活化转录因子3在再狭窄静脉桥血管中的表达

Expression of activated transcription factor-3 in restenosis of vein graft

目的探讨再狭窄静脉桥血管与正常大隐静脉中活化转录因子3(ATF3)的表达以及机械牵张刺激对内皮细胞ATF3表达的影响。方法选择2017年7月首都医科大学附属北京安贞医院因静脉桥血管再狭窄需行二次冠状动脉旁路移植术(CABG)的患者及因冠状动脉粥样硬化性心脏病需行CABG患者各1例,CABG术中收集狭窄的静脉桥血管及患者自身正常的大隐静脉,每例样本分为3份,分别用于制成石蜡切片、制成冰冻切片、提取RNA。应用免疫组织化学法检测再狭窄静脉桥血管与正常大隐静脉血管ATF3的表达,应用实时荧光定量聚合酶链反应、蛋白质印迹法检测正常人脐静脉内皮细胞(HUVEC)及机械牵张刺激HUVEC中ATF3的表达。结果经免疫组织化学观察发现,ATF3在再狭窄静脉桥血管比正常人大隐静脉桥血管表达增高,且在内皮层表达明显增高。实时荧光定量聚合酶链反应检测经机械牵张刺激的HUVEC中ATF3表达高于正常HUVEC[(95±4)比(66±6)],差异有统计学意义(P=0.002)。经免疫印迹法检测经机械牵张刺激HUVEC中ATF3表达高于正常HUVEC[(0.43±0.03)比(0.15±0.02)],差异有统计学意义(P<0.001)。结论 ATF3可能在静脉桥血管再狭窄内皮功能失调过程中发挥重要作用。

Objective To explore the expression of activated transcription factor-3 （ATF3） in restenosis of vein graft and normal sapbenous vein; to analyze the effect of mechanical stretch stimulation on the expression of ATF3 in endothelial cells. Methods One patient undergoing secondary coronary artery bypass grafting （CABG） for restenosis of vein graft and 1 coronary heart disease patient undergoing primary CABG were enrolled in July 2017 at Beijing Anzhen Hospital, Capital Medical University. Restenosis of vein graft and normal saphenous vein were separated from patients to extract RNA; expressions of ATF3 in veins were tested by immunohistochemical technique; expressions of ATF3 in normal human umbilical vein endothelial cell（HUVEC） and HUVEC stimulated by mechanical stretch were tested by real-time fluorescence quantitative polymerase chain reaction and western blotting. Results Immunohistochemical test showed that ATF3 expression significantly increased in vein graft restenosis compared to that in normal saphenous vein. Polymerase chain reaction test showed that ATF3 expression in mechanical stretch-stimulated HUVEC was significantly higher than that in normal HUVEC [ （95 ± 4） vs （ 66± 6） ] （P = 0. 002）. Western blotting showed that ATF3 expression in mechanical stretch-stimulated HUVEC was significantly higher than that in normal HUVEC [ （0. 43 ± 0. 03） vs （0. 15 ± 0. 02） ] （P 〈 0. 001 ）. Conclusion ATF3 may play an important role in endothelial dysfunction of vein graft restenosis.