神经生长因子对体外培养的小鼠主动脉环血管新生的影响及TrkA的关键作用  

Effect of nerve growth factor on angiogenesis of in vitro cultured murine aorta ring and key role of TrkA

作　　者：齐岩梅[1] 韩雅玲[2] 康建[2] 李娜[2] 田孝祥[2] 闫承慧[2] 

机构地区：[1]大连医科大学研究生院,辽宁大连116044 [2]沈阳军区总医院心内科心血管研究所,辽宁沈阳110016

出　　处：《心脏杂志》2009年第1期33-37,共5页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(30370526)

摘　　要：目的探讨神经生长因子(NGF)对体外培养的小鼠主动脉环血管新生的影响。方法①将小鼠主动脉环在三维基质胶中以含不同浓度的NGF的无血清培养基培养不同天数后,对出芽的细胞进行免疫荧光染色细胞学鉴定,或在倒置显微镜下观察、摄像,并以ImagePro图像分析软件计算每高倍视野中平均出芽的面积。②向培养基中预先加入抗NGF的中和抗体或血管内皮生长因子受体2(VEGFR2)拮抗剂SU5416,观察NGF或VEGF对出芽性血管新生的影响。③向培养基中预先加入酪氨酸受体激酶A(TrkA)受体拮抗剂K252a,观察NGF是否可通过TrkA受体促血管新生。结果①外源性NGF呈浓度依赖性促进基质胶中培养的小鼠主动脉环出芽。出芽的细胞上CD31的表达阳性。抗NGF的中和抗体能明显阻断NGF诱导的出芽性血管的新生。②SU5416可显著减少VEGF诱导的血管新生;但对NGF诱导的血管新生无明显影响。③K252a可明显抑制NGF诱导的血管新生。结论NGF可通过TrkA受体促进小鼠主动脉环血管新生,为促血管新生治疗方案的选择提供了新的思路。AIM To explore the effect of nerve growth factor （NGF） on angiogenesis of in vitro cultured murine aorta ring. METHODS （1）Murine aorta rings were cultured in 3D Matrigel in serum-free medium in the presence of various concentrations of NGF for different time periods. The identification of sprouted vascular endothelial cells was confirmed by immunofluorescent staining. The configuration of sprouting was observed and photographed under phase contrast microscope and the average sprouting area per high power field was measured by Image Pro software and analyzed statistically with SPSS13.0 software. （2）NGF neutralizing antibody or vascular endothelial growth factor （VEGF） receptor 2 （ R2 ） antagonist SU5416 pretreatment was used to investigate the effect of NGF or VEGF on angiogenesis. （3）Tyrosine receptor kinase A （TrkA） antagonist K252a pretreatment was used to investigate whether NGF exerted its angiogenic effect through its receptor TrkA. RESULTS （1）Exogenous NGF promoted endothelial cell sprouting from murine aorta rings cultured in Matrigel in dose-dependent manner. Sprouted cells expressed CD31. Anti- NGF neutralizing antibody markedly blocked NGF-induced sprouting angiogenesis. （2）SU5416 significantly reduced VEGF-induced angiogenesis but had no significant effect on NGF-induced sprouting. （3）K252a significantly inhibited NGF-induced angiogenesis. CONCLUSION NGF promotes angiogenesis of murine aorta ring through activating TrkA receptor, which may provide some new insights into pro-angiogenic therapy.

关 键 词：神经生长因子 主动脉环 血管新生 小鼠 

分 类 号：Q593.2[生物学—生物化学]