盐酸多奈哌齐对帕金森大鼠认知功能及大脑海马组织Klotho、Akt和Foxo3a表达的影响

Effects of donepezil hydrochloride on cognitive function and expression of Klotho, Akt and Foxo3a in hippocampus of rats model of Parkinson disease

目的探讨盐酸多奈哌齐对于6-羟基多巴胺(6-OHDA)致帕金森大鼠认知功能及海马组织Klotho、Akt和Foxo3a mRNA及蛋白的影响。方法选取同批次饲养健康雄性SD大鼠,分为对照组(同剂量生理盐水)、帕金森疾病模型组(4μg/μl 6-OHDA+同剂量生理盐水)、盐酸多奈哌齐低剂量组(4μg/μl 6-OHDA+3 mg/kg·bw盐酸多奈哌齐)和盐酸多奈哌齐高剂量组(4μg/μl 6-OHDA+6 mg/kg·bw盐酸多奈哌齐),使用6-OHDA制作帕金森病模型大鼠,利用Morris水迷宫实验评价大鼠认知功能,苏木素-伊红(HE)染色观察大鼠海马区病理变化,ELISA法检测各组大鼠血清中SOD和MDA含量,用Western blot法检测海马区Klotho、p-Akt和p-Foxo3a蛋白的表达水平,荧光定量PCR检测大鼠脑海马区Klotho、Akt和Foxo3a mRNA表达。结果帕金森大鼠神经元细胞排列疏松、紊乱,细胞核固缩、深染,细胞质减少。高剂量组大鼠水迷宫试验完成时间明显低于模型组和低剂量组大鼠(P<0.05),海马区Klotho蛋白及mRNA表达高于模型组和低剂量组大鼠(P<0.05),p-Akt和p-Foxo3a蛋白表达明显低于模型组和低剂量组(P<0.05),Akt和Foxo3a mRNA表达高于模型组和低剂量组(P<0.05),血清SOD表达高于模型组和低剂量组(P<0.05),MDA表达低于模型组和低剂量组(P<0.05),差异均有统计学意义。结论盐酸多奈哌齐可能通过上调海马区Klotho mRNA表达,抑制Akt和Foxo3a磷酸化,提高血清SOD含量,在一定程度上改善帕金森模型大鼠的学习记忆能力。

Objective To investigate the mechanism of donepezil hydrochloride on the aging related genes Klotho,Akt and Foxo3a in the hippocampus of Parkinson model rats.Methods Model rats with Parkinson disease were established.Sixty SD rats were divided into the control group,the model group,the saline group and the donepezil hydrochloride group with 10 rats in each group.The Morris water maze test was used to evaluate the cognitive impairment in rats.ELISA was used to detect the contents of SOD and MDA in serum of each group,The expression level of Klotho,p-Akt,p-Foxo3a protein in hippocampal region by Western blot and the expression of Klotho,Akt and Foxo3a mRNA in hippocampus of rats was detected by quantitative PCR.Results The arrangement of neurons in rats of Parkinson disease was loose and disordered,the nucleus of neurons was pyknosis and hyperchromatic,and the cytoplasm was reduced.The time of water maze test in high dose group was significantly lower than that of model group and low dose group(P<0.05).The expression of Klotho protein and mRNA in hippocampus was significantly higher than that of model group and low dose group(P<0.05).The expression of p-Akt and p-Foxo3a protein was significantly lower than that of model group and low dose group(P<0.05),Akt and Foxo3a mRNA expression significantly higher than model group and low dose group(P<0.05),the expression of serum SOD was significantly higher than that of model group and low dose group(P<0.05),and the expression of MDA was significantly lower than that of model group and low dose group(P<0.05).Conclusion Donepezil can increase the expression of Klotho mRNA in the hippocampus,inhibit the phosphorylation of Akt and Foxo3a,increase the content of serum SOD,so as to improve the learning and memory ability of rats Parkinson model.