神经生长因子对人脐静脉内皮细胞在体外形成管腔样结构的影响  

Effect of nerve growth factor on the formation of lumen-like structures in cultured human umbilical vein endothelial cells

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,大连116044 [2]沈阳军区总医院心内科心血管研究所

出　　处：《中华老年心脑血管病杂志》2009年第1期48-51,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的探讨神经生长因子(NGF)对人脐静脉内皮细胞(HUVECs)在体外形成管腔样结构(LLSs)的影响及其信号机制。方法将接种在基质胶上的HUVECs分为空白对照组、NGF组、NGF中和抗体组、对照IgG组、NGF中和抗体+NGF组、对照IgG+NGF组,以相差显微镜直接观察LLSs形态,并通过二脒基苯基吲哚/鬼笔环肽免疫荧光染色观察LLSs的细胞组成。为证实NGF促LLSs形成的信号通路,向培养液中分别预先加入丝裂原活化蛋白激酶3个主要信号通路的特异性抑制剂(PD98059、SB203580及SP600125),观察上述3个信号通路对NGF促LLSs形成的影响,并应用Western blot检测NGF及c-Jun N末端激酶(JNK)特异性抑制剂SP600125对HUVECs JNK表达及活性的影响。结果 NGF组LLSs的平均面积较空白对照组明显增加(P<0.01)。NGF中和抗体+NGF组LLSs的平均面积较对照IgG+NGF组明显减少(P<0.01);NGF中和抗体组LLSs的平均面积较对照IgG组亦明显减少(P<0.01)。NGF可激活HUVECs的JNK信号通路,SP600125可显著减少NGF诱导的LLSs形成(P<0.01)。PD98059及SB203580对NGF诱导的LLSs形成均无明显影响(P>0.05)。结论NGF能促进HUVECs在体外形成LLSs,可能与激活JNK信号通路有关。Objective To investigate the molecular mechanism of effect of nerve growth factor （NGF） on the formation of lumen-like structures （LLSs） in human umbilical vein endothelial cells （HUVECs） in vitro. Methods Cultured HUVECs were divided into following groups：control, NGF, NGF neutralizing antibody, control IgG, NGF neutralizing antibody ＋ NGF, control IgG＋NGF. The configuration of LLSs was observed under phase contrast microscope and the cellular composition of LLSs was investigated using immunofluorescence staining. To detect the signaling pathway by which NGF promoted the formation of LLSs,inhibitors of three main members of MAPK （PD98059, SB203580, and SP600125） were pre-added into the medium. Western blot was performed to observe the effect of NGF or SP600125 on the expression and activity of c-Jun N terminal kinase（JNK）. Results The average LLSs area of NGF group significantly increased compared with that of control group. NGF-neutralizing antibody markedly blocked LLSs formation in the presence of PBS or NGF. NGF could activate JNK in HUVECs and SP600125 significantly reduced NGF induced LLSs formation. Conclusions Formation of LLSs in HUVECs promoted by NGF may be associated with activation of JNK signaling pathway.

关 键 词：神经生长因子 脐静脉 内皮细胞 信号传导 丝裂原激活蛋白激酶类 

分 类 号：R363[医药卫生—病理学]