目的 本实验旨在探讨阿魏酸(FA)通过修复线粒体分裂-融合失衡改善阿尔茨海默病(AD)小鼠学习记忆障碍以及神经元保护的作用。方法 将KM小鼠随机分为空白对照组(A组)、模型对照组(B组)、阳性对照组(石杉碱甲片,C组)和低剂量阿魏酸模型组(D-低组)、高剂量阿魏酸模型组(D-高组),每组各10只。除A组小鼠外,其余小鼠一次性侧脑室注射Aβ1-42建立AD模型。采用自发活动实验和Morris水迷宫法检测FA对小鼠行为学的影响。采用PCR法检测线粒体动力学基因的mRNA水平。检测FA对AD病理标记蛋白以及线粒体分裂-融合蛋白表达的影响。另外,采用组织免疫荧光检测Aβ在大脑皮层神经元中的分布。结果 ①通过水迷宫实验证实,D-高组小鼠寻找站台的潜伏时间明显短于B组小鼠,但是仍然略微长于A组小鼠(P<0.05);但是D-高组小鼠在第4象限停留时间明显长于B组(P<0.05),与A组基本一致(P>0.05)。②D-高组小鼠大脑皮层组织中Drp1、钙调神经磷酸酶(calcineurin,CaN)催化亚单位α(CnAα)、CnAβ mRNA明显低于B组,但是仍然高于A组小鼠(P<0.05)。③D-高组小鼠大脑皮层组织蛋白中APP、Bace1、总Tau(Tau46)蛋白及S396 位点磷酸化的Tau(pS396)蛋白表达明显低于B组,但是仍然高于A组(P<0.05)。④除Drp1蛋白外,D-高组小鼠大脑皮层组织神经元细胞中Drp1Ser637、CnAα、蛋白激酶A的催化亚基c(PKAc)、线粒体融合蛋白基因 2(Mfn2)水平均与A组小鼠神经元趋于一致。⑤D-高组小鼠大脑皮层神经元中Aβ的形成和聚集较B组明显减少。结论 阿魏酸可以通过修复线粒体分裂-融合动力学失衡改善AD的病理损伤。
Abstract
OBJECTIVE To discuss the effect of ferulic acid(FA) on learning and memory impairment and neuron protection by repairing the imbalance of mitochondrial fission-fusion dynamics in Alzheimer′s disease (AD) mice. METHODS The KM mice were randomly divided into normal control group (A group, n=10), model group (B group, n=10), positive control group (huperzine A tablets, C group, n=10) and low dose of FA group (D-low group, n=10), high dose of FA group (D-high group, n=10). Mice in B, C, D-low and D-high groups were built as AD model by injecting Aβ1-42 into the lateral ventricle. The learning and memory ability of mice were detected by Morris water maze test. The mRNA of dynamin-related protein 1 (Drp1) were detected by PCR. The AD related pathological proteins and mitochondrial fission-fusion proteins were detected by Western blot. The content and distribution of Aβ was analyzed using immunofluorescence staining. RESULTS ①The escape latency of mice in D-high group was shorter than B group, but a little longer than A group (P<0.05). The swimming time in the 4th quadrant in D-high group was longer than B group (P<0.05), but as the same as A group (P>0.05). ②The mean expressions of Drp1, CaN subunit α (CnAα), CnAβ mRNA in D-high group were lower than B group, but higher than A group (P<0.05). ③The mean expressions of amyloid precursor protein (APP), beta-site APP cleaving enzyme 1 (Bace1), Tau46 and pS396 proteins in D-high group were lower than B group, but higher than A group (P<0.05).④The mean expressions of Drp1Ser637, CnAα, protein kinase A catalytic subunit c (PKAc), mitofusin gene 2(Mfn2) proteins in D-high group were basically identical with A group. ⑤The levels of Aβ formation and accumulation in mice cortex and hippocampus of D-high group were less than B group. CONCLUSION It′s suggested that ferulic acid(FA) might repair pathological damage of Alzheimer′s disease by improving the imbalance of mitochondrial fission-fusion dynamics.
关键词
阿魏酸 /
线粒体分裂-融合 /
阿尔茨海默病 /
动力学相关蛋白1 /
β-淀粉样蛋白
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Key words
ferulic acid /
mitochondrial fission-fusion imbalances /
Alzheimer′s disease /
dynamin-related protein 1 /
β-amyloid protein
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中图分类号：
R96
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参考文献
[1] SHEN Y, ZHANG Q, ZHAO S J, et al. Common pathological mechanism of Alzheimer′s disease and Parkinson′s disease. Chin Arch Tradit Chin Med(中华中医药学刊), 2018, 37(2):319-322.
[2] WANG Y P, ZHAI J B, ZHU F, et al. Prevalence of Alzheimer′s disease and its influencial factors among elderly people in communities. Chin J Public Health(中国公共卫生), 2011, 27(7):827-828.
[3] LONG Y X, TANG Y M, TANG L J, et al. Meta-analysis on major risk factors of Alzheimer′s disease in China. Chin Prev Med(中国预防医学杂志),2013, 14(1):59-63.
[4] JAKKI S L, SENTHIL V, YASAM V R, et al. The blood brain barrier and its role in Alzheimer′s therapy: an overview . Curr Drug Targets, 2018, 19(2):155-169.
[5] ZHANG G, WU J J, JIANG M, et al. Development of experiment on treating Alzheimer′s disease with traditional Chinese medicine. Chin J Exp Tradit Med Form(中国实验方剂学杂志), 2014,20(6):217-222.
[6] WANG Y, WANG X, YU S, et al. Effects of ferulic acid on oxidative stress and apoptosis related proteins in Alzheimer′s disease transgenic mice. Nat Prod Res Dev(天然产物研究与开发), 2017, 29(5):762-766.
[7] SCHMITZ T W, MUR M, AGHOURIAN M, et al. Longitudinal Alzheimer′s degeneration reflects the spatial topography of cholinergic basal forebrain projections. Cell Rep, 2018, 24(1):38-46.
[8] WU J L, SHEN M M, YANG S X, et al. Inhibition effect of ferulic acid on inflammation of microglia. Chin Pharmacol Bull(中国药理学通报), 2015, 31(1):97-102.
[9] LUO Y, ZHAO H P, ZHANG J, et al. Effect of ferulic acid on learning and memoryimpairments of vasculardementia rats and its mechanism of action. Acta Pharm Sin(药学学报), 2012, 47(2):256-260.
[10] MORI T, KOYAMA N, TAN J, et al. Combination therapy with octyl gallate and ferulic acid improves cognition and neurodegeneration in a transgenic mouse model of Alzheimer′s disease. J Biol Chem, 2017, 292(27):11310-11325.
[11] AKHTER H, HUANG W T, VAN GROEN T, et al. A small molecule inhibitor of plasminogen activator inhibitor-1 reduces brain amyloid-β load and improves memory in an animal model of Alzheimer′s disease. J Alzheimers Dis, 2018, 64(2):447-457.
[12] FURCILA D, DEFELIPE J, ALONSO-NANCLARES L. A study of amyloid-β and phosphotau in plaques and neurons in the hippocampus of Alzheimer′s disease patients. J Alzheimers Dis, 2018, 64(2):417-435.
[13] XU L L, SHEN Y, WANG X, et al. Mitochondrial dynamics changes with age in an APPsw/PS1dE9 mouse model of Alzheimer′s disease. Neuroreport, 2017, 28(4):222-228.
[14] JOSHI A U, SAW N L, SHAMLOO M, et al. Drp1/Fis1 interaction mediates mitochondrial dysfunction, bioenergetic failure and cognitive decline in Alzheimer′s disease. Oncotarget, 2017, 9(5):6128-6143.
[15] GUO M Y, SHANG L, HU Y Y, et al. The role of Cdk5-mediated Drp1 phosphorylation in Aβ1-42 induced mitochondrial fission and neuronal apoptosis. J Cell Biochem, 2018, 119(6):4815-4825.
[16] LEE S Y, CHO W H, LEE Y S, et al. Impact of chronic stress on the spatial learning and GR-PKAc-NF-κB signaling in the hippocampus and cortex in rats following cholinergic depletion. Mol Neurobiol, 2018, 55(5):3976-3989.
[17] KURUVA C S, MANCZAK M, YIN X, et al. Aqua-soluble DDQ reduces the levels of Drp1 and Aβ and inhibits abnormal interactions between Aβ and Drp1 and protects Alzheimer′s disease neurons from Aβ- and Drp1-induced mitochondrial and synaptic toxicities. Hum Mol Genet, 2017, 26(17):3375-3395.
[18] JIANG S, NANDY P, WANG W, et al. Mfn2 ablation causes an oxidative stress response and eventual neuronal death in the hippocampus and cortex. Mol Neurodegener, 2018, 13(1):5.
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脚注
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基金
国家自然科学基金项目资助(81671068)
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