β片层阻断肽H102对双转基因AD小鼠脑内APP代谢分泌酶的影响

发布时间：2018-04-11 02:40

  本文选题：Aβ + APP代谢酶　； 参考：《天津医科大学》2017年硕士论文

【摘要】：目的:观察β片层阻断肽H102对APP/PS1双转基因AD小鼠脑内Aβ、APP代谢分泌酶及AD小鼠学习记忆能力的影响。方法:1.随机将30只6个月龄的APP/PS1双转基因小鼠分为AD组和H102组,同月龄和同背景的15只C57BL/6J小鼠作为对照组。采用鼻腔给药的方法,同一时间段,每天给药一次,H102组小鼠给予H102溶液(5.8 mg/kg)5μl,AD组和对照组给予辅料溶液5μl;连续给药30天后通过新物体识别实验和Morris水迷宫实验测试各组小鼠的学习记忆能力。2.行为学实验完成之后,10%的水合氯醛腹腔注射麻醉,PBS灌流、处死小鼠、冰上取脑,将一侧脑组织放于提前准备好的4%的多聚甲醛中,24h后进行包埋,将包埋好的组织制作成切片,用于免疫组织化学法检测脑内ADAM10、ADAM17、BACE1、PS1、PEN-2、APH1-a、Aβ蛋白的表达;另一半将海马和皮质分离后,做好标记先短暂保存在液氮中,然后再从液氮中取出放入-80℃冰箱保存,用于ELISA检测Aβ蛋白的表达和Western blot检测脑内ADAM10、ADAM17、BACE1、PS1、PEN-2、APH1-a蛋白的表达。结果:1.新物体识别实验:AD组相比对照组,新物体的识别指数明显降低(P0.05);H102组相比AD组,新物体的识别指数明显升高(P0.05);H102组与对照组相比,无显著性差异(P0.05)。2.定位航行实验:AD组相比对照组,逃避潜伏期延长(P0.05);H102组相比AD组,逃避潜伏期缩短(P0.05);然后进行空间探索实验,AD组相比对照组,第三象限停留时间和跨越隐匿平台的次数明显减少,入水朝向角明显增大(P0.05);H102组相比AD组,第三象限停留时间和跨越隐匿平台的次数明显增加,入水朝向角减小(P0.05);H102组与对照组相比,均无显著性差异(P0.05)。3.免疫组化测试:AD组小鼠脑内BACE1、PS1、PEN-2、APH1-a蛋白表达较对照组显著升高,ADAM10和ADAM17的蛋白表达相比于对照组显著降低(P0.05);H102组Aβ、BACE1、PS1、PEN-2、APH1-a蛋白比AD组显著降低,ADAM10、ADAM17蛋白表达比AD组显著升高(P0.05);H102组与对照组相比,均无显著性差异(P0.05)。4.Western blot检测:AD组小鼠脑内BACE1、PS1、PEN-2、APH1-a蛋白表达比对照组增高,ADAM10和ADAM17蛋白的表达比对照组减少(P0.05);H102组BACE1、PS1、PEN-2、APH1-a蛋白比AD组显著降低,ADAM10和ADAM17蛋白的表达比AD组显著升高增加(P0.05);H102组与对照组相比,均无显著性差异(P0.05)。5.Aβ蛋白检测结果:免疫组化和ELISA实验方法检测均表明,AD组相比对照组,Aβ蛋白的表达显著升高(P0.05);H102组相比AD组,Aβ蛋白的表达明显减少(P0.05);H102组与对照组相比,无显著性差异(P0.05)。结论:β片层阻断肽H102可影响AD小鼠脑内APP的代谢过程,减少Aβ、BACE1、PS1、PEN-2、APH1-a蛋白的表达,增加ADAM10、ADAM17蛋白的含量,减少Aβ的生成,H102可以增强AD小鼠的学习记忆能力。
[Abstract]:Aim: to observe the effects of 尾 -lamellar blocking peptide H102 on the metabolic secretory enzymes of A 尾 -app and the learning and memory ability of AD mice in APP/PS1 double transgenic AD mice.Method 1: 1.Thirty 6-month-old APP/PS1 double transgenic mice were randomly divided into AD group and H102 group. Fifteen C57BL/6J mice of the same age and the same background were used as control group.By nasal administration, at the same time,The mice in H102 group were given 5 渭 l excipient solution once a day with 5.8 mg/kg)5 渭 l AD and 5 渭 l in control group, and the learning and memory abilities of each group were tested by new object recognition test and Morris water maze test after 30 days of continuous administration.After the behavioral experiment was completed, 10% chloral hydrate was injected intraperitoneally into the anesthetized PBS, the mice were killed, the brain was taken from ice, one side of the brain tissue was embedded in 4% paraformaldehyde prepared in advance for 24 hours, and the embedded tissue was made into sections.Immunohistochemical method was used to detect the expression of ADAM10, ADAM17, BACE1, PS1, PEN-2, APH1-a 尾, and the other half, after separating hippocampus and cortex, stored in liquid nitrogen for a short time, then removed it from the liquid nitrogen and stored it in -80 鈩，

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