神经生长因子启动肾上腺髓质细胞功能冗余性致肾上腺素释放障碍的研究  

Dysfunction of releasing adrenaline in primary cultured AMCC due to functional reduncdency primed by nerve growth factor.

作　　者：汪俊[1] 胡成平[2] 冯俊涛[2] 

机构地区：[1]江西省人民医院呼吸内二科,南昌330006 [2]中南大学湘雅医院呼吸内科,长沙410008

出　　处：《中国实用内科杂志》2009年第4期336-338,共3页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金资助项目(30570802)

摘　　要：目的探讨神经生长因子(NGF)等对原代培养的肾上腺髓质嗜铬细胞生物行为学影响的发生机制。方法采用单细胞消化法原代培养肾上腺髓质细胞,密度梯度离心和差速贴壁法分离和纯化肾上腺髓质细胞。相差显微镜、电镜观察外源性NGF、P物质(SP)作用下肾上腺髓质嗜铬细胞形态和超微结构的改变,酶联免疫吸附试验(ELISA)法检测培养上清液中肾上腺素和去甲肾上腺素浓度的变化。结果NGF干预2 d后,相差显微镜下可见肾上腺髓质细胞膜表面渐渐长出长丝状突起。电镜下NGF干预后可见肾上腺髓质嗜铬细胞表面丰富的杵状和绒毛状突起,近细胞膜处可见小囊泡形成,细胞浆内线粒体丰富,结构较模糊,部分嵴丢失。ELISA检测发现经NGF干预后培养上清液中肾上腺素浓度较正常组明显降低(P<0.05),去甲肾上腺素浓度没有明显改变。SP干预后肾上腺素和去甲肾上腺素浓度没有明显改变。结论NGF通过启动肾上腺髓质细胞功能冗余性使其表型和功能发生改变致肾上腺素合成和释放障碍。Objective To investigate the possible mechanism of the biologic ethology effects of NGF and SP （ substance P） on primary cultured adrenaline medullary chromaffin cells （AMCC）. Methods To establish primary cultured AMCC by means of enzyme digestion and purify the cells by means of isopycnic gradient centrifugation and differential plating. To observe the morphological and uhrastructural changes after the addition of NGF and SP, and detect the concentration of adrenaline and noradrenaline in serum by ELISA. Results Confervaceous processes could be observed after 2 d of addition of NGF to the culture and the processes strenched longer as days went by the observation of electron microscopy, there are some drumstick-like and villiform processes in the cell membrane and some vesiculation be observed near the cell membrane of the primary cultured AMCC cells after the addition of NGF. The bioblast was abundant but the structure was not clear in the intracytoplasm and the concentration of adrenaline were decreased （ P 〈 0.05 ） after the addition of NGF, while the contration of noradrenalin were not changed. The concentration of adrenaline and noradrenalin were also not changed after the addition of SP. Conclusion NGF could prime the functional reduncdancy of AMCC,which would lead to the Dysfunction of releasing adrenaline due to phenotypic and functional changes.

关 键 词：神经生长因子 肾上腺髓质嗜铬细胞 功能冗余性 肾上腺素 

分 类 号：R5[医药卫生—内科学]