外源性bFGF对深度烫伤大鼠创面血管内皮细胞增殖与迁移的影响

EFFECT OF EXOGENOUS BASIC FIBROBLAST GROWTH FACTOR ON PROLIFERATION AND MIGRATION OF ENDOTHELIAL CELLS OF PARTIAL THICKNESS SCALD IN RATS

目的 观察大鼠深 度烫伤创面敷用 b FGF后肉芽组织中血管增殖与迁移情况 ,以及相关信号蛋白的表达情况。 方法 Wistar大鼠 133只随机分为正常对照 A组 (n=7)、单纯烫伤 B组 (n=4 2 )、b FGF治疗 C组 (n=4 2 )、c- fos抗体 D组 (n=4 2 )。利用大鼠 30 %深 度烫伤模型 ,于伤后 3、6小时 ,1、3、7、 14和 2 1天取创面皮肤标本 ,运用免疫组织化学染色 ,检测真皮内血管形成的情况 ,原位杂交和免疫组织化学技术观察血管内皮细胞增殖细胞核抗原 (proliferation cell nuclear antigen,PCNA)、黏着斑蛋白激酶 (focal adhesion kinase,FAK)及 c- fos的表达情况。免疫荧光手段观察细胞外信号调节激酶 (extracellularsignal- regulated kinase,ERK)在血管形成中的激活情况。 结果 B、C组于伤后 7天血管内皮细胞增殖明显 ,14天后 FAK的表达也显著增加。损伤后 3～ 6小时 ERK的表达增强 ,随后 1～ 3天 ,c- fos亦发生相对应的变化。各时间点 D组血管内皮细胞 PCNA和 FAK的表达均减弱 ,肉芽组织中微血管的数量低于 B组。敷用外源性的 b FGF后 ,血管内皮细胞的增殖与前者变化不显著 ,但 FAK的表达较 A组增强明显 ,ERK的表达也显著增加 ,特别是 c- fos也在伤后 1～ 3天呈增强趋势。 结论 使用外源性 b

Objective To observe the proliferation and migration of endothelial cells after 30% total burn surface area (TBSA) of deep partial thickness scald, and the effect of basic fibroblast growth factor (bFGF) on angiogenesis during wound healing.Methods A total of 133 male Wistar rats were divided randomly into normal control (n=7), injured control group (n=42), bFGF group (n=42) and anti-c-fos group (n=42). The apoptosis expression of fibroblasts was determined with in situ hybridization and the changes of proliferation cell nuclear antigen(PCNA), focal adhesion rinase(FAK), c-fos and extracellular signal-regulated kinase(ERK) proteins expression were detected with immunohistochemistry staining technique after 3 hours, 6 hours, 1 day, 3 days, 7 days, 14 days and 21 days of scald.Results In injured control group and bFGF group, the proliferation rate of the vascular endothelial had evident changes 7 days and 14 days after scald; the expression of FAK was increased 14 days after scald. ERK proteins expression was different between injury control group and bFGF group at initial stage after scald. Stimulation of ERKs by bFGF led to up-regulation of c-fos and strong expression of FAK. Conclusion Exogenous bFGF extended the influence on wound healing process by ERK signaling pathway, affecting migration cascade of vascular endothelial cell. The oncogene proteins play an important role on accelerating angiogenesis during wound healing.