抗弓形虫和新孢子虫药物体外筛选方法的研究进展

doi:10.12160/j.issn.1672-5190.2023.01.019

摘要/Abstract

摘要：

弓形虫和新孢子虫是2种在形态结构及生物学特征方面非常相似的细胞内寄生性原虫，二者引起的疾病对人和动物健康造成了严重危害。开发和筛选高效、低毒的抗弓形虫和新孢子虫药物一直是抗原虫药物研究领域的重要方向之一。药物筛选包括体外筛选和体内筛选，体外筛选的结果是进行体内筛选的依据和基础。对抗弓形虫和新孢子虫药物的体外筛选方法研究进展进行综述，以期为开发更加高效的抗弓形虫和新孢子虫药物体外筛选技术提供参考。

关键词: 弓形虫, 新孢子虫, 药物筛选方法, 转基因虫体, qPCR

Abstract:

Toxoplasma gondii （T. gondii） and Neospora caninum （N. caninum） are intracellular parasitic protozoa with very similar morphological structure and biological characteristics. The diseases caused by these parasites pose serious harm to human and animal health. Development and screening of high-efficient and low-toxic drugs against T. gondii and N. caninum has long been one of the important directions in the research field of antiprotozoal drugs. Drug screening includes the experiments in vitro and in vivo, and the foundings of screening in vitro are the basis and foundation for selection in vivo. The recent advances in screening technology of drugs against T. gondii and N. caninum in vitro are reviewed, in hoping to provide references for the development of more efficient screening technology of drugs against these parasites in vitro.

Key words: Toxoplasma gondii, Neospora caninum, drug screening technology, transgenic parasites, qPCR

中图分类号:

刘海燕, 李小瑾, 严文岚, 赵立卓, 王天奇, 钱伟锋. 抗弓形虫和新孢子虫药物体外筛选方法的研究进展[J]. 畜牧与饲料科学, 2023, 44(1): 122-128.

LIU Hai-yan, LI Xiao-jin, YAN Wen-lan, ZHAO Li-zhuo, WANG Tian-qi, QIAN Wei-feng. Recent Advances in Screening Technology of Drugs against Toxoplasma gondii and Neospora caninum In Vitro[J]. Animal Husbandry and Feed Science, 2023, 44(1): 122-128.

图/表 1

参考文献 51

[1]	DUBEY J P, HATTEL A L, LINDSAY D S, et al. Neonatal Neospora caninum infection in dogs: Isolation of the causative agent and experimental transmission[J]. Journal of the American Veterinary Medical Association, 1988, 193(10):1259-1263.
[2]	REICHEL M P, ALEJANDRA AYANEGUI-ALCÉRRECA M, GONDIM L F P, et al. What is the global economic impact of Neospora caninum in cattle - the billion dollar question[J]. International Journal for Parasitology, 2013, 43(2):133-142. doi: 10.1016/j.ijpara.2012.10.022
[3]	王旗, 杨雯, 杨小迪, 等. 抗弓形虫药物的研究进展[J]. 中国病原生物学杂志, 2014, 9(5):478-480.
[4]	梁馨一, 葛冰洁, 赵鹏. 抗新孢子虫病药物研究进展[J]. 吉林畜牧兽医, 2020, 41(11):11-12,14.
[5]	刘杨, 张永浩, 李哲, 等. 蒿甲醚体外抗弓形虫作用的实验研究[J]. 中国寄生虫学与寄生虫病杂志, 1997, 15(6):366-369.
[6]	许丽芳, 杨秋林, 沈元琼, 等. 红霉素等四种抗生素体外抗弓形虫作用的研究[J]. 中国人兽共患病杂志, 2004, 20(10):885-887.
[7]	官剑武. 三种蜘蛛毒素抗弓形虫药物的筛选及其作用机制的初探[D]. 长沙: 中南大学, 2010.
[8]	MAZUZ M L, HAYNES R, SHKAP V, et al. Neospora caninum:in vivo and in vitro treatment with artemisone[J]. Veterinary Parasitology, 2012, 187(1/2):99-104. doi: 10.1016/j.vetpar.2011.12.020
[9]	ANDRADE R M, CHAPARRO J D, CAPPARELLI E, et al. Auranofin is highly efficacious against Toxoplasma gondii in vitro and in an in vivo experimental model of acute toxoplasmosis[J]. PLoS Neglected Tropical Diseases, 2014, 8(7):e2973. doi: 10.1371/journal.pntd.0002973
[10]	ZHANG H, LIU J, YANG C S, et al. Triclosan inhibits the growth of Neospora caninum in vitro and in vivo[J]. Parasitology Research, 2019, 118(10):3001-3010. doi: 10.1007/s00436-019-06449-w
[11]	QIAN W F, WANG H, SHAN D, et al. Activity of several kinds of drugs against Neospora caninum[J]. Parasitology International, 2015, 64(6):597-602. doi: 10.1016/j.parint.2015.08.002
[12]	钱伟锋, 山丹, 郝攀, 等. 几种药物抗弓形虫活性的检测[J]. 中国人兽共患病学报, 2020, 36(11):907-912.
[13]	HUFFMAN A M, AYARIGA J A, NAPIER A, et al. Inhibition of Toxoplasma gondii growth by dihydroquinine and its mechanisms of action[J]. Frontiers in Cellular and Infection Microbiology, 2022, 12:852889. doi: 10.3389/fcimb.2022.852889
[14]	YANG X Y, HUANG B, CHEN J P, et al. In vitro effects of aqueous extracts of Astragalus membranaceus and Scutellaria baicalensis GEORGI on Toxoplasma gondii[J]. Parasitology Research, 2012, 110(6):2221-2227. doi: 10.1007/s00436-011-2752-2
[15]	赵莹, 吕芳丽. 匹多莫德体外抑制弓形虫增殖作用的研究[J]. 热带医学杂志, 2013, 13(5):531-534,623.
[16]	YBAÑEZ R H D, LEESOMBUN A, NISHIMURA M, et al. In vitro and in vivo effects of the phytohormone inhibitor fluridone against Neospora caninum infection[J]. Parasitology International, 2016, 65(4):319-322. doi: 10.1016/j.parint.2016.03.009
[17]	LEESOMBUN A, BOONMASAWAI S, NISHIKAWA Y. Effects of Thai Piperaceae plant extracts on Neospora caninum infection[J]. Parasitology International, 2017, 66(3):219-226. doi: 10.1016/j.parint.2017.01.017
[18]	LEESOMBUN A, BOONMASAWAI S, SHIMODA N, et al. Effects of extracts from Thai Piperaceae plants against infection with Toxoplasma gondii[J]. PLoS One, 2016, 11(5):e0156116. doi: 10.1371/journal.pone.0156116
[19]	谢素珠, 李航, 赵少伟, 等. 不同药物体外抑制新孢子虫的比较[J]. 中国兽医学报, 2019, 39(9):1753-1757.
[20]	GUO H P, GAO Y, N′DA D D, et al. In vitro anti-Toxoplasma gondii efficacy of synthesised benzyltriazole derivatives[J]. The Onderstepoort Journal of Veterinary Research, 2021, 88(1):e1-e8.
[21]	ESPOSITO M, STETTLER R, MOORES S L, et al. In vitro efficacies of nitazoxanide and other thiazolides against Neospora caninum tachyzoites reveal antiparasitic activity independent of the nitro group[J]. Antimicrobial Agents and Chemotherapy, 2005, 49(9):3715-3723. doi: 10.1128/AAC.49.9.3715-3723.2005
[22]	李璐, 王晓岑, 张楠, 等. 荜拨体外抗新孢子虫的作用[J]. 中国生物制品学杂志, 2020, 33(5):516-520.
[23]	梁馨一. 氢溴酸槟榔碱抗新孢子虫作用及其机理的初步研究[D]. 延吉: 延边大学, 2021.
[24]	ZAKI L, GHAFFARIFAR F, SHARIFI Z, et al. Effect of imiquimod on tachyzoites of Toxoplasma gondii and infected macrophages in vitro and in BALB/c mice[J]. Frontiers in Cellular and Infection Microbiology, 2020, 10:387. doi: 10.3389/fcimb.2020.00387
[25]	OKEOMA C M, STOWELL K M, WILLIAMSON N B, et al. Neospora caninum:Quantification of DNA in the blood of naturally infected aborted and pregnant cows using real-time PCR[J]. Experimental Parasitology, 2005, 110(1):48-55. doi: 10.1016/j.exppara.2005.01.008
[26]	GHALMI F, CHINA B, KAIDI R, et al. Detection of Neospora caninum in dog organs using real time PCR systems[J]. Veterinary Parasitology, 2008, 155(1/2):161-167. doi: 10.1016/j.vetpar.2008.04.007
[27]	MCFADDEN D C, SEEBER F, BOOTHROYD J C. Use of Toxoplasma gondii expressing beta-galactosidase for colorimetric assessment of drug activity in vitro[J]. Antimicrobial Agents and Chemotherapy, 1997, 41(9):1849-1853. doi: 10.1128/AAC.41.9.1849
[28]	LIU S X, WU M M, HUA Q Q, et al. Two old drugs,NVP-AEW541 and GSK-J4,repurposed against the Toxoplasma gondii RH strain[J]. Parasites and Vectors, 2020, 13(1):242. doi: 10.1186/s13071-020-04094-2
[29]	PEREIRA L M, MOTA C M, BARONI L, et al. Inhibitory action of phenothiazinium dyes against Neospora caninum[J]. Scientific Reports, 2020, 10:7483. doi: 10.1038/s41598-020-64454-x
[30]	PEREIRA L M, VIGATO-FERREIRA I C, DE LUCA G, et al. Evaluation of methylene blue,pyrimethamine and its combination on an in vitro Neospora caninum model[J]. Parasitology, 2017, 144(6):827-833. doi: 10.1017/S0031182016002584
[31]	PEREIRA L M, DE LUCA G, ABICHABKI N L M, et al. Synergic in vitro combinations of artemisinin,pyrimethamine and methylene blue against Neospora caninum[J]. Veterinary Parasitology, 2018, 249:92-97. doi: 10.1016/j.vetpar.2017.11.014
[32]	PEREIRA L M, DE LUCA G, ABICHABKI N L M, et al. Atovaquone,chloroquine,primaquine,quinine and tetracycline:Antiproliferative effects of relevant antimalarials on Neospora caninum[J]. Brazilian Journal of Veterinary Parasitology, 2021, 30(1):e022120.
[33]	王海琦, 陆方贤, 张建宁. 阿奇霉素抗弓形虫感染的体外实验研究[J]. 中国人兽共患病杂志, 2002, 18(4):94-96.
[34]	吴亮, 陈盛霞, 姜旭淦, 等. 银杏酸、阿奇霉素和大蒜素抗弓形虫增殖效果研究[J]. 中国人兽共患病学报, 2008, 24(8):736-740.
[35]	赵鹏, 张守发, 鞠玉琳, 等. 桦褐孔菌多糖和黄芩苷体外抗新孢子虫试验[J]. 畜牧与兽医, 2016, 48(4):146-148.
[36]	SHOJAEE S, FIROUZEH N, KESHAVARZ H, et al. Nanosilver colloid inhibits Toxoplasma gondii tachyzoites and bradyzoites in vitro[J]. Iranian Journal of Parasitology, 2019, 14(3):362-367.
[37]	赵鹏宇, 徐乐乐, 金春美, 等. 长白瑞香提取物的急性毒性及其体外抗弓形虫的研究[J]. 延边大学学报(自然科学版), 2020, 46(1):65-68.
[38]	刘艳秋, 张可华, 王运良, 等. CCK-8和MTS法检测人羊膜上皮细胞增殖的比较[J]. 中国康复理论与实践, 2012, 18(9):827-830.
[39]	PFEFFERKORN E R, PFEFFERKORN L C. Specific labeling of intracellular Toxoplasma gondii with uracil[J]. The Journal of Protozoology, 1977, 24(3):449-453. doi: 10.1111/jeu.1977.24.issue-3
[40]	YOUN H J, LAKRITZ J, KIM D Y, et al. Anti-protozoal efficacy of medicinal herb extracts against Toxoplasma gondii and Neospora caninum[J]. Veterinary Parasitology, 2003, 116(1):7-14. doi: 10.1016/S0304-4017(03)00154-7
[41]	CHEN S X, WU L, JIANG X G, et al. Anti-Toxoplasma gondii activity of GAS in vitro[J]. Journal of Ethnopharmacology, 2008, 118(3):503-507. doi: 10.1016/j.jep.2008.05.023
[42]	INNES E A, PANTON W R, MARKS J, et al. Interferon gamma inhibits the intracellular multiplication of Neospora caninum,as shown by incorporation of ³H uracil[J]. Journal of Comparative Pathology, 1995, 113(1):95-100. doi: 10.1016/S0021-9975(05)80075-1
[43]	OURA C A, INNES E A, WASTLING J M, et al. The inhibitory effect of ovine recombinant interferon-gamma on intracellular replication of Toxoplasma gondii[J]. Parasite Immunology, 1993, 15(9):535-538. doi: 10.1111/pim.1993.15.issue-9
[44]	THOMAS J A, COLLINS C R, DAS S, et al. Development and application of a simple plaque assay for the human malaria parasite Plasmodium falciparum[J]. PLoS One, 2016, 11(6):e0157873. doi: 10.1371/journal.pone.0157873
[45]	雷涛. 棒状体蛋白18介导弓形虫和新孢子虫毒力差异的分子机制研究[D]. 北京: 中国农业大学, 2014.
[46]	SANTOS J M, FERGUSON D J P, BLACKMAN M J, et al. Intramembrane cleavage of AMA1 triggers Toxoplasma to switch from an invasive to a replicative mode[J]. Science, 2011, 331(6016):473-477. doi: 10.1126/science.1199284
[47]	TANG Y Q, HOU S J, LI X Y, et al. Anti-parasitic effect on Toxoplasma gondii induced by a spider peptide lycosin-Ⅰ[J]. Experimental Parasitology, 2019, 198:17-25. doi: 10.1016/j.exppara.2019.01.009
[48]	潘格. 姜黄素联合青蒿琥酯体内外抗弓形虫作用机制研究[D]. 济南: 山东大学, 2021.
[49]	唐云剑, 高剑, 李莹花, 等. 复方一枝蒿颗粒体外抗弓形虫RH株速殖子的超微结构观察[J]. 中国热带医学, 2018, 18(12):1190-1193. doi: 10.13604/j.cnki.46-1064/r.2018.12.04
[50]	文秋嘉, 李淑红. 胡桃楸提取物抗弓形虫作用的体外实验和超微研究[J]. 临床和实验医学杂志, 2010, 9(6):406-409.
[51]	DARIUS A K, MEHLHORN H, HEYDORN A O. Effects of toltrazuril and ponazuril on the fine structure and multiplication of tachyzoites of the NC-1 strain of Neospora caninum (a synonym of Hammondia heydorni) in cell cultures[J]. Parasitology Research, 2004, 92(6):453-458. doi: 10.1007/s00436-003-1063-7