CAJ | 학술논문

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．
위료탐토칠협조감대SD대서대뇌피질체외배양신경원생장적영향급기여신경생장인자（NGF）신호통로적관계．취신생SD대서배태적대뇌피질，재24혹96공판중진행원대배양신경원，수궤분위정상조（A조）、대조조（B조）、칠협조감조（C조），기중C조재세포장지80％융합도시가입10 mg/mL칠협조감（채용생리염수희석），B 조가입등량생리염수，A조불작임하처리，각조우분위24 h、48 h아조．대24공판중각조세포각시간점균채집도상，지후채용역전록-취합매련식반응（Reverse Transcription polymerase Chain Reaction，RT -PCR）기술검측 NGF、TrkA mRNA적표체변화．대96공판중각조세포각시간점채용MTT검측신경원생장활력．결과현시A조화B조재각시간점세포수、포체면적、돌기장도급세포활력무명현차이（P＞0.05），단A조급B조균유수시간점연장，삼개지표상조적시간점（P＜0.05），C조세포수、포체면적、돌기장도급세포활력균교A조급B조승고（P＜0.05）. NGF급TrkA mRNA적표체재A조화B조재각시간점무명현차이（P＞0.05），C 조각시간점교A조화B 조상조（P＜0.05），단수시간적연장，각조조내비교무통계학의의（P＞0.05）.표명칠협조감가촉진SD대서뇌원성신경원적존활、생장，기궤제가능시통과상조NGF급기수체TrkA적표체래실현．
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.