艾司西酞普兰对MPTP诱导急性帕金森小鼠模型的运动和认知功能的影响

发布时间：2018-05-18 21:06

本文选题：艾司西酞普兰 + 帕金森病　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:帕金森病(Parkinson's disease,PD)是常发于中老年人的中枢神经系统变性疾病,主要临床表现为震颤、强直、运动弛缓、姿势平衡障碍等运动症状,此外,常伴发抑郁、焦虑、便秘、睡眠障碍、认知功能减退等非运动症状。艾司西酞普兰(escitalopram,ESC)是新型的高选择性5-羟色胺再摄取抑制剂(Selective Serotonin Reuptake Inhibitor,SSRI),对受体的选择性较高,对肝药酶P450的抑制作用小,不良反应及药物相关反应也较少,安全性较高,在临床广泛应用于广泛性焦虑、抑郁症。本文旨在探讨ESC对PD小鼠模型运动、认知功能的影响。方法:将37只11周龄23~30g C57BL/6J健康雄性小鼠随机分为3组:1正常对照组(CON组):正常健康小鼠,腹腔注射0.1ml生理盐水;2急性PD组(PD组):采用急性腹腔注射MPTP制作急性PD模型小鼠,20mg/kg MPTP腹腔注射,每2h注射1次,共4次;3 ESC预处理组:在制备PD模型前3天,每日对小鼠进行10mg/kg ESC腹腔注射,每天1次,共3天;以MPTP制备PD模型为第0天,第-3至-1天进行爬杆实验适应性训练及ESC预处理,第2-7天进行Morris水迷宫实验,第8天进行爬杆实验,第9天按照预定条件取各组小鼠大脑,采用Western blot观察小鼠大脑海马BDNF的变化,采用免疫组织化学观察小鼠海马细胞个数变化,以及观察黑质-纹状体酪氨酸羟化酶阳性(tyrosine hydroxylase positive,TH+)细胞的改变。结果:1黑质及纹状体TH+细胞的改变第15d小鼠黑质行TH免疫组织化学染色,结果显示:PD组小鼠黑质的残留TH+细胞皱缩,不饱满,有梭形改变,突起减少。PD组与CON组相比,黑质TH+细胞数较正常下降49.31%,ESC预处理组与CON组相比,黑质TH+细胞数较正常下降32.35%。差异均有统计学意义(P0.05)。PD组与CON组相比,PD组纹状体TH+细胞的平均光密度(mean optical density,MOD)较CON组下降了69.55%,ESC预处理组较CON组下降了51.46%,差异有统计学意义(P0.05)。2 ESC对PD模型小鼠认知功能影响2.1 ESC对PD模型小鼠学习功能的影响PD模型造模成功后,在第2天进行Morris水迷宫实验测试各组小鼠的学习功能。通过对定向航行实验数据进行重复测量检验,各组小鼠的潜伏期均逐渐缩短(CON组=ESC组PD组),CON组与PD组差异有统计学意义(P0.05),CON组与ESC预处理组差异无统计学意义(P0.05),PD组与ESC组差异有统计学意义(P0.05),CON组和ESC组较PD组潜伏期明显缩短。2.2 ESC对PD模型小鼠记忆功能的影响通过第6天水迷宫检测,CON组小鼠与PD组小鼠穿过平台次数及穿越平台路程比率差异有统计学意义(P0.05),CON组与ESC预处理组差异无统计学意义(P0.05),ESC预处理组有穿越平台次数增加及穿越平台路程比率升高但与PD组差异无统计学意义(P0.05)。小鼠的记忆状况为CON组≥ESC组≥PD组,且CON组PD组,说明PD小鼠的记忆能力明显下降,ESC预处理组的记忆能力在一定程度上有所改善,但是尚不能与PD组有明显区别。3 ESC对PD模型小鼠海马区BDNF含量的影响通过Western blot检测各组小鼠海马区BDNF含量,结果显示PD组(0.09±0.01)BDNF含量明显低于CON组(0.64±0.27)及ESC预处理组(0.57±0.05),差异有统计学意义(P0.05),而CON组与ESC预处理组无差异(P0.05)。4 ESC对PD模型小鼠海马区细胞个数的影响小鼠海马区细胞经免疫组化胆碱能染色后,结果显示CON组细胞数(100.9±10.8)个与PD组小鼠(84±7.0)个差异有统计学意义(P0.05),CON组与ESC预处理组(94.8±7.6)个差异无统计学意义(P0.05),PD组与ESC预处理组差异有统计学意义(P0.05),CON组和ESC组平均海马细胞个数明显高于PD组。5 ESC对PD模型小鼠运动功能的影响在第7d进行的Morris水迷宫试验检测各组小鼠游泳速度,结果显示:PD组与CON组、ESC预处理组相比游泳速度显著降低,差异有统计学意义(P0.05),CON组与ESC预处理组间的游泳速度相比,差异无统计学意义(P0.05)。在MPTP造模第8d进行爬杆实验,检测各组小鼠的运动功能。结果显示:CON组与PD组差异有统计学意义(P0.05),CON组与ESC预处理组差异无统计学意义(P0.05),PD组与ESC组有差异有统计学意义(P0.05),CON组与ESC预处理组小鼠的爬杆时间明显低于PD组。结论:1急性腹腔注射MPTP可以成功制作PD模型,在PD模型制作初期运动能力损伤严重,PD小鼠的震颤、强直症状明显,24h后,症状逐渐减轻至消失。MPTP对C57BL/6J小鼠的黑质-纹状体多巴胺能神经元和神经纤维造成了严重损害。2 MPTP对C57BL/6J小鼠的学习及记忆能力有损害,认知功能明显下降。ESC可以改善急性PD小鼠的学习能力,但对MPTP造成的记忆损害无明显改善;ESC预处理可以缓解急性PD小鼠的运动功能障碍。3 ESC预处理对PD小鼠的保护作用可能与ESC能够保护海马和黑质多巴胺神经元以及提高海马区BDNF有关。
[Abstract]:Objective: Parkinson's disease (Parkinson's disease, PD) is a central nervous system degenerative disease often occurring in middle-aged and old people. The main clinical manifestations are tremor, ankylosis, movement relaxation, postural balance disorder and other sports symptoms. In addition, it often accompanied by depression, anxiety, constipation, sleep barrier, cognitive impairment and other non motor symptoms. (escitalopr (escitalopr) Am, ESC) is a new high selective 5- serotonin reuptake inhibitor (Selective Serotonin Reuptake Inhibitor, SSRI), which has higher selectivity to the receptor, less inhibition to the liver enzyme P450, less adverse reactions and drug related reactions and high safety. It is widely used in widespread anxiety and depression in the bed. This paper aims to explore ESC. The influence of cognitive function on PD mouse model movement. Methods: 37 11 weeks old 23~30g C57BL/6J healthy male mice were randomly divided into 3 groups: 1 normal control group (group CON): normal healthy mice, intraperitoneal injection of 0.1ml physiological saline; 2 acute PD group (group PD): acute peritoneal injection of MPTP to make acute PD model mice, 20mg/kg MPTP intraperitoneal injection, each 2H 1 times, a total of 4 times; 3 ESC pretreatment group: 3 days before the preparation of the PD model, the mice were intraperitoneally injected with 10mg/kg ESC daily, 1 times a day, for a total of 3 days; MPTP was used to prepare PD model for zeroth days, -3 to -1 days for climbing pole experiment adaption training and ESC preprocessing, the 2-7 day Morris water maze experiment, eighth days climbing pole experiment, Ninth days according to preconditioning. The changes of BDNF in the hippocampus of mice were observed by Western blot. The changes in the number of hippocampal cells in mice were observed by immunohistochemistry and the changes of tyrosine hydroxylase positive (TH+) cells were observed in the substantia nigra and striatum (tyrosine hydroxylase positive, TH+). Results: the changes of 1 substantia nigra and striatum TH+ cells were changed by first 5D murine substantia nigra was stained with TH immunohistochemical staining. The results showed that the residual TH+ cells in the substantia nigra of the PD group were crinkled, not full, and there was a change of spindle shape. The number of TH+ cells in the substantia nigra decreased by 49.31% compared with the CON group in the.PD group, and the number of TH+ cells in the ESC preconditioning group compared with the CON group was statistically significant (P) was significantly lower than that of the normal 32.35%. (P). 0.05) compared with the CON group, the average light density (mean optical density, MOD) of the PD group was 69.55% lower than the CON group in the group PD, and the ESC pretreatment group was 51.46% lower than the CON group. The difference was statistically significant (P0.05) influenced the cognitive function of the model mice 2.1. The Morris water maze test was used to test the learning function of the mice in the Morris water maze test. Through the repeated measurement of the directional navigation test data, the incubation period of the mice was gradually shortened (group PD of group CON =ESC). The difference between the CON group and the PD group was statistically significant (P0.05), and there was no significant difference between the CON group and the ESC pretreatment group (P0.05), PD group. The difference between group ESC and group ESC was statistically significant (P0.05). The incubation period of group CON and ESC significantly shortened the effect of.2.2 ESC on the memory function of PD model mice through sixth days water maze, and there were statistical significance (P0.05) in the number of the CON group and the PD group through the platform times and the crossing platform, and there was no difference between the CON group and the pretreatment group. The study significance (P0.05), ESC pretreated group increased the number of crossing platform and the increase of cross platform road ratio, but there was no significant difference with the PD group (P0.05). The memory status of the mice was CON group > ESC group > PD group, and CON group PD group, indicating that the memory ability of PD mice descended obviously, and the memory ability of ESC pretreatment group was certain to some extent. However, the effect of.3 ESC on the BDNF content in hippocampus of PD model mice was not significantly different from that of PD group. The content of BDNF in the hippocampus of each group was detected by Western blot. The results showed that the content of PD group (0.09 + 0.01) BDNF was significantly lower than that of CON group (0.64 + 0.27) and ESC preconditioning group (0.57 + 0.05), and the difference was statistically significant. The effect of pre treated group (P0.05).4 ESC on the number of cells in the hippocampus of PD model mice was affected by immunohistochemical cholinergic staining, the results showed that the number of cells in the CON group (100.9 + 10.8) was significantly different from that of the PD group (84 + 7) (P0.05), and there was no significant difference between the CON group and the ESC preconditioning group (94.8 + 7.6). (P0.05), the difference between group PD and ESC preconditioning group was statistically significant (P0.05). The average number of hippocampal cells in group CON and ESC was significantly higher than that of PD group.5 ESC on the movement function of PD model mice. The swimming speed of mice in 7d Morris water maze test was detected in 7d Morris water maze test. The difference was statistically significant (P0.05). There was no significant difference in swimming speed between group CON and ESC preconditioning group (P0.05). The exercise function of mice in each group was detected in MPTP model 8D. The results showed that there was significant difference between group CON and PD group (P0.05), and there was no significant difference between the CON group and the ESC pretreatment group (P0.05). P0.05), there was a significant difference between group PD and group ESC (P0.05). The time of climbing pole of mice in group CON and ESC preconditioning group was obviously lower than that of group PD. Conclusion: 1 acute peritoneal injection of MPTP can make PD model successfully. In the early stage of making PD model, the exercise ability is seriously damaged, PD mice tremble, and the symptoms of ankylosis are obvious. After 24h, the symptoms gradually lighten to disappear. P had serious damage to the nigrostriatal dopaminergic neurons and nerve fibers in C57BL/6J mice..2 MPTP was impaired in the learning and memory ability of C57BL/6J mice. The cognitive decline of.ESC could improve the learning ability of acute PD mice, but the memory damage caused by MPTP was not significantly improved; ESC preconditioning could alleviate the acute toxicity of C57BL/6J. The protective effect of.3 ESC preconditioning in PD mice on PD mice may be related to the protective effect of ESC on the protection of hippocampus and substantia nigra dopamine neurons and the enhancement of BDNF in the hippocampus.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.5;R-332

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