摘要
目的 探讨细胞外信号调节激酶(ERKs)在自体移植静脉中表达的变化规律。方法Wistar大鼠 80只,建立自体静脉移植模型,术后随机分为 6、24 h,3、7 d,2、4、6、8周等 8组,半定量逆转录-聚合酶链反应(RT-PCR)检测移植血管中 ERK1 mRNA表达,Western蛋白印迹定量ERK1/2的蛋白产物以及磷酸化蛋白产物的表达,原位杂交定位 ERK1 mRNA、免疫组织化学方法检测 ERK和增殖细胞核抗原(PCNA)的蛋白产物表达。结果 静脉移植后 6 h,ERK1 mRNA表达明显增强,与正常静脉比较差异有显著性(P<0.01),在移植7 d表达达高峰(33.2±14.2)％,与其余时点比较差异有显著性(P<0.01),4周后恢复至正常水平。Western蛋白印迹提示 ERK1/2在术后1～2周达高峰。阳性表达多定位于血管平滑肌细胞(VSMCs),ERK1与PCNA表达呈正相关(r=0.759 6,P<0.01)。结论 ERKs信号转导通路的激活可能是内膜增生的关键环节,可能成为防治移植血管狭窄、闭塞新的靶点。
Objective To investigate the gene expression of extracellular signal-regulated kinases
(ERKs) in autogenous vein graft in rats. Methods Autogenous vein graft model was established by
transplanting the right jugular vein to infrarenal abdominal aorta in 80 Wistar rats. Vein graft samples
were harvested 6 h, 24 h, 3 days, 7 days, 2 weeks, 4 weeks, 6 weeks and 8 weeks after surgery. The ex-
pression of ERK1 mRNA was detected by reverse transcription-PCR and in situ hybridization. Western
blot and immunohistochemistry methods were used to detect the protein production and phosphorylation
of ERKs. Proliferating cell nuclear antigen (PCNA) also was studied. Results The expression of ERK1
mRNA was increased quickly after surgery and reached the peak on 7th day [ (33. 2 ± 14. 2)%, P < 0.01
as compared normal other time points] and returned to normal levels 4 weeks after surgery. The expres-
sion of ERK1/2 detected by western blot reached the peak during 1 to 2 weeks and was decreased gradu-
ally to normal level 6 weeks after surgery. The positive cells were mostly vascular smooth muscle cells
(VSMCs) located in media and intimal hyperplasia of vein graft. There was a positive relationship be-
tween ERK1 and PCNA(r = 0. 759 6, P < 0. 01). Conclusion The activation of ERKs present in auto-
genous vein graft may become a new target for the therapy of intimal hyperplasia and stenosis after vein
graft.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2003年第12期1111-1113,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(30371401
30000163)
