摘要
目的探讨血管紧张素II(Ang II)1型受体相关蛋白(ATRAP)通过Ang II 1型受体(AT1R)调节血管平滑肌细胞(VSMCs)增殖及血管内膜增生的作用机制。方法将ATRAP cDNA转至pcDNA3载体,然后转染至来源于成年SD大鼠胸主动脉的VSMCs。VSMCs内DNA合成,细胞外信号调控激酶(ERK)及ERK磷酸化的表达分别通过3H胸腺嘧啶核苷的合成和Western blot方法检测。利用球囊损伤血管的模型,观察转染和未转染ATRAP cDNA的12周雄性SD大鼠的血管形态学变化。结果在Ang II刺激VSMCs 48h后,ATRAP过表达可以抑制Ang II诱导的3H胸腺嘧啶核苷的合成(P<0.05)。在VSMCs内,Ang II的刺激增加了ERK的磷酸化,大约60min达到峰值。ATRAP的过表达使磷酸化ERK的活性显著降低(P<0.05),并明显抑制了损伤动脉的新生内膜增生。结论 AT1受体介导的ERK活性在Ang II调节的VSMCs增殖中起重要作用。ATRAP对VSMCs增殖和新生内膜形成的抑制作用可能是由于其干扰AT1受体介导的ERK活性。
Objective To investigate the effect of angiotensin II (Ang II) type 1 (AT1) receptor-associated protein (ATRAP) on vascular smooth muscle cells (VSMCs) proliferation and neointimal formation mediated by AT1 receptor. Methods ATRAP cDNA was subcloned into pcDNA3 vector, and was transfected into VSMCs, which were isolated from thoracic aorta of adult Sprague-Dawley (SD) rats. DNA synthesis, extracellular signal-regulated kinase (ERK) and phospho-ERK expression in VSMCs were analyzed by measurement of 3H thymidine synthesis and Western blot respectively. Morphological changes were observed in the balloon injured artery of 12-week male SD rats with or without transfection of ATRAP cDNA. Results ATRAP overexpression in VSMCs inhibited Ang II-induced 3H thymidine synthesis after 48 h stimulation (P<0.05). In VSMCs, Ang II stimulation increased the phosphorylation of ERK, which reached the peak at 60min. ATRAP overexpression in injuried arteries significantly decreased the activation of phospho-ERK and inhibited neointimal formation (P<0.05). Conclusion The AT1 receptor-mediated activation of ERK plays an essential role in Ang II-regulated VSMCs proliferation. The inhibitory effects of ATRAP on VSMCs proliferation and neointimal formation might be due to interfering with AT1 receptor-mediated activation of ERK.
出处
《中国血管外科杂志（电子版）》
2013年第4期222-224,232,共4页
Chinese Journal of Vascular Surgery(Electronic Version)
基金
国家自然科学基金面上项目(81270393)
