摘要
目的:观察凝血酶对骨髓来源内皮祖细胞(EPCs)增殖能力的影响并探讨其机制。方法:密度梯度离心法从大鼠骨髓分离EPCs,培养7 d后收集贴壁细胞并加入不同浓度凝血酶(10-2,10-1,1,10,100 U/mL)干预一定时间(6,12,24,48 h)。CCK-8试剂盒检测EPCs增殖。实时定量PCR检测凝血酶刺激以及NF-κB抑制剂预处理以后EPCs的VEGF mRNA表达变化,ELISA法检测其分泌量变化。结果:凝血酶干预组的细胞增殖数均高于对照组,10 U/mL凝血酶作用24 h对内皮祖细胞数量影响最为显著(P<0.05)。与对照组相比,凝血酶刺激后VEGFmRNA表达及分泌量均增高,使用NF-κB抑制剂预处理后可以抑制凝血酶的这种作用。结论:凝血酶可以通过NF-κKB途径上调VEGF的表达,促进EPCs增殖。
Objective: To investigate the effects of thrombin on proliferation of bone-marrow de- rived endothelial progenitor cells (EPCs)and the underlying mechanism. Methods: EPCs were acquired from the bone marrow of SD rats by density gradient centrifugation. After 7 days of culture, EPCs were incubated with thrombin in different concentrations(10-2,10 -1, 1,10,100 U/mL)for different time points (6,12,24,48 h). EPCs proliferation was measured by Cell Counting Kit-8 Real time PCR was used to measure VEGF mRNA expression of EPCs after intervention by thrombin or pretreatment with NFkB inhibitor. ELISA assay was used to measure VEGF secretion. Results: EPCs proliferation was significant higher in the thrombin intervention group compared with control group, it reached a maximum at 10 U/mL after 24h of intervention (P〈0.05). Thrombin up-regulated VEGF mRNA level and increased the se cretion, which process can be blocked by NFkB inhibitor. Conclusion: Thrombin could induce the proliferation of EPCs via increasing the expression of VEGF through NFkB pathway.
出处
《海南医学院学报》
CAS
2010年第1期1-3,6,共4页
Journal of Hainan Medical University
基金
国家自然科学基金(30770849)~~
关键词
内皮祖细胞
细胞增殖
凝血酶
骨髓
Endothelial progenitor cells
Cell proliferation
Thrombin
Marrow