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内皮素A型受体反义基因减弱内皮细胞条件培养液刺激的血管平滑肌细胞增殖(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2004年第2期
页码:
95-98,102
栏目:
基础研究
出版日期:
2004-03-01

文章信息/Info

Title:
Antisense oligonucleotide of endothelin receptor A in attenuating the proliferation of human saphenous smooth muscle cells induced by endothelial cell conditioned medium
作者:
钱济先1; 宋胜云2; 马保安1; 范清宇1
1.第四军医大学唐都医院全军骨肿瘤研究所暨全军骨科中心,陕西 西安 710038; 2.云南武警总队宝山医院外科,云南 宝山 678000
Author(s):
QIAN Jix-ian1; SONG Sheng yun2; MA Bao-an1; FAN Qing-yu1
1.Department of Orthopedics Oncology Institute and Orthopedics Surgery Center of PLA, Tangdu Hospital Fourth Militar Medical Universit, Shaani Xi'an 710038, 2.Surgery Department, Yunnan General Team Hospital of Armed Police, Baoshan, Yunnan 678000
关键词:
内皮素受体; 反义寡核苷酸; 血管平滑肌细胞; 内皮细胞; 大隐静脉
Keywords:
endothelin receptor;  antisense oligonucleotide;  endothelial cell;  smooth muscle cell;  saphenous vein
分类号:
R540.47
DOI:
-
文献标识码:
A
摘要:
目的:研究内皮素(Endothelin,ET)A型受体(ETA)反义寡核苷酸(Oligonucleotide,OGN)对内皮细胞(Endothelialcells ECs)条件培养液(ECCM)刺激的人大隐静脉(humansaphenousvein,HSV)血管平滑肌细胞(Vascularsmoothmusclecells, VSMCs)增殖的影响。方法:分离、培养HSV的ECs和VSMCs。收集ECCM,分别用正常的和加入DMEM(Dulbecco'sModIfiedEaglemedium)、含ET-1的 DMEM、ECCM和加入Phophoramidon(ET-1转化酶抑制剂)的ECCM培养VSMCs。以放射免疫法测定ECs培养基中ET-1水平;分别采用放射受体结合分析法和四唑盐(MTT)比色实验检测ETA蛋白表达和细胞的增殖。结果:ETA-OGN(15mol/L)以时间依赖的方式抑制ETA表达。12～48 hHSV的ECs培养上清液ET-1水平呈现上升的趋势,48 h达到高峰。ECCM显著刺激VSMCs增殖(P<0.01),ET-1能够模拟 ECCM的促增殖作用。在ECCM中加入 Phosphorimidon,不能使 ECCM的促增殖效应消失(P<0.05)。ETA-OGN(15mol/L)对无血清正常的DMEM培养基培养的VSMCs增殖没有显著影响,但显著抑制ECCM ET-1的促增殖作用(P<0.01vsECCM)。结论: ECs通过分泌包括 ET-1在内的多种因子促进VSMCs增殖,其中 ET-1发挥主要的作用。ETA-OGN抑制ECs的促增殖作用,减弱VSMCs增殖,可能是其防治血管移植术后狭窄的重要机制。
Abstract:
AIM:To investigate the effect of antisense oligonucleotide of endothelin receptor A (ETA-OGN) on the proliferation of vascular smooth muscle cells (VSMCs) from human saphenousvein(HSV) stimulated by endothelial cell conditioned medium(ECCM). METHODS: VSMCs and ECs were isolated from HSV by explant culture. ECCM was harvested and VSMCs were cultured respectively in DMEM (Dulbecco's Midufued Eagle medium), ECCM, ECCM conditioned by adding phosphoramidon (in hibitor of endothelial converting enzyme), and DMEM withET-1. Radioimmunoassay was applied for detecting ET-1 secretion of ECs. The growth and ETA expression of VSMCs was measured by means of MTT method and 125I ET-1 radiobinding assay respectively. RESULTS: ETA-OGN (15mol/L) attenuated ETA epression of HSV VSMCs in a time dependent manner ET-1 level in supernatant of HSV ECs significantly increased during 12～48 h and reached the peak at 48 h. ECCM stimulated the proliferation of VSMCs which was mimicked by ET-1. Phosphorimidon attenuated but did not eliminate the effect of ECCM. ETA-OGN (15mol/L) inhibited the proliferation induced by ECCM or ET-1 but had no significant effect on the proliferation of VSMCs cultured in DMEM without erium. CONCLUSION: HSV VSMCs can secrete different factors including ET-1, which play an important role in stimulating VSMC proliferation. ETA-OGN attenuates the proliferation of VSMCs from HSV induced by ECs. It suggests that OGN might be useful in the prevention and treatment of restenosis after angioplasty.

参考文献/References

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备注/Memo

备注/Memo:
收稿日期：2003-9-18
更新日期/Last Update: 2004-03-01