摘要
目的探讨益肾化浊方对阿尔兹海默病模型快速老化鼠(SAMP)8空间记忆能力、海马神经元形态学的影响及其作用机制。方法将48只9月龄健康雄性SMAP8采用双盲法随机分为SAMP8组、安理申组、益肾化浊方组,每组16只,另取16只SAMR1作为对照组。各组小鼠分别灌胃给予安理申(30 mg/kg)、益肾化浊方(20 g/kg)或等体积0.9%生理盐水,2次/d,连续14 w。分别采用Morris水迷宫检测空间学习记忆能力,尼氏染色观察细胞凋亡情况,电镜观察神经元超微结构,RT-PCR、Western印迹检测β-淀粉样蛋白前体(APP)、早老蛋白(PS1)基因和GSK-3β蛋白表达。结果与对照组比较,SAMP8组游泳总路程、逃避潜伏时间、海马CA1区神经元凋亡数目和APP、PS1基因及GSK-3β蛋白表达量均显著增加(P<0.01),穿越平台次数显著减少(P<0.01),游泳速度显著降低(P<0.01),超微结构损伤明显;与SAMP8组比较,安理申组与益肾化浊方组游泳总路程明显缩短(P<0.01),穿越平台次数明显增加(P<0.01),游泳速度亦显著加快(P<0.01),同时显著减轻神经元超微结构损伤;此外,安理申可显著下调GSK-3β蛋白表达(P<0.01),益肾化浊方可显著减少神经元凋亡数目(P<0.01),同时下调APP、PS1基因表达(P<0.01);组别和训练天数对逃避潜伏时间、总路程、穿越平台次数有明显影响(P<0.01)。结论益肾化浊方可有效提高SAMP8的空间记忆能力,抗神经元凋亡并保护神经元超微结构,其作用机制可能与下调APP、PS1基因和GSK-3β蛋白的表达相关。
Objective To study effects and mechanisms of Yishenhuazhuo recipe( YSHZR) on spatial learning and memory and the hippocampus neurons morphology in the senescence-accelerated prone 8( SAMP8) mice. Methods Forty-eight of 9-month-old SAMP8 mice were randomly allocated to model( SAMP8 mice),Aricept treated and YSHZR treated groups. Meanwhile,sixteen senescence-resistant 1( SAMR1) mice were used as controls( SAMR1 group).Aricept( 30 mg/kg),YSHZR( 20 g/kg) or equal-volume saline( SAMR1 and SAMP8mice) was administered in each mouse for 14 weeks by gastric infusion twice per day.Spatial learning and memory were measured by the Morris water maze.Neuronal apoptosis was quantified by Nissl staining.The mRNA expression of APP and PS1 and the protein expression of GSK-3β in the hippocampus were detected by RT-PCR and Western blot respectively.Results Compared with SAMR1 group,the path length,escape latency,number of neuronal apoptosis,GSK-3β protein and APP,PS1 gene expression in hippocampal CA1 of SAMP8 mice were significantly increased( P<0.01),frequency of entrance was significantly reduced( P < 0. 01),the swim speed was also significantly lowered( P <0.01),and the neuronal ultrastructural damage was very evident; compared with SAMP8 group,the path length in Aricept group and YSHZR group of mice were significantly shorter( P<0.01),the frequency of entrance was significantly increased( P<0.01),swim speed was also significantly faster( P<0.01),and neuronal ultrastructure damage was significantly reduced; moreover,Aricept significantly reduced GSK-3β protein expression( P<0.01),but YSHZR could significantly reduce the number of neuronal apoptosis( P<0.01),and down-regulate the gene expression of APP,PS1( P<0.01); groups and training days had significant effects on escape latency,total distance and the number of cross platform( P<0.01). Conclusions YSHZR plays a role of improving spatial memory,reducing neuronal apoptosis and protecting ultrastructure,which may be related to down-regulating the expression of APP,PS1 gene and GSK-3β protein.
引文
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