转基因线虫在阿尔茨海默症研究中的应用

李　梵,罗剑鸣,刘少莉,韩　蕾,卢　江,张雅丽

(中国农业大学食品科学与营养工程学院,北京 100083)



转基因线虫在阿尔茨海默症研究中的应用

李梵,罗剑鸣,刘少莉,韩蕾,卢江,张雅丽*

(中国农业大学食品科学与营养工程学院,北京 100083)

由于秀丽线虫基因与人类基因的同源性高达40%,其神经元的功能和神经递质与人类的也高度类似,因而秀丽线虫成为研究神经性疾病机制的有效模型。本文就转基因秀丽线虫在阿尔茨海默病中的研究进展进行了阐述,总结了近年来天然提取物在阿尔茨海默症中的应用,并对以转基因线虫为模型进行研究的单体进行了阐述。

秀丽线虫,阿尔茨海默症,天然提取物,单体

阿尔茨海默病(Alzheimer’sdisease,AD)是世界上引起死亡最多的十种疾病之一,世界上四千万人患病,在未来的十年这个数字还会增加[1]。大量的临床实验都在研究其病理学和病因学,除了一些缓解症状的药在使用之外,AD的病因及治愈并没有被完全阐述。研究人员设计了各种动物模型诸如β-淀粉样蛋白模型等[2]、胆碱能系统损伤模型[3]、铝模型[4]、tau蛋白模型[5]试图寻找发病原因及机制,并取得了一定的研究成果。秀丽线虫具有易培养、生长周期短、通体透明、体细胞数目一定、特定细胞位置恒定等优点。此外,秀丽线虫基因组测序也在1998年完成。随后,秀丽线虫作为模式生物在细胞凋亡、神经性疾病等方面研究中应用更加广泛。其中,在秀丽线虫中建立的AD模型主要有Aβ模型和tau模型两种。本文主要介绍Aβ模型线虫在AD中的研究应用。

1　阿尔茨海默病

AD是以进行性记忆减退、认知障碍与人格改变为主要临床表现的一种神经系统变性疾病,也是老年人群中最主要的神经退行性疾病[6]。AD包括一系列神经损伤的症状,氧化损伤,炎症和神经元机能障碍[7],并可见老年斑(Senile plaques,SP)与神经原纤维结(Neurofibrillary tangles,NFT)等病变[8]。AD的早期病理是大脑中β样淀粉蛋白前体代谢成为Aβ蛋白[9],Aβ的积累导致老年斑的形成。老年斑又称淀粉样斑(Amyloid Plaques,AP),主要由细胞外纤维和β淀粉样蛋白(beta-amyloid,Aβ)形成沉淀引起。目前Aβ是比较明确的致病因子。Aβ刺激下小胶质细胞的活化、还原型烟酰胺腺嘌呤二核苷酸(NADPH)氧化酶的激活、大量活性氧的生成、有害自由基的产生、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及诱导型一氧化氮合酶(induced nitric oxide synthase,iNOS)等各种促炎因子的释放,都直接或间接导致了神经系统的损伤[10]。大量体外实验研究表明,Aβ聚集形成纤维或低聚物均比单体结构具有更强的神经毒性[11]。因此,寻找某种阻止Aβ聚集的抑制剂被认为是一种延缓AD的治疗方案。

2　AD的秀丽线虫模型

在研究线虫的过程中,研究人员发现线虫中含有人类APP类似基因apl-1,它在线虫中是跨膜蛋白,但是它不含Aβ序列,因此需要通过转基因的方式来得到AD模型。1995年,Link CD通过转基因的方式建立了线虫的Aβ模型[12]。转基因线虫模型用来研究AD毒性机理[13]、Aβ引起的基因表达的变化[14-17]及探究药物抗AD的功效。

在众多实验室研究中,常用的转基因线虫品系主要为CL2006、CL4176等(表2)。其中CL4176使用肌肉细胞特异性启动子myo-3来控制Aβ的表达,使线虫表型上出现抽搐症状,复制了AD的症状和病理变化,从而可以明显地区分转基因的线虫。CL4176线虫不会在体内形成Aβ斑,因为Aβ在细胞内主要以水溶形式存在[18-19]。而CL2006线虫使用启动子unc-54来控制Aβ的表达,并引入表型基因rol-6。CL2006线虫同CL4176线虫一样会出现肌肉细胞中Aβ沉积,此外还会有进行性麻痹的表现。

3　线虫中AD的相关基因

3.1Aβ基因

虽然在转基因线虫中Aβ基因通过温度诱导表达或是结构表达使线虫体内Aβ大量形成,但Aβ基因的表达通常与β淀粉沉积并无直接联系[20-22]。这可能是由于不同实验所用转基因线虫品系不同,还与收集样品时间相关。

3.2乙酰胆碱酯酶基因

胆碱能缺失学说认为AD发病是源于病人体内乙酰胆碱的缺失[23]。由于线虫基因的保守性,可以通过检测线虫乙酰胆碱酯酶的表达量观察给药的效果。与脊椎动物只有一个乙酰胆碱酯酶基因不同,线虫有4种乙酰胆碱酶基因,ace-1在所有的体细胞和外阴肌肉细胞中表达[24],ace-2仅在神经细胞中表达[25],ace-3则是在眼部肌肉细胞和神经细胞中表达,ace-4基本检测不到[26]。但ACE-1、ACE-2占了酶活的95%,ACE-3占了5%,ACE-4基本检测不到酶活[27]。

3.3抗炎与应激相关基因

与神经性疾病相关的促进炎症的基因及细胞素主要有TNFα,IL-1和IL-6。这些因子在不同的AD模型中已经证实与Aβ的神经毒性及由Aβ引起的AD发病机制有关[28-30]。Link CD等研究表明,在AD病人脑组织中TNFA1P1(TNFα-induced protein 1)表达上调。线虫中有TNFA1P1的同源基因F22E5.6和ZC239.12。线虫升温培养后,CL4176体内F22E5.6和ZC239.12表达上调[30]。

热激蛋白(heat shock proteins,HSPs)是一类类似分子伴侣的小分子多肽,在Aβ应激条件下促进其表达,增强抵抗力[31]。由于线虫中热激反应是由神经元主导的[32],HSP-16被认为是毒蛋白异常积累下的保护蛋白。在线虫体内的HSP-16.2过表达会抑制β淀粉样蛋白的毒性[33]。在AD病人脑组织中与应激相关的伴侣蛋白αB-crystallin(CRYAB)表达上调,而线虫CL4176中CRYAB同源基因HSP16-2(γ46H3A.D)和HSP16-4(γ46H3A.E)在升温培养后表达上调。六味地黄汤粗提物促进了热激蛋白HSP-16.2的表达,使得线虫瘫痪发生时间延后[34]。有研究表明姜黄素能够抑制IL-1、TNFα、COX-2、iNOS和NF-κB等炎症因子的表达[35]。

3.4衰老相关基因

AD是与衰老相关的神经性疾病[36]。衰老相关基因主要有早衰老基因ps-1和ps-2,胰岛素样生长因子-1基因等基因。在线虫中,与ps-1和ps-2同源的基因是sel-12[37]和hop-1[38]。

4　AD的治疗

由于AD发病机理的复杂性,引起AD症状的上游神经递质的缺失已经揭示[39],因此,越来越多的研究集中在开发新药品上,以调节上游靶点,并由此揭示AD的发病机制。

4.1药物研究

目前的治疗及病情控制主要依赖于早发现,尽早服用一些有助于缓解与病情相关的药物延迟病情的进一步恶化。

过去的十年中,许多药物对AD有一定的缓解作用,特别是对Aβ的形成或是Aβ的沉积[40]。乙酰胆碱酯酶抑制剂是目前市场上主要的治疗 AD 的药物。美国 FDA批准的五种抗 AD 药物中乙酰胆碱酯酶抑制剂占了四种,它们是tacrine,donepezil,rivastigmine,galanthamine,这些药物能够改善患者的症状,提高患者的认知能力,但是并不能完全的治愈 AD,只能暂时改善症状,一定程度延缓病程,远期疗效不好,而且多有较为严重的毒副作用[41]。有些药物会引起肠胃功能损害[28,42]。Memantine为一种美国食品药品管理局批准用于治疗中度和重度AD的药物,可以通过削弱N-甲基-D-天冬氨酸感受器调节胆碱能代谢平衡,但也对肠胃有副作用[43]。

目前临床上并没有一种单靶点的药物有效并安全。近年来,许多研究学者开始寻求多靶点药物,同时对AD致病因子中几个重要的作用靶点进行控制,这是一种高效的阻止AD神经退化进程的方法[44-48]。对此,有关体内研究实验表明,memoquin[43]和IQM-622[49]作为多靶点是有效的。Donepezil和hupine Y[50]的合成物,对官能团进行修饰,与乙酰胆碱酶相互作用,起到类胆碱功能。另一方面,多靶点药物结合乙酰胆碱酯酶使得乙酰胆碱酯酶无法再与Aβ结合[51-52],从而阻止了Aβ的聚合[53]。体外研究结果表明,合成物AVCRI104P4也能够抑制丁酰胆碱酯酶,从而抑制Aβ的聚集[39]。多靶点研究药物的设计考虑因素主要为多靶点药物对多个靶点作用强度的问题,力求对各个靶标作用均衡[54]。目前,多靶点药物仍在研究阶段,诸多研究者期待通过化学合成的方法制备有效的多靶点药物。

4.2植物提取物研究

近几年来,国内外抗AD的研究热点逐渐转移到天然提取物上,一是由于AD病理复杂,并没有某种单一药物具有良好效果;二是因为其中一些药物具有副作用。而在日常饮食中会摄入一些抗氧化剂,这些抗氧化剂对AD以及衰老相关的疾病具有一定的影响及作用。表1显示的为几种植物提取物的研究成果。

从表1中可以看出,很多为中药成分,大多是从中医的角度出发筛选药物并临床实践后发现的抗AD候选药物。其中,银杏提取物EGb761开启了植物提取物研究的热潮。银杏提取物对中枢神经系统的作用机理可能与氧化、清除脂质自由基,保护膜蛋白等作用有关[71]。用六味地黄汤的乙醇提取物(LWDH-EE)干预转基因秀丽线虫CL4176,发现其能有效延缓Aβ1-42诱导的秀丽线虫麻痹症状[34]。在胡桃酮诱导CL4176 氧化应激后,LWDH-EE能降低其体内活性氧的产生,在热应激后,LWDH-EE能减少CL4176中amy-1基因表达,同时升高HSP的表达。这说明LWDH-EE通过抗氧化、调节热休克蛋白的途径来缓解β淀粉样蛋白产生的毒性作用[34]。

表1　植物提取物抗AD及衰老研究

4.3单体研究

由于植物粗提物成分的复杂性,很难解释清楚抗AD的作用机制和有效成分。同时由于单体提取制备的困难性,目前只有一部分单体被用来进行抗AD发病机制的高水平研究。

单体物质也可以从结构上的相似性推断功能的相似性,丹酚酸与姜黄素结构上相似,姜黄素具有抗Aβ功能,通过实验证明丹酚酸也有抗Aβ聚集功能[72]。分子动力学模拟表明,丹酚酸与Aβ的C端结合从而稳定了Aβ的α螺旋结构。小分子可以与Aβ的C端结合使得构象由α螺旋变为β折叠[73-74]。体外实验研究显示,单体的取代基和空间构象共同影响抗Aβ活性[75]。但体外实验与体内实验并不完全一致,LWDH体外实验没有抑制Aβ聚集,在体内实验中却是有效的[34]。不同的单体作用机理不同。虽然部分生物碱也有抗氧化性质,但在线虫实验中,抗氧化性质在抗AD的过程并不是完全必需的。Rong Di等研究结果表明Haemanthidine等四种生物碱具有抑制Aβ诱导毒性的作用,而且通过ROS验证后发现,这4种生物碱对缓解Aβ诱导毒性的原因与抗氧化作用无关,可能是其它直接作用于Aβ及其相关基因上的途径来起到缓解Aβ诱导毒性的作用[27]。通过线虫瘫痪实验可以看出,这4种生物碱与治疗老年痴呆症药物memantine相比,四种生物碱的抑制作用更强。然而,Kumar等指出水飞蓟素抗AD与抗氧化有关[76]。表2总结了以秀丽线虫为研究工具能够对AD有缓解作用的单体研究。

表2　目前所研究抗AD的单体

虽然单体研究有助于确定某一物质的作用机理,但植物粗提物中多种物质的存在会相互影响,共同作用于AD致病因子,从而降低致病因子对机体的损害。绿茶提取物中水溶性VC与脂溶性VE成分之间会相互促进,更好地发挥抗AD的功能,而VE单独作用时效果不明显[68]。

5　展望

模式动物为解决天然产物对神经性疾病的治疗提供了便利。转基因线虫作为模式动物,用来探究Aβ的毒性的分子机理,在植物提取物抗氧化物乃至药物的筛选方面发挥着重要的作用,线虫已经成为研究衰老及与衰老相关的神经性疾病的良好模型。利用线虫已阐明了植物提取物在体内的抗AD部分机制,能够很好地促进该研究领域向高等动物继续深化,直至发展成为临床上治疗氧化应激相关疾病首选的纯天然药物。

虽然线虫模型已经取得了一定成果,但是线虫作为一种低级动物只能作为天然药物的初期筛选,之后仍需要在老鼠或猴子等高等动物中实验。其次,AD病因复杂,发病机制并未研究透彻,目前发现的与AD有关的基因位点很有限,发现新位点,明确更多与AD相关的基因才能更好地研发治疗药物。

[1]Prince M,Albanese E,Guerchet M,et al.World Alzheimer Report 2014:Dementia andRisk Reduction;Alzheimer’s Disease International:London,UK,2014.Available online:http://www.alz.co.uk(accessed on 20 February 2015.

[2]Prakash A,MedhiB,Chopra K.Granulocyte colony stimulating factor(GCSF)improves memory and neurobehavior in an amyloid-βinduced experimental model of Alzheimer’s disease[J].PharmacolBiochemBehav,2013,110:46-57.

[3]Kuznetsova E,Schliebs R.β-Amyloid,cholinergic transmission,and cerebrovascular system-adevelopmentalstudy in a mouse model of Alzheimer’s disease[J].Curr Pharm Des,2013,19(36):6749-6765.

[4]Nivsarkar M,Banerjee A.Establishing the probable mechanism of L-DOPA in Alzheimer’sdisease management[J].Acta Pol Pharm,2009,66(5):483-486.

[5]Lagoja I,Pannecouque C,Griffioen G,et al.Substituted 2-aminothiazoles are exceptional inhibitors of neuronal degeneration in tau-driven models of Alzheimer’s disease.[J].Eur J Pharm Sci,2011,43(5):386-392.

[6]Hardy J,Selkoe D J.The amyloid hypothesis of Alzheimer’s disease:progress and problems on the road to therpeuties[J].Science,2002,297(5580):353-356.

[7]Mähler A,Mandel S,Lorenz M,et al.Epigallocatechin-3-gallate:a useful,effectiveand safe clinical approach for targeted prevention and individualizedtreatment of neurological diseases[J].EPMA,2013,4(1):5-21.

[8]Brookmeyer R,Johnson E,Ziegler-Graham K,et al.Forcasting the global burden of Alzheimer’s disease[J].Alzheimer’s Dement,2007,3(3):186-191.

[9]Hardy J,Chartier-Harlin MC,Mullan M.Alzheimer’s disease:the newagenda[J].Am.J.Human Genet,1992,50(3):648-651.

[10]AskarovaS,Yang X,Sheng W,et al.Role of Abeta-receptor foradvanced glycation endproducts interaction in oxidative stress andcytosolic phospholipase activation in astrocytes and cerebralendothelial cells[J].Neuroscience,2011,199:375-385.

[11]Dahlgren KN,Manelli AM,Stine Jr,et al.Oligomeric and fibrillar species of amyloid-beta peptides differentially affectneuronal viability[J].Biol.Chem,2002,277(35):32046-32053.

[12]Link CD.Expression of human beta-amyloid peptide in transgenic Caenorhabditis elegans[J].Proc Natl AcadSci USA,1995,92(20):9368-9372.

[13]Link CD.Caenorhabditis elegans models of age-associated neurodegenerative diseases:lessons from transgenic worm models of Alzheimer’s disease[J].ExpGerontol,2006,41(10):1007-1013.

[14]Hassan WM,Dostal V,Huemann BN,et al.Identifying Aβ-specificpathogenic mechanisms using a nematode model of Alzheimer’s disease[J].Neurobiol.Aging,2015,36(2):857-866.

[15]Li J,Le W,Sashidhara KV,et al.Benzofuran-chalcone hybrids as potentialmultifunctional agents against Alzheimer’s disease:Synthesis and in vivo studies with transgenicCaenorhabditiselegans[J].ChemMedChem,2014,9(12):2671-2684.

[16]Cao Y,Wang L,Lin Z,et al.Dehydroabietylamine derivatives asmultifunctional agents for the treatment of Alzheimer’s disease[J].Med.Chem.Commun,2014,5:1736-1743.

[17]Cao YY,Wang L,Ge H,et al.Salvianolic acid A,a polyphenolicderivative from Salvia miltiorrhizabunge,as a multifunctional agent for the treatment ofAlzheimer’s disease[J].Mol.Divers,2013,17(3):515-524.

[18]DostalV,Link CD.Assaying beta-amyloid toxicity using a transgenic C.elegansmodel[J].J Vis Exp,2010,9(44):2252-2255.

[19]Drake J,Link CD,Butterfield DA.Oxidative stress precedes fibrillarydeposition of Alzheimer’s disease amyloid beta-peptide(1-42)in a transgenicCaenorhabditiselegansmodel[J].Neurobiol Aging,2003,24(3):415-420.

[20]Strayer A,Wu Z,Christen Y,et al.Expression of the smallheat-shock protein Hsp16-2 in Caenorhabditis elegans is suppressed by Ginkgobiloba extract EGb 761[J].Faseb J,2003,17(15):2305-2307.

[21]Dostal V,Roberts CM,Link CD.Genetic mechanisms of coffee extractprotection in a Caenorhabditis elegansmodel of beta-amyloid peptide toxicity[J].Genetics,2010,186(3):857-866.

[22]Arya U,Dwivedi H,Subramaniam JR.Reserpine ameliorates Abetatoxicity in the Alzheimer’s disease model in Caenorhabditis elegans[J].ExpGerontol,2009,44(6-7):462-466.

[23]Wenhai H,Li T,Ying S.Searching for the multi-target-directed ligands against Alzheimer’s disease:discovery of quinoxaline-based hybrid compounds with AChE,H3R and BACE 1 inhibitory activities[J].Bioorgan Med Chem,2011,19(23):7158-7167.

[24]Culetto E,Combes D,Fedon Y,et al.Structureandpromoter activity of the 59 flanking region of ace-1,the gene encodingacetylcholinesterase of class A in Caenorhabditis elegans[J].Mol Biol,1999,290(5):951-966.

[25]Combes D,Fedon Y,Toutant JP,et al.Multiple acegenesencodingacetylcholinesterases of Caenorhabditis elegans have distinct tissueexpression[J].Eur J Neurosci,2003,18(3):497-512.

[26]Selkirk ME,Lazari O,Hussein AS,et al.Nematodeacetylcholinesterases are encoded by multiple genesand perform nonoverlappingfunctions[J].ChemBiol Interact,2005,8(157-158):263-268.

[27]RongDi,Lijuan Xin,RitupriyaYamujala,et al.Acetylcholineestarase-inhibiting alkaloids fromLycorisradiatedelay paralysis of amyloid beta-expressing transgenic C.elegans CL4176[J].PLoS One,2013,8(5):e63874.

[28]Tabet N.Acetylcholinesterase inhibitors for Alzheimer’s disease:anti-inflammatoriesin acetylcholine clothing[J].Age Ageing,2006,35(4):336-338.

[29]Reale M,Iarlori C,Gambi F,et al.Acetylcholinesterase inhibitors effects on oncostatin-M,interleukin-1 beta andinterleukin-6 release from lymphocytes of Alzheimer’s disease patients[J].ExpGerontol,2005,40(3):165-171.

[30]Link CD,Taft A,Kapulkin V,et al.Gene expressionnanalysis in a transgenic Caenorhabditis elegansAlzheimer’s disease model[J].Neurobiol Aging,2003,24(3):397-413.

[31]Link CD,Cypser JR,Johnson CJ,et al.Direct observation ofstress response in Caenorhabditis elegans using a reporter transgene[J].Cell StressChaperones,1999,4(4):235-242.

[32]Prahlad V,Cornelius T,Morimoto RI.Regulation of the cellular heatshock response in Caenorhabditis elegans by thermosensoryneurons[J].Science,2008,320(5877):811-814.

[33]Fonte V,Forrestal M,Wagner E,et al.Suppressionof in vivo beta-amyloid peptide toxicity by overexpression of the HSP-16.2 smallchaperoneprotein[J].BiolChem,2008,283(2):784-791.

[34]Sangha JS,Sun X,Wally OS,et al.LiuweiDihuang(LWDH),atraditional Chinese medicinal formula,protects againstβ-amyloidtoxicity in transgenic Caenorhabditis elegans[J].PLoS One,2012,7(8):e4399.

[35]ColeGM,Lim GP,Yang F,et al.PreventionofAlzheimer’sdisease:Omega-3 fattyacidandphenolicanti-oxidantinterventions[J].NeurobiolAging,2005,26(1):133-136.

[36]Miksys SL,Tyndale RF.Neurodegenerative diseases:a growing challenge[J].ClinPharmacolTher,2010,7(8):427-430.

[37]LevitanD,Greenwald I.Facilitation of lin-12-media ted signalling by sel-12,a CaenorhabditiselegansS182 Alzheimer’sdisease gene[J].Nature,1995,377(6547):351-354.

[38]LiX,Greenwald I.HOP-l,a Caenorhabditiseleganspresenilin,appears to be functionally redundant with SEL-12 presenilinand to facilitate LIN-12 and GLP-1 signaling[J].Proc NatiAcadSci USA,1997,94(22):12204-12209.

[39]Irene Sola,ElisabetViayna,Tània Gómez,et al.Synthesis and in vivo efficacy studies of a novelmultitargetanti-Alzheimer’s compound[J].Molecules,2015,20(3):4492-4515

[40]Citron M.Strategies for disease modification in Alzheimer’s disease[J].Nat Rev Drug Discov,2004,5(9):677-685.

[41]Youdim M B,Buccafusco J J.CNS targets for multi-functional drugs in the treatment of Alzheimer’s and Parkinson’s disease[J].Neural Transm,2005,112(4):519-537.

[42]McGleenon BM,Dynan KB,Passmore AP.Acetylcholinesteraseinhibitors in Alzheimer’s disease.[J].Br J ClinnPharmacol,1999,48(4):471-480.

[43]Wenk GL,Quack G,Moebius HJ,et al.No interaction ofmemantine with acetylcholinesterase inhibitors approved for clinical use[J].Life Sci,2000,66(12):1079-1083.

[44]Lopes JP,Bertorelli R,Tarozzo G,et al.Pharmacological characterization of memoquin,a multitargetcompoundfor the treatment of Alzheimer’s disease[J].PLoS One,2013,8(2):e56870.

[45]Guzior N,Więckowska A,Panek D,et al.Recent development of multifunctionalagents as potential drug candidates for the treatment of Alzheimer’s disease[J].Curr Med Chem,2015,22(3):373-404.

[46]GeldenhuysWJ,Van der,SchyfCJ.Rationally designed multi-targeted agents againstneurodegenerativediseases[J].Curr Med Chem,2013,20(13):1662-1672.

[47]Chen X,Decker M.Multi-target compounds acting in the central nervous system designed fromnatural products[J].Curr Med Chem,2013,20(13):1673-1685.

[48]Russo P,Frustaci A,Del Bufalo A,et al.Multitarget drugs of plants originacting on Alzheimer’s disease[J].Curr Med Chem,2013,20(13):1686-1693.

[49]Bolos M,Spuch C,Pascual C,et al.Effects of a tacrine-8-hydroxyquinoline hybrid(IQM-622)onAβaccumulation and cell death:Involvement in hippocampal neuronal loss in Alzheimer’sdisease[J].Neurobiol Dis,2012,46(3):682-691.

[50]Viayna E,Galdeano C,Ratia M,et al.Novel huprine derivatives with inhibitory activity towardβ-amyloidaggregation and formation as disease-modifying anti-Alzheimer drug candidates[J].ChemMedChem,2010,5(11):1855-1870.

[51]Inestrosa NC,Alvarez A,Moreno RD,et al.Acetylcholinesterase accelerates assembly of amyloid-β-peptides intoAlzheimer’s fibrils:Possible role of the peripheral site of the enzyme[J].Neuron,1996,16(4):881-891.

[52]Canales MA,Shin I,Weiner LM,et al.A structuralmotif of acetylcholinesterase that promotes amyloid beta-peptide fibril formation[J].Biochemistry,2001,40(35):10447-10457.

[53]Castro A,Martinez A.Targeting beta-amyloid pathogenesis through acetylcholinesteraseinhibitors[J].CurrPharm Des,2006,12(33):4377-4387.

[54]夏亮.多靶点AD药物的设计与合成[D].南宁:广西医科大学,2014.

[55]Gutierrez-Zepeda A,Santell R,Wu Z,et al.Soy isoflavoneglycitein protects against beta amyloid-inducedtoxicity and oxidative stress in transgenic Caenorhabditis elegans[J].BMC Neuroscience,2005,6:54.

[56]Wu Y,Wu Z,Butko P,et al.Amyloid-betainducedpathological behaviors are suppressed by Ginkgo bilobaextract EGb761 and ginkgolides in transgenic Caenorhabditis elegans[J].Neurosci,2006,26(50):13102-13113.

[57]LiangYQ,TangXC.ComparativeeffectsofhuperzineA,donepezil and rivastigmine on corticalacetylcholinelevelandacety lcholinesteraseactivity in rats[J].Neuro SciLett,2004,361(1-3):56-59.

[58]张云玲,刘东梅,吴庆四，等.黄芪提取物对Aβ25-35所致阿尔茨海默病模型大鼠的学习记忆能力及海马神经元Bcl-2和Bcl-xl表达的影响[J].安徽医科大学学报,2007,42(3):299-303.

[59]叶翠飞,楚晋,李林，等.淫羊藿黄酮对APP转基因小鼠学习记忆及β-amyloid生成的影响[J].中国科学技术大学学报,2008,38(4):439-448.

[60]陈烈,黄君英,薛俐.复方何首乌浸膏治疗阿尔茨海默病的疗效研究[J].中南大学学报,2010,35(6):612-616.

[61]Soodi M,Hajimehdipoor H,Soleimani M,et al.Comparison of neuroprotective effects of melissa officinalis total extract and its acidic and non-acidic fractions against Aβ-induced toxicity[J].Iran J Pharm Res,2013,12(2):415-423.

[62]Su T,Xie S,Wei H,et al.Synthesis and biological evaluationofberberine-thiophenyl hybrids as multi-functional agents:inhibition ofacetylcholinesterase,butyrylcholinesterase,and Abetaaggregation and antioxidantactivity[J].Bioorg Med Chem,2013,21(18):5830-5840.

[63]Xian YF,Lin ZX,Mao QQ,et al.Bioassayguided isolation of neuroprotectivecompounds from uncariarhynchophyllaagainst beta-amyloid-inducedneurotoxicity[J].Evid Based Complement AlternMed,2012:802625.

[64]崔理立.针对阿尔茨海默病中Aβ42靶点的天然药物及其分子机制的研究[D].长春:吉林大学,2012.

[65]董晓华.蛇床子素对AD大鼠学习记忆的影响及机制研究[D].石家庄:河北医科大学,2012.

[66]Chul-Kyu Kim,Jin-Kook Park,et al.Increasedresistance to stress and an anti-aging effect due toAcanthopanaxsessiliflorusroots in Caenorhabditis elegans[J].Food SciBiotechnol,2014,23(5):1653-1659.

[67]张振清.红景天苷对Aβ1-40所致AD实验模型的干预作用及机制研究[D].石家庄:河北医科大学,2014.

[68]Atsushi Takahashi,Tatsuro Watanabe,Takashi Fujita,et al.Green tea aroma fraction reducesβ-amyloid peptide-induced toxicity in Caenorhabditis eleganstransfected with humanβ-amyloid minigene[J].Bioscience Biotechnology and Biochemistry,2014,78(7):1206-1211

[69]熊哲,袁沛,童杰文，等.红茶预防及调控阿尔茨海默病作用研究进展[J].食品安全质量检测学报,2015,6(4):1219-1224.

[70]Jiang X,Jia LW,Li XH,et al.Capsaicin ameliorates stress-induced Alzheimer’sdisease-like pathological and cognitive impairments in rats[J].Alzheimers Dis,2013,35(1):1206-1211.

[71]Boveris AD,Galleano M,Puntarulo S.In vivo supplementation with Ginkgo biloba protects membranes against lipid peroxidation[J].Phytother Res,2007,21(8):735-740.

[72]ColesM,BicknellW,Watson AA,et al.Solutionstructure of amyloid beta-peptide(1-40)in a water-micelleenvironment:Is the membrane-spanning domain where we think itis?[J].Biochemistry,1998,37(31):11064-11077.

[73]Yang C,Zhu XL,Li JY,et al.Exploration of the mechanismfor LPFFD inhibiting the formation of beta-sheet conformationof A beta(1-42)in water[J].Mol Model,2010,16(4):813-821.

[74]Wang B,Liu JX,Meng HX,et al.Blocking effectofsalvianolic acid A on calcium channels in isolated rat ventricularmyocytes[J].Chin J Integr Med,2012,18(5):366-370.

[75]Riviere C,Papastamoulis Y,Fortin PY,et al.New stilbene dimers against amyloid fibril formation[J].Bioorganic MedChemLett,2010,20(11):3441-3443.

[76]Sonani RR,Singh NK,Prasad B,et al.Phycoerythrin extends life span and health spanofCaenorhabditis elegans[J].AGE,2014,36(5):9717.

[77]张健炜,熊哲,高燕,等.红茶延长秀丽线虫在环境氧化应激中寿命的研究[J].茶业科学,2013,33(4):295-300.

[78]何露,李中,韦睿,等.姜黄素类化合提高线虫的抗氧化应激能力[J].中国病理生理杂志,2014,30(1):154-158.

[79]McColl G,Roberts BR,Gunn AP,et al.TheCaenorhabditiselegansA beta 1-42 model of Alzheimer disease predominantlyexpresses A beta 3-42[J].BiolChem,2009,284(34):22697-22702.

[80]Luisa Diomede,Giuseppe Cassata,Fabio Fiordaliso,et al.Tetracycline and its analogues protectCaenorhabditis elegansfromβamyloid-induced toxicity by targeting oligomers[J].Neurobiology of Disease,2010,40(2):424-431.

[81]Diomede L,Rigacci S,Romeo M,et al.Oleuropeinagly coneprotects transgenic C.elegansstrainsexpressing Abeta-42 by reducing plaque load and motor deficit[J].PLoS ONE,2013,8(3):e58893.

[82]Martorell P,Bataller E,Llopis S,et al.A cocoa peptide Protects Caenorhabditis elegans from oxidative stress and beta-amyloid peptide toxicity[J].PLoS ONE,2013,8(5):e63283.

[83]Alavez S,Vantipalli MC,Zucker DJ,et al.Amyloid-binding compounds maintain protein homeostasis duringageing and extend lifespan[J].Nature,2011,472(7342):226-229.

[84]Kumar J,Park KC,Awasthi A,et al.Silymarin extends lifespan and reduces proteotoxicity in C.elegansAlzheimer’s model[J].CNS NeurolDisord Drug Targets,2015,14(2):295-302.

Application of transgenic Caenorhabditis elegans in Alzheimer’s disease

LI Fan,LUO Jian-ming,LIU Shao-li,HAN Lei,LU Jiang,ZHANG Ya-li*

(College of Food Science and Nutritional Engineering,China Agriculture University,Beijing 100083,China)

Since the Caenorhabditis elegans genome displayes high homology(40%)to human genome and its functions of neurons and neurotransmitters are also highly similar with those of humans,Caenorhabditis elegans have become an effective model of the neural mechanisms of disease.Researches of applications of transgenic Caenorhabditis elegans in Alzheimer’s disease were focused on and the applications of natural extracts in Alzheimer’s disease in recent years were summarized.Also some monomers that were applied in transgenic Caenorhabditis elegans were summed up.

Caenorhabditis elegans;Alzheimer’s disease;naturalextracts;monomer

2015-08-27

李梵(1992-),女,硕士,研究方向:葡萄与葡萄酒工程，E-mail:lifan1992@cau.edu.cn。

张雅丽(1975-),女,博士,副教授,研究方向:葡萄与葡萄酒工程，E-mail:zhangyali@cau.edu.cn。

现代农业产业技术体系专项资金资助(CARS-30-yz-2)。

TS201.1

A

1002-0306(2016)07-0361-07

10.13386/j.issn1002-0306.2016.07.062