSMCs细胞增殖率;免疫荧光法和蛋白质印迹技术检测基础状态下和0.5 Um/L凝血酶干预的VSMCs中NF-κB的细胞内定位及其核中NF-κB的蛋白含量;以已知的AngⅡ和PDGF对VSMCs的增殖作用为阳性对照,研究NF-κB途径与凝血酶诱导的VSMCs增殖关系,100 μmol/L PDTC(NF-κB特异性阻断剂)预处理后检测凝血酶引起VSMCs的增殖率。结果: 浓度为0.2～2.0 Um/L的凝血酶对VSMCs分别有明显的促增殖作用(P<0.05和P<0.01);但浓度为0.5～2.0 Um/L的凝血酶作用24h对VSMCs的促增殖作用差异无统计学意义(P>0.05)。0.5 Um/L凝血酶促VSMCs的增殖呈双峰样改变,1h和24h分别达峰值。基础状态下VSMCs的NF-κB主要分布于细胞浆,凝血酶干预后VSMCs的NF-κB主要分布于细胞核。PDTC预处理明显抑制凝血酶促VSMCs增殖的作用(P<0.01)。结论: 浓度0.2～0.5 Um/L范围内的凝血酶呈浓度依赖性方式促进培养SHR的VSMCs增殖;而且在时间上呈双峰样改变,提示凝血酶的促增殖作用存在延迟现象。凝血酶能够激活VSMCs的NF-κB,使其从细胞浆转位至细胞核。凝血酶引起VSMCs增殖与NF-κB途径有关联,NF-κB可能是VSMC增殖的细胞内信号转导的共同通路。

关键词: 核因子-κB, 凝血酶, 平滑肌细胞, 增殖

Abstract: AIM: To study the dose-effect and time-effect on proliferation of vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR) promoted by hrombin and to explore the relationship between VSMCs proliferation and NF-κB path as well as their possible mechanism.METHODS: VSMCs were incubated with thrombin at the following concentrations of 0.01, 0.1, 0.2, 0.5, 1.0, 2.0 U/mL for 24 h and with 0.5 U/mL thrombin for 1, 2, 6, 12, 24, 48 h respectively;The proliferation rate of VSMCs was determined with both WST-1 metabolic activity and ³H-TdR incorporation efficiency;The expression of NF-κB in VSMCs was tested with both immumofluorescence and western blot in basic condition and with 0.5 U/mL thrombin intervention.Ang Ⅱ and PDGF on proliferation of VSMCs were used as positive control group.In order to research the relationship of NF-κB path and VSMCs proliferation induced by thrombin, after pretreatment with 100 μmol/L PDTC (NF-κB special inhibitor), VSMCs proliferation induced by thrombin was also investigated.RESULTS: Thrombin concentrations of 0.2-2.0 U/mL significantly promoted VSMCs proliferation respectively(P<0.05 or P<0.01).But VSMCs proliferation was not different in the thrombin at the concentrations of 0.5-2.0 U/mL at 24 h (P>0.05).VSMCs proliferation with 0.5 U/mL thrombin exhibited double hump when compared with the Control Group.The peak values at 1 h and 24 h showed that thrombin promotion VSMCs proliferation was not only rapid and brief but also delayed and retarded.The expression of NF-κB of VSMCs in basic condition was mainly distributed in cytoplasmwhile that of VSMCs interfered by thrombin was in nucleus.The PDTC pre-treatment could obviously inhibit VSMCs proliferation induced by thrombin (P<0.01).CONCLUSION: Thrombin promotes VSMCs proliferation in the manner of dose dependent at the concentrations of 0.2-0.5 U/mL.The proliferation curve induced by thrombin exhibits double hump which suggests that thrombin promotion VSMCs proliferation is delayed. Thrombin may activate NF-κB of VSMCs and make it transfer from cytoplasm to nucleus.VSMCs proliferation induced by thrombin is connected with NF-κB path.NF-κB may be the common path for the signal conduction inside the VSMCs proliferation.

Key words: NF-κB, thrombin, vascular smooth muscle cell, proliferation

中图分类号:

R965.2

胡榕, 洪华山, 许昌声, 吴可贵. 核因子-κB在凝血酶促自发性高血压大鼠血管平滑肌细胞增殖中的作用[J]. 中国临床药理学与治疗学, 2008, 13(6): 627-633.

HU Rong,HONG Hua-shan, XU Chang-sheng, WU Ke-gui. Effect of nuclear factor-κB on proliferation of vascular smooth muscle cells promoted by thrombin in spontaneously hypertensive rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(6): 627-633.

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