CYP46A1过表达对阿尔茨海默病转基因模型小鼠的认知改善和抗炎症作用
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摘要
目的探讨细胞色素P450 46A1(CYP46A1)对阿尔茨海默病的影响。方法构建含人源CYP46A1的慢病毒载体,通过立体定位仪注射到3月龄的雄性5XFAD转基因小鼠双侧海马中,对照组在相应部位注射空载体。2个月后,通过Morris水迷宫和T迷宫实验检测小鼠的学习记忆能力,通过免疫组织化学染色和ELISA检测脑组织内β-淀粉样蛋白(Aβ)沉积情况和炎症水平。结果与对照组相比,CYP46A1过表达组小鼠海马中CYP46A1 mRNA和蛋白表达显著增加,学习记忆能力显著提高。小鼠脑组织海马区域的Aβ斑块面积和星形胶质细胞数量显著减少,β淀粉样蛋白40(Aβ_(40))、Aβ_(42)、白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)表达水平均显著下降。结论 CYP46A1过表达可改善5XFAD转基因小鼠的认知障碍及脑内炎症水平,提示CYP46A1具有抗阿尔茨海默病样的作用。
Objective To investigate the effect of CYP46A1 on the pathogenesis of Alzheimer 's disease. MethodsRecombinant lentiviral vectors which including anthropogenic CYP46A1 were injected into bilateral hippocampus of 3-monthold male 5 XFAD transgenic mice, while empty vectors were injected into the corresponding position of the control group. After two months, the ability of learning and memory were tested by Morris water maze and T maze experiments, and amyloid plaque and inflammator y infiltration in the brain were detected by immunohistochemical staining and ELISA respectively. ResultsCompared with the control group, CYP46A1 virus injection significantly increased the CYP46A1 mRNA and protein expression in hippocampus. In addition, CYP46A1 overexpression significantly decreased the latency to find the platform in Morris water maze test and increased the correct rate to choose in T maze test. Aβ immunohistochemical staining and plaques area statistics demonstrated that the amyloid plaque area of hippocampus in CYP46A1 overexpression mice was significantly reduced, and there was a significantly decrease of hippocampal astrocytes expression by means of GFAP staining. Furthermore, hippocampal CYP46A1 overexpression significantly decreased the expression level of Aβ_(40), Aβ_(42), IL-1β and TNF-α, while compare with the control group. Conclusion CYP46A1 overexpression in hippocampus can promote the cognitive impairment, as well as ameliorate the brain inflammatory infiltration in 5 XFAD transgenic mice, suggesting that CYP46A1 has anti-Alzheimer's disease like effects.
Objective To investigate the effect of CYP46A1 on the pathogenesis of Alzheimer 's disease. MethodsRecombinant lentiviral vectors which including anthropogenic CYP46A1 were injected into bilateral hippocampus of 3-monthold male 5 XFAD transgenic mice, while empty vectors were injected into the corresponding position of the control group. After two months, the ability of learning and memory were tested by Morris water maze and T maze experiments, and amyloid plaque and inflammator y infiltration in the brain were detected by immunohistochemical staining and ELISA respectively. ResultsCompared with the control group, CYP46A1 virus injection significantly increased the CYP46A1 mRNA and protein expression in hippocampus. In addition, CYP46A1 overexpression significantly decreased the latency to find the platform in Morris water maze test and increased the correct rate to choose in T maze test. Aβ immunohistochemical staining and plaques area statistics demonstrated that the amyloid plaque area of hippocampus in CYP46A1 overexpression mice was significantly reduced, and there was a significantly decrease of hippocampal astrocytes expression by means of GFAP staining. Furthermore, hippocampal CYP46A1 overexpression significantly decreased the expression level of Aβ_(40), Aβ_(42), IL-1β and TNF-α, while compare with the control group. Conclusion CYP46A1 overexpression in hippocampus can promote the cognitive impairment, as well as ameliorate the brain inflammatory infiltration in 5 XFAD transgenic mice, suggesting that CYP46A1 has anti-Alzheimer's disease like effects.
引文
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[20]Liu JJ,Cao N,Di JL,et al.Role of bone marrow mesenchymal stem ceils in repair of neural cell injury induced by okadaic acid[J].Med J Chin PLA,2017,42(5):377-382.[刘嘉婧,曹宁,翟晶磊,等.骨髓间充质干细胞对冈田酸致神经细胞损伤的修复作用[J].解放军医学杂志,2017,42(5):377-382.]
[2]Allinquant B,Clamagirand C,Potier MC.Role of cholesterol metabolism in the pathogenesis of Alzheimer's disease[J].Curr Opin Clin Nutr,2014,17(4):319-323.
[3]Maulik M,Westaway D,Jhamandas JH,et al.Role of cholesterol in APP metabolism and its significance in Alzheimer's disease pathogenesis[J].Mol Neurobiol,2013,47(1):37-63.
[4]Papassotiropoulos A,Lutjohann D,Bagli M,et al.Plasma24S-hydroxycholesterol:a peripheral indicator of neuronal degeneration and potential state marker for Alzheimer's disease[J].Neuroreport,2000,11(9):1959-1962.
[5]Mast N,Charvet C,Pikuleva IA,et al.Structural basis of drug binding to CYP46A1,an enzyme that controls cholesterol turnover in the brain[J].J Biol Chem,2010,285(41):31783-31795.
[6]Russell DW,Halford RW,Ramirez DM,et al.Cholesterol24-hydroxylase:an enzyme of cholesterol turnover in the brain[J].Annu Rev Biochem,2009,78:1017-1040.
[7]Garcia AN,Muniz MT,Souza e Si lva HR,et al.Cyp46polymorphisms in Alzheimer's disease:a review[J].J Mol Neurosci,2009,39(3):342-345.
[8]Djelti F,Braudeau J,Hudry E,et al.CYP46A1 inhibition,brain cholesterol accumulation and neurodegeneration pave the way for Alzheimer's disease[J].Brain,2015,138(Pt 8):2383-2398.
[9]Oakley H,Cole SL,Logan S,et al.Intra-neuronal beta-amyloid aggregates,neurodegeneration,and neuron loss in transgenic mice with five familial Alzheimer's disease mutations:potential factors in amyloid plaque formation[J].J Mol Neurosci,2006,26(40):10129-10140.
[10]Yan HC,Qu HD,Sun LR,et al.Fuzi polysaccharide-1produces ant idepressant-like ef fects in mice[J].Int JNeuropsychopharmacol,2010,13(5):623-633.
[11]Vorhees CV,Williams MT.Morris water maze:procedures for assessing spatial and related forms of learning and memory[J].Nat Protoc,2006,1(2):848-858.
[12]Deacon RM,Rawlins JN.T-maze alternation in the rodent[J].Nat Protoc,2006,1(1):7-12.
[13]Senior SL,Ninkina N,Deacon R,et al.Increased striatal dopamine release and hyperdopaminergic-like behaviour in mice lacking both alpha-synuclein and gamma-synuclein[J].Eur JNeurosci,2008,27(4):947-957.
[14]Ehehalt R,Keller P,Haass C,et al.Amyloidogenic processing of the Alzheimer beta-amyloid precursor protein depends on lipid rafts[J].J Cell Biol,2003,160(1):113-123.
[15]Dutheil F,Beaune P,Loriot MA.Xenobiotic metabolizing enzymes in the central nervous system:Contribution of cytochrome P450 enzymes in normal and pathological human brain[J].Biochimie,2008,90(3):426-436.
[16]Zhu L,Jie X,Liu AT,et al.Response of NG2 positive cells and other glial cells in the microenvironment of facial nucleus after facial nerve axotomy[J].Med J Chin PLA,2017,42(3):196-201.[朱鴷,结祥,刘安堂,等.面神经离断伤后核团微环境中NG2阳性细胞及各类胶质细胞的反应[J].解放军医学杂志,2017,42(3):196-201.]
[17]Holmes C.Review:systemic inflammation and Alzheimer's disease[J].Neuropath Appl Neuro,2013,39(1):51-68.
[18]Carter SF,Scholl M,Almkvist O,et al.Evidence for astrocytosis in prodromal Alzheimer disease provided by 11C-deuterium-Ldeprenyl:a multitracer PET paradigm combining 11C-Pittsburgh compound B and 18F-FDG[J].J Nucl Med,2012,53(1):37-46.
[19]Medeiros R,LaFerla FM.A strocytes:conductors of the Alzheimer'disease neuroinflammatory symphony[J].Exp Neurol,2013,239:133-138.
[20]Liu JJ,Cao N,Di JL,et al.Role of bone marrow mesenchymal stem ceils in repair of neural cell injury induced by okadaic acid[J].Med J Chin PLA,2017,42(5):377-382.[刘嘉婧,曹宁,翟晶磊,等.骨髓间充质干细胞对冈田酸致神经细胞损伤的修复作用[J].解放军医学杂志,2017,42(5):377-382.]
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