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目的血小板衍生生长因子BB(PDGF-BB)是介导血管新生内膜形成最重要的生长因子之一,本研究探讨PDGF-BB激活细胞外信号调节激酶1/2(ERK1/2)参与血管新生内膜的形成。方法在整体实验上,将24只250g健康雄性SD大鼠分为3组:A组为假手术组;B组为颈动脉球囊损伤组;C组为颈动脉球囊损伤干预组(ERK1/2抑制剂PD98059)。建立颈动脉球囊损伤模型后,经血管外膜途径局部给予PD98059抑制ERK1/2激活,苏木精-伊红(HE)染色分析损伤血管形态学变化,免疫荧光检测巨噬细胞浸润。原代培养外膜成纤维细胞,Western blot检测细胞ERK1/2的激活,划痕实验观察细胞迁移变化。结果损伤组新生内膜面积和内膜中膜比大于干预组[(0.149±0.012)比(0.077±0.021)mm2,1.137±0.088比0.682±0.141,均P<0.01]。损伤组管腔面积小于干预组[(0.249±0.021)比(0.314±0.016)mm~2,P<0.05]。同时,PD98059能明显改善新生内膜形成,伴随巨噬细胞浸润明显减少。20μg/L PDGF-BB瞬时激活ERK1/2在5min时磷酸化ERK1/2表达最高,并且ERK1/2的激活能够被PD98059所抑制。PD98059也抑制PDGF-BB促进外膜成纤维细胞的迁移。结论经外膜干预ERK1/2激活可以改善损伤诱导的新生内膜形成,可能是通过抑制PDGF-BB介导的外膜成纤维细胞的迁移。
OBJECTIVE: Platelet-derived growth factor BB (PDGF-BB) is one of the most important growth factors that mediate angiogenesis. In this study, PDGF-BB is involved in the activation of extracellular signal-regulated kinase 1/2 (ERK1 / 2) Intimal formation. Methods Twenty-four healthy male Sprague Dawley rats weighing 250g were divided into 3 groups: group A was sham operation group, group B was carotid artery balloon injury group, group C was carotid artery balloon injury intervention group (ERK1 / 2 inhibitor PD98059). The model of carotid artery balloon injury was established. PD98059 was administered via the adventitia of the blood vessel to inhibit the activation of ERK1 / 2. The morphology of injured vessels was analyzed by hematoxylin - eosin (HE) staining and the infiltration of macrophages was detected by immunofluorescence. Primary culture of adventitial fibroblasts, Western blot detection of ERK1 / 2 activation, scratch test observed changes in cell migration. Results The neointimal area and intima-media ratio in the injured group were significantly higher than those in the intervention group [(0.149 ± 0.012) vs (0.077 ± 0.021) mm2,1.137 ± 0.088 vs 0.682 ± 0.141, both P <0.01]. The lumen area in the injury group was smaller than that in the intervention group [(0.249 ± 0.021) vs (0.314 ± 0.016) mm ~ 2, P <0.05]. At the same time, PD98059 can significantly improve neointimal formation, accompanied by significantly reduced macrophage infiltration. At 20μg / L PDGF-BB, the phosphorylation of ERK1 / 2 was the highest at 5min, and the activation of ERK1 / 2 was inhibited by PD98059. PD98059 also inhibits PDGF-BB to promote the migration of outer membrane fibroblasts. Conclusion Activation of ERK1 / 2 by the outer membrane can improve damage-induced neointima formation possibly by inhibiting the migration of outer membrane fibroblasts mediated by PDGF-BB.