京尼平苷酸对APP/PS1小鼠学习记忆能力及脑内淀粉样蛋白沉积的影响
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摘要
目的探讨京尼平苷酸(GPA)对阿尔茨海默病转基因小鼠学习记忆能力的影响及对脑内淀粉样蛋白沉积的作用及机制。方法 APP/PS1转基因小鼠随机分为转基因组、GPA 25,50,75 mg/kg组及多奈哌齐组。C57BL/6小鼠为野生组。灌胃给药90 d后,通过Morris水迷宫实验评价痴呆小鼠的学习记忆能力。硫黄素-T染色检测小鼠脑组织内硫黄素-T阳性斑块的沉积。Western Blot法检测小鼠大脑皮层及海马内BACE及IDE蛋白水平。ELISA法检测小鼠大脑皮层及海马内IL-1β,IL-6,IL-4,TNF-α的水平。结果与野生组比较,转基因小鼠潜伏期明显增加,穿越平台次数明显减少(P<0. 05),脑内Th-T斑块百分面积显著增多(P<0. 01),IDE的蛋白水平显著降低且BACE的蛋白水平显著升高(P<0. 01),IL-1β,IL-6和TNF-α含量均显著升高,IL-4含量显著降低(P<0. 01)。与转基因小鼠比较,GPA 75 mg/kg剂量组转基因小鼠潜伏期明显缩短,穿越平台次数明显增多(P<0. 05),GPA处理组小鼠脑内Th-T荧光面积百分比显著减少,IDE水平显著升高,BACE水平显著降低(P<0. 01),GPA 50,75 mg/kg组小鼠脑内IL-1β,IL-6和TNF-α含量均显著降低(P<0. 01或P<0. 05),GPA 50 mg/kg明显升高小鼠皮层IL-4含量(P<0. 05)。结论京尼平苷酸可通过影响Aβ的生成和降解两方面调控小鼠脑内Aβ水平,减轻转基因小鼠的神经炎症,发挥神经保护作用。
Objective To investigate the effect and protect mechanisms of geniposidic acid on learning and memory defects and fibrillarβ-amyloid aggregations in APP/PS1 mice. Method Double transgenic APP/PS1 mice were randomly assigned into transgenic group,GPA 25 mg/kg group,GPA 50 mg/kg group,GPA 75 mg/kg group and donepezil group. C57 BL/6 J mice were used as wide-type group. Treatments were administered by oral gavage daily for ninety days. Morris water maze was tested to estimate the memory and spatial learning abilities of mice. Thioflavin-T( Th-T) staining were applied to assess fibrillar β-amyloid aggregations in brain. The expression of BACE and IDE in the cortex and the hippocampus of mice were detected by Western blot. In order to quantify the expression of IL-1β,IL-6,IL-4,and TNF-α,ELISA kits were applied. Results Compared with wide-type mice,transgenic mice exhibited shorter escape latency and a reduced number of crossings( P<0. 05),the percent area of Th-T-positive plaque in the cortex and hippocampus increased significantly( P<0. 01),the protein expression of BACE increased and IDE decreased significantly( P<0. 01),the expression of IL-1β,IL-6 and TNF-α reduced and IL-4 induced in brain significantly( P< 0. 01). Compared with transgenic mice,mice in 75 mg/kg GPA treatment groups had shortened escape latency and improved target searching capacity markedly( P< 0. 05).GPA could reduce the percent area of Th-T-positive plaque and the protein expression of BACE,increase the protein expression of IDE in brain of mice( P<0. 01). 50,75 mg/kg GPA could obviously increase the expression of IL-4 in the cortex( P<0. 05) and decrease the expression of IL-1β,IL-6 and TNF-α significantly in brain( P<0. 01 or P<0. 05). Conclusion GPA could effectively regulate the level of Aβ in brain by effecting the production and degradation of Aβ,reducing neuroinflammation of brain,therefore protecting the neurons.
Objective To investigate the effect and protect mechanisms of geniposidic acid on learning and memory defects and fibrillarβ-amyloid aggregations in APP/PS1 mice. Method Double transgenic APP/PS1 mice were randomly assigned into transgenic group,GPA 25 mg/kg group,GPA 50 mg/kg group,GPA 75 mg/kg group and donepezil group. C57 BL/6 J mice were used as wide-type group. Treatments were administered by oral gavage daily for ninety days. Morris water maze was tested to estimate the memory and spatial learning abilities of mice. Thioflavin-T( Th-T) staining were applied to assess fibrillar β-amyloid aggregations in brain. The expression of BACE and IDE in the cortex and the hippocampus of mice were detected by Western blot. In order to quantify the expression of IL-1β,IL-6,IL-4,and TNF-α,ELISA kits were applied. Results Compared with wide-type mice,transgenic mice exhibited shorter escape latency and a reduced number of crossings( P<0. 05),the percent area of Th-T-positive plaque in the cortex and hippocampus increased significantly( P<0. 01),the protein expression of BACE increased and IDE decreased significantly( P<0. 01),the expression of IL-1β,IL-6 and TNF-α reduced and IL-4 induced in brain significantly( P< 0. 01). Compared with transgenic mice,mice in 75 mg/kg GPA treatment groups had shortened escape latency and improved target searching capacity markedly( P< 0. 05).GPA could reduce the percent area of Th-T-positive plaque and the protein expression of BACE,increase the protein expression of IDE in brain of mice( P<0. 01). 50,75 mg/kg GPA could obviously increase the expression of IL-4 in the cortex( P<0. 05) and decrease the expression of IL-1β,IL-6 and TNF-α significantly in brain( P<0. 01 or P<0. 05). Conclusion GPA could effectively regulate the level of Aβ in brain by effecting the production and degradation of Aβ,reducing neuroinflammation of brain,therefore protecting the neurons.
引文
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[2]彭也,朱天碧,张钊,等.阿尔茨海默病的免疫调控研究进展[J].中国药理学通报,2019(4):445-451.Peng Y,Zhu T B,Zhang Z,et al.The research progress of immune regulation of Alzheimer’s disease[J].Chinese Pharmacological Bulletin,2019(4):445-451.
[3]Arbor S C,Lafontaine M,Cumbay M.Amyloid-beta alzheimer targets protein processing,lipid rafts,and amyloidbeta pores[J].The Yale Journal of Biology and Medicine,2016,89(1):5-21.
[4]Uchoa M F,Alexandra Moser V,Pike C J.Interactions between inflammation,sex steroids,and Alzheimer’s disease risk factors[J].Frontiers in Neuroendocrinology,2016,43:60-82.
[5]Kwon S H,Lee H K,Kim J A,et al.Neuroprotective effects of eucommia ulmoides oliv bark on amyloid beta 25!35-induced learning and memory impairments in mice[J].Neuroscience Letters,2001,487(1):123-127.
[6]Zhou Y Q,Liang M,Li W Z,et al.Protective effects of eucommia ulmoides oliv bark and leaf on amyloidβ-induced cytotoxicity[J].Environ Toxicol Pharmacol,2009,28(3):342-349.
[7]Kwon S H,Ma S X,Hwang J Y,et al.The anti-inflammatory activity of eucommia ulmoides oliv bark involves NF-κB suppression and Nrf2-dependent HO-1 induction in BV-2 microglial cells[J].Biomolecules&Therapeutics,2016,24(3):268-282.
[8]Kwon S H,Kim M J,Ma S X,et al.Eucommia ulmoides oliv bark protects against hydrogen peroxide-induced neuronal cell death in SH-SY5Y cells[J].Journal of Ethnopharmacology,2012,142(2):337-345.
[9]周张玖智,丁杨芳,胡艳丽.京尼平苷酸对D-半乳糖/亚硝酸钠诱导的阿尔茨海默病小鼠学习记忆的影响[J].中国老年学杂志,2019,39(5):1188-1191.Zhou Z J Z,Ding Y F,Hu Y L.Effect of geniposidic acid on D-galactose/sodium nitrite induced learning and memory disorders in mice[J].Chinese Journal of Gerontology,2019,39(5):1188-1191.
[10]Huang Y W A,Zhou B,Wernig M,et al.Apo E2,Apo E3,and Apo E4 differentially stimulate APP transcription and Aβsecretion[J].Cell,2017,168(3):427-441
[11]Kanekiyo T,Xu H,Bu G.Apo E and Aβin alzheimer’s disease:accidental encounters or partners[J].Neuron,2014,81(4):740-754.
[12]Song C,Zhang Y,Dong Y.Acute and subacute IL-1βadministrations differentially modulate neuroimmune and neurotrophic systems:possible implications for neuroprotection and neurodegeneration[J].Journal of Neuroinflammation,2013,10(1):59.
[13]崔勇,王艳杰,赵丹玉,等.六味地黄丸对APP/PS1小鼠学习记忆能力及胆碱能系统的影响[J].中国实验方剂学杂志,2016,22(3):148-151.Cui Y,Wang Y J,Zhao D Y,et al.Effects of liuwei dihuang wan on learning and memory abilities and cholinergic system in APP/PS1 mice[J].Chinese Journal of Experimental Traditional Medical Formulae,2016,22(3):148-151.
[14]Lindberg D J,Wenger A,Sundin E,et al.Binding of thioflavin-T to amyloid fibrils leads to fluorescence Selfquenching and fibril compaction[J].Biochemistry,2017,56(16):2170-2174.
[15]蔺莹,姚璎珈,梁喜才,等.蛇床子素上调miRNA-9抑制BACE-1表达治疗阿尔茨海默病[J].中国药理学通报,2019(4):524-529.Lin Y,Yao Y J,Liang X C,et al.Osthole suppresses BACE-1 expression by up-regulating miR-9 in Alzheimer’s disease[J].Chinese Pharmacological Bulletin,2019(4):524-529.
[16]Heneka M T,Sastre M,Dumitrescu-Ozimek L,et al.Focal glial activation coincides with increased BACE1 activation and precedes amyloid plaque deposition in APP transgenic mice[J].Journal of Neuroinflammation,2005,2(1):22.
[17]张爽曦,龙建纲,刘健康.胰岛素降解酶与阿尔兹海默病的发生[J].中国生物化学与分子生物学报,2018,34(3):249-254.Zhang S X,Long J G,Liu J K.The Role of the insulin degrading enzyme in the pathogenesis of alzheimer's disease Chinese[J].Journal of Biochemistry and Molecular Biology,2018,34(3):249-254.
[18]Son S M,Cha M Y,Choi H,et al.Insulin-degrading enzyme secretion from astrocytes is mediated by an autophagy-based unconventional secretory pathway in Alzheimer disease[J].Autophagy,2016,12(5):784-800.
[19]Farris W,Mansourian S,Leissring M A,et al.Partial loss-of-function mutations in insulin-degrading enzyme that induce diabetes also impair degradation of amyloidβ-Protein[J].American Journal of Pathology,2004,164(4):1425-1434.
[20]Farris W,Mansourian S,Chang Y,et al.Insulin-degrading enzyme regulates the levels of insulin,amyloidβ-protein,and theβ-amyloid precursor protein intracellular domain in vivo[J].Proceedings of the National Academy of Sciences,2003,100(7):4162-4167.