七叶皂苷对大鼠体外培养神经元的影响及其与NGF信号系统的关系  

Effect of Aescin on Cultured SD Rat Neurons and Its relationship with NGF Signal System

作　　者：李俊彦[1] 罗涛[2] 杨金伟[3] 王东艳[2] 茹金[2] 李力燕[2] 刘俊[1] 

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

出　　处：《昆明理工大学学报（自然科学版）》2014年第3期83-88,共6页Journal of Kunming University of Science and Technology(Natural Science)

基　　金：国家自然科学基金项目(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项目(2011DH011)

摘　　要：为了探讨七叶皂苷对SD大鼠大脑皮质体外培养神经元生长的影响及其与神经生长因子(NGF)信号通路的关系.取新生SD大鼠胚胎的大脑皮质,在24或96孔板中进行原代培养神经元,随机分为正常组(A组)、对照组(B组)、七叶皂苷组(C组),其中C组在细胞长至80%融合度时加入10 mg/mL七叶皂苷(采用生理盐水稀释),B组加入等量生理盐水,A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均采集图像,之后采用逆转录-聚合酶链式反应(Reverse Transcription polymerase Chain Reaction,RT-PCR)技术检测NGF、TrkA mRNA的表达变化.对96孔板中各组细胞各时间点采用MTT检测神经元生长活力.结果显示A组和B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A组及B组均有随时间点延长,三个指标上调的时间点(P<0.05),C组细胞数、胞体面积、突起长度及细胞活力均较A组及B组升高(P<0.05).NGF及TrkA mRNA的表达在A组和B组在各时间点无明显差异(P>0.05),C组各时间点较A组和B组上调(P<0.05),但随时间的延长,各组组内比较无统计学意义(P>0.05).表明七叶皂苷可促进SD大鼠脑源性神经元的存活、生长,其机制可能是通过上调NGF及其受体TrkA的表达来实现.This paper aims to explore the effect of Aescin on cerebral cortex SD rats cultured neurons growth in vitro and its relationship with NGF signaling system. Cerebral cortex of newborn SD rat embryos is collected.Neurons are cultured in 24 or 96 well plate by primary culture and then randomly divided into normal group （group A）,control group （group B）,Aescin group （group C）. Group C is injected with 10 mg/mL Aescin （diluted with normal saline）when the fusion degree reaches 80%;same amount of normal saline is employed in group B,while group A has no treatment. Each group is then divided into 24h and 48h sub-groups. Images of are captured. RT-PCR is then adopted to examine the expression of NGF and TrkA mRNA. MTT is used to detect the neuronal growth vigor in each group at each time point in 96 well plate. There is no difference in group A and B at each time point in cell numbers,body area,neurite length and viability （P〉0.05 ）,but with the time prolonged,the indexes will rise in both groups （P〈0.05 ）. The corresponding indexes in group C are all higher than the other two groups. RT-PCR shows that there is no significant difference in expression of NGF and TrkA mRNA in group A and B at each time point （P〉0.05 ）,while it increases at each time point in group C （P〈0.05 ）,but with the extension of time,it has no statistical significance （P〉0.05 ）within each group. It is,therefore,proven that Aescin can promote the survival and growth of SD rat brain derived neurons,and the mechanism might be through up-regulating the expression of NGF and its receptor TrkA.

关 键 词：七叶皂苷 大鼠 神经元 神经生长因子 酪氨酸激酶A 

分 类 号：R651[医药卫生—外科学]