荭草苷对APP/PS1转基因痴呆小鼠认知功能和海马自噬的影响
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摘要
目的探究荭草苷对APP/PS1转基因小鼠认知功能的影响及其可能的作用机制。方法实验动物分为3组:7月龄的转基因模型组(Tg)、荭草苷处理转基因组(Tg+Ori)、同月龄的野生型C57BL/6J小鼠作为非转基因对照组(NT),每组8只。Tg+Ori组连续30 d每天腹腔注射荭草苷(10 mg·kg~(-1)),NT和Tg组注射同等剂量的生理盐水。水迷宫实验检测学习记忆能力,免疫组化检测β淀粉样蛋白(Aβ),免疫印迹检测自噬相关蛋白。结果与NT组相比,Tg组小鼠学习记忆能力受损,海马出现大量Aβ斑块, LC3-Ⅱ、p62、Cathepsin D蛋白表达升高;与Tg组相比,Tg+Ori组小鼠学习记忆能力增强,海马Aβ斑块减少,LC3-Ⅱ、p6、Cathepsin D蛋白水平降低;组间Beclin-1蛋白水平无明显差异。结论荭草苷能够改善转基因小鼠认知功能,减少海马Aβ沉积,促进自噬溶酶体降解。
Aim To study the influence of orientin on the cognitive function in APP/PS1 double transgenic mice and the probable mechanisms. Methods The mice(7 months of age) were randomly assigned into three groups(n=8 in each group): non-transgenic mice(NT) and APP/PS1 transgenic mice(Tg) were given saline, and APP/PS1 transgenic mice with orientin(Tg+Ori), and were injected intraperitoneally once per day for 30 days. Morris water maze test was carried out for the evaluation of spatial learning and memory. The measurement of Aβ was conducted by immunohistochemical staining. Autophagy related protein expressions of LC3-Ⅱ, p62, Cathepsin D and Beclin-1 were measured by Western blot. Results Compared with NT group, Tg mice showed deficit memory formation, enhanced Aβ deposition and protein levels of LC3-Ⅱ, p62 and Cathepsin D; compared with Tg group, Tg+Ori mice showed improved learning and memory, reduced Aβ load, and decreased levels of LC3-Ⅱ,p62 and Cathepsin D. The expression of Beclin-1 showed no difference between groups. Conclusions Orientin could improve the cognitive function, reduce amyloid plaques and ameliorate hippocampus autophagic clearance of APP/PS1 transgenic mice.
Aim To study the influence of orientin on the cognitive function in APP/PS1 double transgenic mice and the probable mechanisms. Methods The mice(7 months of age) were randomly assigned into three groups(n=8 in each group): non-transgenic mice(NT) and APP/PS1 transgenic mice(Tg) were given saline, and APP/PS1 transgenic mice with orientin(Tg+Ori), and were injected intraperitoneally once per day for 30 days. Morris water maze test was carried out for the evaluation of spatial learning and memory. The measurement of Aβ was conducted by immunohistochemical staining. Autophagy related protein expressions of LC3-Ⅱ, p62, Cathepsin D and Beclin-1 were measured by Western blot. Results Compared with NT group, Tg mice showed deficit memory formation, enhanced Aβ deposition and protein levels of LC3-Ⅱ, p62 and Cathepsin D; compared with Tg group, Tg+Ori mice showed improved learning and memory, reduced Aβ load, and decreased levels of LC3-Ⅱ,p62 and Cathepsin D. The expression of Beclin-1 showed no difference between groups. Conclusions Orientin could improve the cognitive function, reduce amyloid plaques and ameliorate hippocampus autophagic clearance of APP/PS1 transgenic mice.
引文
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[2] Menzies F M,Fleming A,Caricasole A,et al.Autophagy and neurodegeneration:pathogenic mechanisms and therapeutic opportunities[J].Neuron,2017,93(5):1015-34.
[3] Yu L,Wang S,Chen X,et al.Orientin alleviates cognitive deficits and oxidative stress in Aβ1-42-induced mouse model of Alzheimer?s disease[J].Life Sci,2015,121:104-9.
[4] Wang S,Yu Y,Feng Y,et al.Protective effect of the orientin on noise-induced cognitive impairments in mice[J].Behav Brain Res,2016,296:290-300.
[5] Vorhees C V,Williams M T.Morris water maze:procedures for assessing spatial and related forms of learning and memory[J].Nat Protoc,2006,1(2):848-58.
[6] Hardy J,Allsop D.Amyloid deposition as the central event in the aetiology of Alzheimer?s disease[J].Trends Pharmacol Sci,1991,12(10):383-8.
[7] Kurt M A,Davies D C,Kidd M,et al.Neurodegenerative changes associated with beta-amyloid deposition in the brains of mice carrying mutant amyloid precursor protein and mutant presenilin-1 transgenes[J].Exp Neurol,2001,171(1):59-71.
[8] Kurt M A,Davies D C,Kidd M,et al.Hyperphosphorylated tau and paired helical filament-like structures in the brains of mice carrying mutant amyloid precursor protein and mutant presenilin-1 transgenes[J].Neurobiol Dis,2003,14(1):89-97.
[9] 刘立亚,王慧晔,黄秀兰.基于细胞自噬的荭草苷抗心肌缺血/再灌注损伤的机制研究[J].中国药理学通报,2016,32(4):542-7.[9] Liu L Y,Wang H Y,Huang X L.Mechanism of anti-myocardial ischemia/reperfusion injury of orientin based on autophagy[J].Chin Pharmacol Bull,2016,32(4):542-7.
[10] Lam K Y,Ling A P,Koh R Y,et al.A review on medicinal properties of orientin[J].Adv Pharmacol Sci,2016,2016:4104595.
[11] Schaaf M B,Keulers T G,Vooijs M A,Rouschop K M.LC3/GABARAP family proteins:autophagy-(un)related functions[J].FASEB J,2016,30(12):3961-78.
[12] Bjrky G,Lamark T,Brech A,et al.p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death[J].J Cell Biol,2005,171(4):603-14.
[13] Bento C F,Renna M,Ghislat G,et al.Mammalian autophagy:how does it work [J]?Annu Rev Biochem,2016,85:685-713.
[14] Vidoni C,Follo C,Savino M,et al.The role of Cathepsin D in the pathogenesis of human neurodegenerative disorders[J].Med Res Rev,2016,36(5):845-70.
[15] Lee J K,Jin H K,Park M H,et al.Acid sphingomyelinase modulates the autophagic process by controlling lysosomal biogenesis in Alzheimer?s disease[J].J Exp Med,2014,211(8):1551-70.