模式动物斑马鱼在神经退行性疾病研究中的应用

斑马鱼作为一种新型模式低等脊椎动物,具有发育周期短、体外受精、胚胎透明、突变种多等优势,被广泛应用于神经、心血管、消化等方面的研究中。神经退行性疾病包括帕金森病、阿尔茨海默病等,近年来发病率不断上升,且有年轻化的趋势,严重影响人们的身体健康和生活质量。因其发病机制复杂,至今仍缺乏治疗此类疾病的方法。本文综述近年来用斑马鱼制作神经退行性疾病模型的主要方式及可行性,为研究神经退行性疾病中斑马鱼模型的选择提供参考。     

斑马鱼模型在中枢神经系统疾病中的研究进展

中枢神经系统（Central Nervous System,CNS）疾病严重损伤人的身心健康,在全球范围内造成巨大的疾病负担,且患病人数逐年呈增长趋势。由于CNS疾病的复杂性、多因素性及多基因调控性,导致相关疾病的发病机制尚不明确,市场上也严重缺乏针对该类疾病的有效药物。因此,探索CNS疾病的病理机制对药物的开发十分重要,急需一种模式生物来构建CNS疾病模型。斑马鱼（Danio rerio）作为一种重要的模式生物,不仅具有与人类高度保守的大脑组织区域,且具有丰富的社会学行为,成为研究CNS疾病和药物研发的重要工具。本文综述了近年来斑马鱼在CNS疾病研究中的相关研究进展,同时也介绍了一些小分子化合物在斑马鱼疾病模型中的研究应用,为今后CNS疾病的研究和相关药物的研发提供了基础。     

基于斑马鱼模型的鹿胶益气补血相关功效研究

目的 通过斑马鱼模型快速观察鹿胶补血、健骨、抗衰老等补益作用。方法 利用草甘膦(Gly)构建斑马鱼成鱼红细胞损伤模型,通过甲氨蝶呤(MTX)构建斑马鱼幼鱼造血功能损伤模型。通过泼尼松龙(Pred)构建斑马鱼幼鱼骨形成抑制模型。通过邻苯二甲酸二丁酯(DBP)构建斑马鱼幼鱼眼部细胞凋亡模型。结果 鹿胶可改善Gly诱导的斑马鱼成鱼红细胞核异常,促进造血因子SCL、GATA1的表达;改善Pred诱导的斑马鱼幼鱼骨形成抑制,促进成骨相关ALP、Runx2a的表达;减少DBP引起的斑马鱼幼鱼眼部细胞凋亡数,促进抗凋亡因子Bcl-2的表达。结论 鹿胶具有较好的保护红细胞、促进造血、促进骨形成、抗凋亡等作用,这些作用可能与其补血、抗骨质疏松、抗衰老等功效有关,该研究为鹿胶的临床应用提供科学依据,为进一步开发应用奠定基础。     

斑马鱼炎症模型及其在中药抗炎领域的应用

中药抗炎在近年来已成为一个重要的研究领域,中药制剂的抗炎临床研究已经取得了一定进展,但具体活性成分及作用机制仍需进一步明确。斑马鱼由于其独特的生物学优势,目前已被广泛应用于中药抗炎活性的筛选。综合近年来国内外有关斑马鱼模型在中药抗炎领域的报道,从已构建的斑马鱼炎症模型、斑马鱼模型中的炎症介质、斑马鱼模型在中药抗炎药物筛选中的应用3个方面对斑马鱼炎症模型及其在中药抗炎领域的研究进展进行综述,以期为模式生物斑马鱼在中药抗炎领域的研究应用提供新思路,为抗炎中药的开发与利用提供借鉴参考。     

抗肿瘤新药研究的新型模式生物斑马鱼

斑马鱼（Danio rerio），是一种用于研究脊椎动物胚胎学和发育遗传学的模式生物。近年来，利用斑马鱼来建立人类疾病研究模型已成为新的研究亮点。笔者就斑马鱼作为一种抗肿瘤新药研究模式生物所具有的优势、应用研究及其前景做一综述。     

基于模式生物斑马鱼模型的中药药效物质筛选技术及应用

目的作为一个复杂成分体系,中药药效物质的精准辨识和筛选是中药现代化研究的一个瓶颈问题,探索新的实验技术十分重要。方法斑马鱼是新兴的模式生物,具有体外受精、胚胎透明,不需要解剖可方便在显微镜下观察组织器官的发育等生物学优势,可在细胞板中完成实验,需要样品量少,已被欧洲替代法验证中心推荐为新的替代动物。斑马鱼全基因组测序结果显示斑马鱼与人的基因组相似性达87%,大量的研究表明斑马鱼对药物反应与临床反应也高度相似,为斑马鱼应用于研究人类疾病与药物筛选奠定了理论和实践基础。本实验室利用斑马鱼模型构建了较为齐全的中药药效物质筛选的技术体系,并对甘草、阿胶、栀子、乳香等传统中药进行药效或毒性成分的筛选辨识研究。结果本实验室利用斑马鱼模型建立了可涵盖肿瘤、心血管、神经系统、骨骼、皮肤、重要器官等的活性/毒性筛选技术平台。从前期大规模的筛选中,发现甘草具有一定的抗肿瘤作用,进一步利用斑马鱼活性跟踪技术和高速逆流色谱分离技术,发现了2个未报道的抑制血管生成的活性成分。新阿胶是山东福胶集团的独家品种,是以猪皮为来源,作为阿胶的替代品,但是现代药理学数据缺乏导致市场认可度低,因此我们利用斑马鱼模型系统开展了新阿胶的化疗损伤后的"扶正"与"生血"的作用机制。中药栀子临床发现具有防治血栓的功效,但是抗血栓的药效标志物并不清楚,作者采用斑马鱼模型整合代谢组学技术,发现3个代表性的抗血栓成分。乳香在历代典籍中具有"孕妇慎服"记载,但是原因不明,我们利用斑马鱼模型构建发育毒性模型,并结合灰色关联度分析法,证明乳香具有一定的发育毒性,并发现了潜在的毒性靶器官,并发现了主要的毒性效应成分。结论斑马鱼作为一种新型的模式生物,在中药药效物质筛选、活性评价、毒性预警方面都具有广阔的应用前景。     

新型模式生物斑马鱼及其胚胎在人类疾病模型研究中的应用

斑马鱼作为一种理想的模式动物在遗传学,发育生物学的研究中应用越来越广泛。由于斑马鱼具有容易获得,易于饲养,生殖周期短,繁殖能力强等特点,且其基因组测序的完成及与人类的高度保守性,使得斑马鱼及其胚胎在关于人类疾病的研究中有着重要的应用和实际价值。本文就斑马鱼及其胚胎在人类疾病研究中的应用作一简要概述。     

基于代谢组学和斑马鱼模型探究西洋参抗疲劳的关键活性成分

探索西洋参皂苷类成分中抗疲劳关键活性成分。利用斑马鱼模型评价西洋参样本的抗疲劳活性;运用代谢组学技术分析鉴定不同产地西洋参中主要皂苷类差异成分;通过构建西洋参皂苷类成分-疾病靶点PPI蛋白互作网络初步筛选出西洋参发挥抗疲劳作用的活性物质和相关靶点,利用分子对接的方法筛选出关键活性成分;最后使用斑马鱼模型评价关键活性成分的抗疲劳活性,动物实验获得山东省医学科学院伦理委员会的批准(SYXK20220005)。斑马鱼活性评价结果显示,两个产地的西洋参样本的活性存在显著差异,进一步经过代谢组学检测、模式判别共鉴定出其中10个差异皂苷类成分可能与抗疲劳活性有关。借助成分-疾病靶点的PPI筛选的抗疲劳的核心靶点,结合类药性参数及分子对接技术发现其中pseudoginsenoside F11与5个靶点均具有较好的结合活性。经斑马鱼模型验证发现拟人皂苷F11具有显著的抗疲劳活性。本研究运用代谢组学和斑马鱼模型筛选出了西洋参发挥抗疲劳的关键活性物质,为西洋参皂苷类抗疲劳的进一步研究提供参考。     

斑马鱼肿瘤模型研究进展

斑马鱼作为独具特点的新兴实验动物,已被广泛应用于肿瘤发生发展、抗肿瘤药物开发和筛选、最佳治疗方案确定等许多方面的研究。该文主要介绍斑马鱼肿瘤模型分类、特点及应用优势,旨在帮助人们了解和合理应用斑马鱼的肿瘤模型。     

模式生物斑马鱼在心功能评价中的应用

斑马鱼繁殖力强、体积小、易饲养、发育快速、胚胎透明,与人类基因组同源性高,心脏结构和功能与哺乳类动物高度相似。基于斑马鱼模型的药物实验具有用药量少、周期短、指标易观察等优势,近年来斑马鱼模型在药物心血管安全性评价和心脏保护活性筛选领域得到了广泛应用。结合课题组前期研究及国内外文献,综述斑马鱼心功能评价应用进展,以期为斑马鱼在心血管药物安全性和活性评价中的应用提供参考。     

Sex‐specific characterization and evaluation of the Alzheimer's disease genetic risk factor sorl1 in zebrafish during aging and in the adult brain following a 100 ppb embryonic lead exposure

Developmental lead (Pb) exposure is suggested in laboratory studies to be a trigger for neurodegenerative diseases such as Alzheimer's disease (AD). Sortilin‐related receptor, L (DLR class) A repeats‐containing (SORL1 ) is a recently identified AD genetic risk factor. SORL1 has limited characterization in vertebrate models in comparison to other AD genetic risk factors. To characterize SORL1 further, protein sequence homology between humans, mice and zebrafish was analyzed and showed conservation of functional repeats and domain orientation. Next, spatial expression of sorl1 in zebrafish larvae was completed and diffuse expression in neural tissue that was not restricted to the brain was observed. Influences of sex and age on quantitative expression of sorl1 in the brain of adult zebrafish were then assessed. Sex‐specific alteration of sorl1 expression transpired during the aging process in females. The zebrafish was then utilized to investigate the impacts of a 100 ppb embryonic Pb exposure on sorl1 expression and other known AD genetic risk factors. Sex‐specific quantitative gene expression analysis was completed with adult zebrafish brain to compare those developmentally exposed to Pb or a control treatment, but no significant difference in sorl1 expression or other AD genetic risk factors was observed. Overall, this study provided characterization of sorl1 with changes in brain expression during aging being female‐specific. This finding is in agreement with females being more prone to the onset of AD, but analysis of additional AD genetic risk factors is needed to facilitate our understanding of the impact of a 100 ppb embryonic Pb exposure. Copyright © 2016 John Wiley & Sons, Ltd.     

Curcumin and neurodegenerative diseases: a perspective

Introduction: Curcumin, a dietary polyphenol found in the curry spice turmeric, possesses potent antioxidant and anti-inflammatory properties and an ability to modulate multiple targets implicated in the pathogenesis of chronic illness. Curcumin has shown therapeutic potential for neurodegenerative diseases including Alzheimer's disease (AD) and Parkinson's disease (PD).

Areas covered: This article highlights the background and epidemiological evidence of curcumin's health benefits and its pharmacodynamic and pharmacokinetic profile. Curcumin's ability to counteract oxidative stress and inflammation and its capacity to modulate several molecular targets is reviewed. We highlight the neuroprotective properties of curcumin including pre-clinical evidence for its pharmacological effects in experimental models of AD and PD. The bioavailability and safety of curcumin, the development of semi-synthetic curcuminoids as well as novel formulations of curcumin are addressed.

Expert opinion: Curcumin possesses therapeutic potential in the amelioration of a host of neurodegenerative ailments as evidenced by its antioxidant, anti-inflammatory and anti-protein aggregation effects. However, issues such as limited bioavailability and a paucity of clinical studies examining its therapeutic effectiveness in illnesses such as AD and PD currently limit its therapeutic outreach. Considerable effort will be required to adapt curcumin as a neuroprotective agent to be used in the treatment of AD, PD and other neurodegenerative diseases.     

Use of Zebrafish Genetic Models to Study Etiology of the Amyloid-Beta and Neurofibrillary Tangle Pathways in Alzheimer's Disease

The prevalence of neurodegenerative diseases is increasing globally, with an imperative need to identify and expand the availability of pharmaceutical treatment strategies. Alzheimer''s disease is the most common neurodegenerative disease for which there is no cure and limited treatments. Rodent models are primarily used in Alzheimer''s disease research to investigate causes, pathology, molecular mechanisms, and pharmaceutical therapies. However, there is a lack of a comprehensive understanding of Alzheimer''s disease causes, pathogenesis, and optimal treatments due in part to some limitations of using rodents, including higher economic cost, which can influence sample size and ultimately statistical power. It is necessary to expand our animal model toolbox to provide alternative strategies in Alzheimer''s disease research. The zebrafish application in neurodegenerative disease research and neuropharmacology is greatly expanding due to several vital strengths spanning lower economic costs, the smaller size of the organism, a sequenced characterized genome, and well described anatomical structures. These characteristics are coupled to the conserved molecular function and disease pathways in humans. The existence of orthologs for genes associated with Alzheimer''s disease in zebrafish is also confirmed. While wild-type zebrafish appear to lack some of the neuropathological features of Alzheimer''s disease, the advent of genetic editing technologies has expanded the evaluation of the amyloid and neurofibrillary tangle hypotheses using the zebrafish and exploration of pharmaceutical molecular targets. An overview of how genetic editing technologies are being used on the zebrafish to create models to investigate the causes, pathology, molecular mechanisms, and pharmaceutical targets of Alzheimer''s disease is detailed.     

白藜芦醇治疗阿尔茨海默病的研究进展

神经退行性疾病是一种慢性进展性疾病,其特点是中枢神经系统神经元的逐渐丧失。由于血脑屏障的存在,经典的抗炎药物如类固醇激素和非甾体类抗炎药,对神经系统疾病的治疗作用有限。因此,开发新的抗炎药物,对于预防和治疗神经系统疾病具有重要的意义。白藜芦醇是一种有很强活性的天然多酚类物质,目前研究已显示其具有心血管保护、神经保护、免疫调节、肿瘤的化学预防作用。近年来还发现其具有抗神经炎症作用,可用于治疗神经精神性疾病,如帕金森病、阿尔茨海默病(AD)和亨廷顿症等。综述白藜芦醇对AD的保护作用及其机制研究进展,为进一步推进白藜芦醇用于防治AD的研究提供参考。     

7,8-Dihydroxyflavone Prevents Synaptic Loss and Memory Deficits in a Mouse Model of Alzheimer's Disease

Synaptic loss in the brain correlates well with disease severity in Alzheimer disease (AD). Deficits in brain-derived neurotrophic factor/tropomyosin-receptor-kinase B (TrkB) signaling contribute to the synaptic dysfunction of AD. We have recently identified 7,8-dihydroxyflavone (7,8-DHF) as a potent TrkB agonist that displays therapeutic efficacy toward various neurological diseases. Here we tested the effect of 7,8-DHF on synaptic function in an AD model both in vitro and in vivo. 7,8-DHF protected primary neurons from Aβ-induced toxicity and promoted dendrite branching and synaptogenesis. Chronic oral administration of 7,8-DHF activated TrkB signaling and prevented Aβ deposition in transgenic mice that coexpress five familial Alzheimer''s disease mutations (5XFAD mice). Moreover, 7,8-DHF inhibited the loss of hippocampal synapses, restored synapse number and synaptic plasticity, and prevented memory deficits. These results suggest that 7,8-DHF represents a novel oral bioactive therapeutic agent for treating AD.     

基于秀丽隐杆线虫模型的中药及其有效成分抗阿尔茨海默病的药理研究进展

近年来,运用秀丽隐杆线虫模型对治疗阿尔茨海默病（AD）药物进行的研究与日俱增,通过线虫的直观表型及其深入的作用机制来探讨中药抗AD活性,从而实现药物的初步筛选。在此基础上深入研究,发现中药复方六味地黄丸、提取部位银杏叶提取物及有效成分石杉碱甲等均取得较好的临床疗效,秀丽隐杆线虫模型对中药资源研发具有一定的优势。综述利用线虫模型筛选的中药复方、提取部位、有效成分及其抗AD的作用机制,为抗AD中药新药研发提供思路和参考。     

Editorial Central & Peripheral Nervous Systems: Amyloid β Peptide in Alzheimer's Disease Pathology: Towards a Rational Basis for Drug Discovery?

The role of the 39 - 43 amyloid β peptide known as Aβ in the pathophysiology of Alzheimer's Disease (AD) is a continuing area of active research, with often confusing and contradictory results. Considerable in vitro work suggests that this peptide, which is produced from amyloid precursor protein (APP) and can be overproduced in familial forms of AD, is neurotoxic. Inhibition of either the production of Aβ or its cellular actions represents a viable approach to the exploration of novel neurorestorative therapies for AD. Transgenic models of Aβ overproduction are a key to understanding the in vivo role of the peptide. The elusiveness of such models appears to have been addressed by recent reports of two mouse models in which overexpression of Aβ has resulted in some of the characteristic hallmarks of AD. However, the ability to select compounds for evaluation in such models remains highly dependent on preliminary in vitro testing, in which the effects of synthetic Aβ in CNS cell lines or primary cultures are examined. Such studies have been notoriously inconsistent in terms of batch-to-batch variability in peptide toxicity. While the solvent, pH and age of Aβ solutions can affect the physical form of the peptide (which, in turn, can be correlated with the neurotoxic potential of Aβ batches), there is still no way by which reliably neurotoxic lots of Aβ can be obtained. As a consequence, information involving the in vitro effects of Aβ is questionable and does not represent a good basis for a drug discovery effort. This is compounded by the fact that studies related to Aβ in solution in vitro often do not take into account the potential presence of clearance chaperones, like apolipoprotein E, and ‘pathological’ chaperones, like α1-antichymotrypsin.     

阿尔茨海默病的纳米疗法研究进展

阿尔茨海默病(alzheimer's disease,AD)是一种中枢神经退行性疾病,在社会人口老龄化日益加剧的今天,AD患病率不断上升,其严重程度足以干扰人类的日常生活,危害人类的健康.目前AD的发病机制尚不明确,没有有效的药物可以治愈AD.血脑屏障(Blood brain barrier,BBB)是血液循环与中枢神经系统之间的生物屏障,药物不能穿过BBB,使其在治疗中枢神经系统疾病时有一定的局限性.纳米释药系统可以非侵入性地将药物递送至大脑,通过靶向药物递送,可以降低药物的毒性,增加药物的生物利用度.本文简述了AD发病的几种假说,介绍了与AD相关的纳米药物(Nanomedicines,NMs)种类和研究进展,对纳米递药技术在AD治疗策略中的应用进行阐述和列举,为AD的NMs研究提供思路和参考.     

卡巴拉汀治疗阿尔茨海默病的脑局部一致性研究

目的 通过局部一致性（regional homogeneity,ReHo）分析方法探究卡巴拉汀治疗阿尔茨海默病（Alzheimer''s disease,AD）的脑功能影像机制。方法 9例轻中度AD患者给予卡巴拉汀治疗24周,治疗前后分别进行全面的神经心理测评和静息态功能磁共振扫描。并选取年龄、性别、受教育年限与之相匹配的健康对照组9例进行静息态功能磁共振扫描,比较卡巴拉汀治疗前后AD患者大脑ReHo值的变化。结果 与治疗前比,AD患者治疗后,表现出简易智力状态检查量表评分增加（P<0.05）,老年痴呆症评估量表-认知部分和日常生活活动能力评分降低（P<0.05）;AD患者在左内侧额上回,左侧眶内额上回和右侧杏仁核表现出ReHo值减少（P<0.05）;此外,老年痴呆症评估量表-认知部分得分变化与左侧眶内额上回ReHo值呈正相关。结论 卡巴拉汀能够改善AD的认知功能,可能是通过与认知相关的内侧额上回和杏仁核的自发脑活动来实现的,相关脑区未来有望作为此类药物疗效监测的生物学标志物。