补肾法中药调控ERK/MAPK信号通路对Aβ25-35诱导的阿尔茨海默病大鼠模型的神经保护作用研究

Neuroprotective Study of Tonifying Kidney TCM Drugs in Aβ25-35-induced Alzheimer′s Disease in Rat Model by Regulating ERK/MAPK Signaling Pathway

目的 观察补肾法中药通过调控ERK/MAPK信号通路关键靶点蛋白,探讨其对Aβ25-35诱导的阿尔茨海默病(AD)大鼠模型的神经保护作用机制。方法 取36只雌性SD大鼠随机分为假手术组、模型组、中药组,每组12只。采用Aβ25-35侧脑室注入建立AD大鼠模型,中药组予补肾中药,模型组及假手术组予蒸馏水灌胃,观察各组大鼠认知记忆能力、海马神经元凋亡、p-ERK1/2蛋白表达情况。结果 1)AD认知记忆功能检测:与模型组相比,中药组大鼠定位航行实验显示,平均速度无显著差异(P>0.05),潜伏时间明显缩短,学习能力明显上升,差异具有统计学意义(P<0.05)。2)AD海马神经元凋亡检测:光镜下观察尼氏染色结果显示,与模型组相比,中药组神经元凋亡数量明显减少(P<0.01)。3)ERK/MAPK信号通路关键靶点p-ERK1/2蛋白表达的检测:Western blotting检测结果显示,与假手术组相比,模型组p-ERK1/2蛋白的表达均明显降低,差异具有统计学意义(P<0.05);与模型组相比,中药组p-ERK1/2蛋白的表达明显增加,差异具有统计学意义(P<0.05)。结论 补肾法中药对Aβ25-35诱导的AD模型大鼠海马神经元损伤具有保护作用,可明显改善AD模型大鼠认知和记忆能力,减少海马神经元的凋亡,其作用机制可能调控p-ERK1/2蛋白表达,激活ERK/MAPK信号通路有关。

Objective:To observe the influence of Tonifying kidney TCM drugs on the key target proteins of ERK/MAPK signaling pathway,and to explore the neuroprotective mechanism of Tonifying kidney TCM drugs in Aβ25-35 induced Alzheimer′s disease(AD)rat model.Methods:36 SD female rats were randomly divided into the sham AD group,the AD modle group(group AD),and the Bushen AD group(group AD+BS).12 rats were in each group.AD model was established by Aβ25-35 paracele injection,and the changes of cognitive memory func?tion and hippocampal neuron apoptosis in the AD model were observed from macro to the microscopic level,and the changes in the expression of p-ERK1/2 protein in the key target of the ERK/MAPK signaling pathway were ob?served.Results:1)Cognitive and memory function test:Compared with group AD,the group AD+BS showed that there was no significant difference in average speed(P>0.05),but latency time was significantly shortened,and learning ability was significantly increased,and the differences were statistically significant(P<0.05).2)Detection of hippocampal neuronal apoptosis:Nissl staining results showed that compared with group AD,the number of neu?ron apoptosis in group AD+BS was significantly reduced(P<0.01).3)The expression of p-ERK1/2 protein,the key target protein of ERK/MAPK signaling pathway,was detected by Western blotting.Compared with group Sham AD,the expression of p-ERK1/2 protein in group AD and group AD+BS was significantly decreased(P<0.05).Compared with group AD,the expression of p-ERK1/2 protein in group AD+BS was significantly increased,and the difference was statistically significant(P<0.05).Conclusion:Tonifying kidney TCM drugs has a protective effect on hippocampal neuron injury induced by Aβ25-35 in AD model rats,which can significantly improve cogni?tion and memory ability,and reduce hippocampal neuron apoptosis in AD model rats.The mechanism may be relat?ed to regulating p-ERK1/2 protein expression and activating ERK/MAPK signaling pathway.