论文部分内容阅读
目的:探索表皮生长因子(EGF)对间充质干细胞(MSC)迁移的影响及其信号机制,提高口腔颌面部损伤修复的能力。方法:采用全骨髓贴壁法培养MSC,通过流式细胞仪分析其表面标记(CD45、CD34、CD90、CD29)以及成骨、成脂肪等多向诱导分化潜能鉴定其特征。采用MTT法分析不同浓度的EGF对MSC增殖的影响,采用Transwe ll体外迁移体系观察不同浓度的EGF对MSC迁移的影响,随后以MEK阻断剂PD98059(50μM)、p38 MAPKSB203580(30μM)、PKC非选择性阻断剂Straurosporine(10 mM)及磷脂酶C阻断剂U73122(10μM)等分别处理MSC,观察合适浓度的EGF影响MSC迁移与PKC/ERK 1/2途径等途径的关系。结果:培养的MSC表现出CD90、CD29强阳性,具有成骨、成脂肪等多向分化能力;EGF促进了MSC迁移,U73122、Straurosporine及PD98059等可明显抑制了EGF所诱导的MSC迁移效应。结论:EGFF引起的MSC迁移效应与PLC/PKC/ERK 1/2途径相关,能有效促进颌面部损伤的修复。
Objective: To explore the effect of epidermal growth factor (EGF) on the migration of mesenchymal stem cells (MSCs) and its signal mechanism, and to improve the ability of oral and maxillofacial injury repair. Methods: MSC were cultured with whole bone marrow adherent method. The surface markers (CD45, CD34, CD90, CD29) and the multi-directional differentiation potential of osteoblast and adipose tissue were identified by flow cytometry. MTT assay was used to analyze the effect of different concentrations of EGF on the proliferation of MSCs. Transwell migration in vitro was used to observe the effect of different concentrations of EGF on the migration of MSC. The effects of MEK inhibitor PD98059 (50μM), p38 MAPKSB203580 (30μM) The selective blockers Straurosporine (10 mM) and the phospholipase C blocker U73122 (10 μM) were used to treat MSCs respectively. The effect of appropriate concentration of EGF on the migration of MSC and the pathway of PKC / ERK 1/2 were observed. Results: The cultured MSCs showed strong positive of CD90 and CD29, with the ability of osteoblast differentiation and adipose differentiation. EGF promoted the migration of MSC. U73122, Straurosporine and PD98059 could obviously inhibit the migration of MSCs induced by EGF. Conclusion: The migration effect of EGFF induced by MSC is related to the pathways of PLC / PKC / ERK 1/2, which can effectively promote the repair of maxillofacial injuries.