摘要
目的探讨藏药旺拉提取物CE在Aβ25-35所致大鼠前额叶神经元损伤中的作用。方法运用大鼠前额叶神经细胞原代培养技术,采用Aβ25-35诱导建立神经细胞损伤模型。实验共分为6组:正常对照组,Aβ25-35模型组,CE高、中、低浓度预处理组(CE终浓度分别为10,1,0.1 mg·ml-1)和CE组(终浓度为10 mg·ml-1)。免疫组化法鉴定大鼠前额叶神经细胞纯度;镜下观察神经细胞形态学变化;MTT法测定细胞活性;TUNEL法检测细胞凋亡情况;免疫组化法检测大鼠前额叶神经细胞中微管蛋白β-tubulin Ⅲ蛋白的形态结构及表达。结果培养的大鼠前额叶神经细胞纯度达95%以上;形态学观察显示:与模型组比较,CE预处理组神经细胞损伤程度明显减轻;24 h的CE高、中浓度预处理CE发挥着对Aβ25-35所致大鼠前额叶神经元损伤的保护作用,MTT法显示CE中高浓度预处理组细胞活性(0.783,0.667 3)比模型组细胞活性(0.603 8)明显升高(P<0.001或P<0.01);TUNEL法显示CE10 mg·ml-1高浓度预处理组细胞凋亡阳性率(11.1%)明显低于模型组细胞凋亡阳性率(32.1%)(P<0.001);CE10 mg·ml-1高浓度预处理组微管蛋白β-tubulinIII结构正常,表达量明显高于模型组。结论藏药旺拉提取物CE对Aβ25-35所致大鼠前额叶神经元损伤具有一定的保护作用,其作用与其维护神经元中微管蛋白β-tubulin III的正常形态及表达有关。
Objective To study the effect of Coeloglossum viride var.bracteatum extract（CE） against the injury induced by β-amyloid protein（Aβ） in primary cultivated rat prefrontal cortex（PFC） neurons.Methods The neuron purity of PFC was indentified by immunohistochemical β-tubulin III and DAPI method.The change of cell morphology was observed by microscope,the protective effect of CE on Aβ25-35 induced neuron injury was observed by MTT and TUNEL.The expression and structure of β-tubulin III were detected by immunohistochemical method.Results There were 95% neurons in the cultivated rat primary prefrontal tissue.Compared with model group,the damage of PFC neurons in CE treatment groups was slighter than that in model group;24 hours pretreatment with 10 mg·L-1,1 mg·L-1 CE exerted the protective effect on rat primary PFC neuron injury Aβ25-35 treatment,the viability of PFC neurons in CE high and medium treatment groups（0.783,0.667 3） was higher than that in model group（0.603 8）（P0.001 or P0.01）;Aβ25-35 could increase cell death in rat PFC neurons,the rate of masculine in CE high treatment groups（11.1%） was obviously lower than that in model group（32.1%） with TUNEL method;CE could increase the expression of β-tubulin III and maintained β-tubulin Ⅲ srtucture in the injured rat primary PFC neurons induced by Aβ25-35.Conclusion CE has protective effects against the injury induced by Aβ25-35 in rat primary prefrontal cortex neurons to some existence,and its mechanism may be related with the increase of β-tubulin III expression and maintainment of the injured β-tubulin III structure.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2011年第5期1065-1068,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(No.30701138)
中央民族大学985资助项目(No.CUN985-3-3)
