灯盏花素干预体外培养大鼠脑皮质神经元的存活与生长  

Effect of Breviscapine on the survival and growth of rat cortical neurons cultured in vitro

作　　者：李俊彦[1] 王东艳[2] 杨金伟[3] 赵楠[1] 李力燕[2] 程敬茹 茹金[2] 郭建辉[3] 刘俊[1] 

机构地区：[1]昆明市第一人民医院神经外科,云南省昆明市650031 [2]昆明医科大学神经科学研究所,云南省昆明市650500 [3]云南省第一人民医院普外二科,云南省昆明市650032

出　　处：《中国组织工程研究》2014年第20期3196-3201,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81260075;31260253);云南省科技厅-昆明医科大学应用基础研究联合专项(2012FB002;2012FB092);云南省肿瘤转化医学工程技术研究中心(2011DH011)~~

摘　　要：背景:神经生长因子在维持中枢神经系统神经细胞的存活、分化,促进其生长、发育,维持其功能方面具有不可替代的作用。目的:观察灯盏花素对SD大鼠大脑皮质体外培养神经元生长的影响,并初步探讨其机制。方法:取新生SD大鼠胚胎,取其大脑皮质后,对神经元进行原代培养,随机分为3组,其中灯盏花素组加入10 g/L灯盏花素,对照组加入等量生理盐水,正常组不作任何处理,各组又分为24 h、48 h亚组。对24孔板中各组细胞各时间点采集图像,之后采用RT-PCR技术检测神经生长因子、酪氨酸激酶A mRNA的表达变化。对96孔板中各组细胞不同时间点采用MTT检测神经元生长活力。结果与结论:正常组和对照组在各时间点细胞数、胞体面积、突起长度及细胞活力无明显差异(P>0.05),但正常组及对照组均有随时间点延长,3个指标均上调(P<0.05),灯盏花素组细胞数、胞体面积、突起长度及细胞活力均较正常组及对照组升高(P<0.05)。神经生长因子及TrkA mRNA的表达在正常组和对照组在各时间点无明显差异(P>0.05),灯盏花素组各时间点较正常组和对照组上调(P<0.05)。提示灯盏花素促进SD大鼠脑源性神经元的存活、生长,其机制可能是通过上调神经生长因子及其受体TrkA的表达发挥作用。BACKGROUND：Nerve growth factor plays an irreplaceable role on the survival, differentiation and maintenance of the central nervous system cells, also promote the growth and development of these cells. OBJECTIVE：To explore effect of Breviscapine on the growth of in vitro cultured neurons in cerebral cortex of Sprague-Dawley rats, and preliminarily investigate the action mechanism. METHODS：The cerebral cortex of newborn Sprague-Dawley rat embryos was col ected, the neurons were primary cultured and randomly divided into three groups：normal control group （no treatment）, control group （neurons were cultured with normal saline）, Breviscapine group （neurons were cultured with 10 g/L Breviscapine）. Furthermore each group was assigned into 24-hour and 48-hour subgroups. Images were captured from the 24-wel plates at each time points. RT-PCR was applied to examine the expression of nerve growth factor and TrkA mRNA. MTT was used to detect the neuronal growth at each time point. RESULTS AND CONCLUSION：There was no difference between normal control group and control group in the cellcounts, cellbody area, neurite length and viability （P〉0.05）, as the time prolonged, al data were raised in the＆nbsp;two groups （P0.05）. At each time point, these data in Breviscapine group were increased compared with normal control group and control group （P〈0.05）. Breviscapine can promote the survival and growth of brain-derived neurons in Sprague-Dawley rats, and the mechanism might depend on the up-regulating expression of nerve growth factor and its receptor TrkA.

关 键 词：组织构建 组织工程 灯盏花素 神经元 神经生长因子 酪氨酸激酶 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]