盐霉素体外对猪流行性腹泻病毒的抑制效果

doi:10.11843/j.issn.0366-6964.2024.04.028

Abstract

Abstract: Porcine epidemic diarrhea (PED) is a porcine intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV), characterized by watery diarrhea, vomiting and dehydration. It has caused huge economic losses to the global pig industry, but there are still no effective vaccines and treatments. Salinomycin (SLM) is a commonly used antibiotic with the advantages of low resistance, rapid excretion and very low residue. To explore the inhibitory effect of SLM on PEDV, firstly, the proliferation pattern of PEDV on Vero cells was detected by TCID₅₀, and then the CC₅₀ and IC₅₀ of SLM were measured by CCK-8 test and cytopathic effect (CPE). Finally, the co-culture models of SLM, PEDV and Vero cell were established. The effects of SLM on the replication cycle of PEDV were determined by RT-qPCR, Western blot and indirect immunofluorescent assay. The results showed that the highest virus titer (10^7.7·0.1 mL^-1) was observed in Vero cells at 24 hours after PEDV infection. The CC₅₀ of SLM on Vero cells was 7.698 μmol·L^-1, and the IC₅₀ for PEDV was 1.617 μmol·L^-1. The study showed that viral titer, N protein and N gene mRNA expression of PEDV gradually decreased with the increase of SLM concentration. SLM mainly inhibited the replication phase of PEDV, and had no significant effect on the adsorption, invasion and release phases of the virus. The research results could provide new ideas for further exploring PEDV inhibitory drugs.

Key words: PEDV CV777, Vero cell, salinomycin, replication cycle

CLC Number:

R978.7

MA Yajuan, SU Kai, LIN Yidan, WANG Yawen, ZHANG Yanan, YUAN Hongxing, YUAN Chen, SONG Qinye. The Inhibitory Effect of Salinomycin on Porcine Epidemic Diarrhea Virus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1661-1671.

References

[1] KUANG Y Q, MO M J, HE H L, et al. Preparation of monoclonal antibody against N protein of porcine epidemic diarrhea virus and establishment of indirect immuno-fluorescence assay[J]. Journal of South China Agricultural University, 2020, 41(5):27-35. (in Chinese)
邝燕齐, 莫梅君, 何红玲, 等. PEDV N蛋白单克隆抗体的制备及间接免疫荧光检测方法的建立[J]. 华南农业大学学报, 2020, 41(5):27-35.
[2] STADLER K, MASIGNANI V, EICKMANN M, et al. SARS-beginning to understand a new virus[J]. Nat Rev Microbiol, 2003, 1(3):209-218.
[3] CHI S S, CHEN S, JIA W J, et al. Non-structural proteins of bovine viral diarrhea virus[J]. Virus Genes, 2022, 58(6):491-500.
[4] LEE C. Porcine epidemic diarrhea virus:an emerging and re-emerging epizootic swine virus[J]. Virol J, 2015, 12:193.
[5] WEI X N, SHE G L, WU T T, et al. PEDV enters cells through clathrin-, caveolae-, and lipid raft-mediated endocytosis and traffics via the endo-/lysosome pathway[J]. Vet Res, 2020, 51(1):10.
[6] PERLMAN S, NETLAND J. Coronaviruses post-SARS:update on replication and pathogenesis[J]. Nat Rev Microbiol, 2009, 7(6):439-450.
[7] ANTOSZCZAK M, HUCZYŃSKI A. Salinomycin and its derivatives-A new class of multiple-targeted "magic bullets"[J]. Eur J Med Chem, 2019, 176:208-227.
[8] SUN P, BI F F, HAO Z K, et al. Application of salinomycin in the prevention and control of chicken coccidiosis:overview[J]. Journal of Parasites and Medical Entomology, 2021:1-8. (in Chinese)
孙霈, 毕菲菲, 郝振凯, 等. 盐霉素在鸡球虫病防控中的应用:概述[J]. 寄生虫与医学昆虫学报, 2021:1-8.
[9] LI Y S, LIU B X, SUN Q, et al. Inhibition of anti-cancer stem cells drug salinomycin on Wnt/β-catenin signaling pathway[J]. Scientia Sinica Vitae, 2022, 52(2):223-236. (in Chinese)
黎莹斯, 刘博昕, 孙琦, 等. 抗肿瘤干细胞药物盐霉素对Wnt/β-catenin信号通路的抑制[J]. 中国科学:生命科学, 2022, 52(2):223-236.
[10] HUCZYNSKI A. Salinomycin:a new cancer drug candidate[J]. Chem Biol Drug Des, 2012, 79(3):235-238.
[11] CHAPMAN H D, JEFFERS T K, WILLIAMS R B. Forty years of monensin for the control of coccidiosis in poultry[J]. Poult Sci, 2010, 89(9):1788-1801.
[12] KEVIN II D A, MEUJO D A, HAMANN M T. Polyether ionophores:broad-spectrum and promising biologically active molecules for the control of drug-resistant bacteria and parasites[J]. Expert Opin Drug Discov, 2009, 4(2):109-146.
[13] ANTOSZCZAK M. A medicinal chemistry perspective on salinomycin as a potent anticancer and anti-CSCs agent[J]. Eur J Med Chem, 2019, 164:366-377.
[14] PARAJULI B, SHIN S J, KWON S H, et al. Salinomycin induces apoptosis via death receptor-5 up-regulation in cisplatin-resistant ovarian cancer cells[J]. Anticancer Res, 2013, 33(4):1457-1462.
[15] HUANG X L, BORGSTRÖM B, STEGMAYR J, et al. The molecular basis for inhibition of stemlike cancer cells by salinomycin[J]. ACS Cent Sci, 2018, 4(6):760-767.
[16] MAI T T, HAMAÏ A, HIENZSCH A, et al. Salinomycin kills cancer stem cells by sequestering iron in lysosomes[J]. Nat Chem, 2017, 9(10):1025-1033.
[17] HUCZYŃSKI A, ANTOSZCZAK M, KLECZEWSKA N, et al. Synthesis and biological activity of salinomycin conjugates with floxuridine[J]. Eur J Med Chem, 2015, 93:33-41.
[18] PARAJULI B, LEE H G, KWON S H, et al. Salinomycin inhibits Akt/NF-κB and induces apoptosis in cisplatin resistant ovarian cancer cells[J]. Cancer Epidemiol, 2013, 37(4):512-517.
[19] CHEN N H, LI S J, ZHOU R Y, et al. Two novel porcine epidemic diarrhea virus (PEDV) recombinants from a natural recombinant and distinct subtypes of PEDV variants[J]. Virus Res, 2017, 242:90-95.
[20] YAN Q Y, ZHOU S T, XIE Y Y, et al. Cytopathology study on porcine epidemic diarrhea virus (PEDV) infected Vero cells[J]. Journal of Shanghai Jiaotong University (Agricultural Science), 2018, 36(2):39-42. (in Chinese)
闫清源, 周书亭, 谢洋洋, 等. 猪流行性腹泻病毒感染Vero细胞的细胞病理学研究[J]. 上海交通大学学报(农业科学版), 2018, 36(2):39-42.
[21] LI Z H, CAO H, CHENG Y F, et al. Inhibition of porcine epidemic diarrhea virus replication and viral 3C-like protease by quercetin[J]. Int J Mol Sci, 2020, 21(21):8095.
[22] GAO R Y, ZHANG Y S, KANG Y H, et al. Glycyrrhizin inhibits PEDV infection and proinflammatory cytokine secretion via the HMGB1/TLR4-MAPK p38 pathway[J]. Int J Mol Sci, 2020, 21(8):2961.
[23] REN J P, ZENG W, JIANG C S, et al. Inhibition of porcine epidemic diarrhea virus by cinchonine via inducing cellular autophagy[J]. Front Cell Infect Microbiol, 2022, 12:856711.
[24] SUN P, WANG M G, LI J, et al. Inhibitory effect of Buddlejasaponin IVb on porcine epidemic diarrhea virus in vivo and in vitro[J]. Vet Microbiol, 2022, 272:109516.
[25] HUAN C C, XU W Y, NI B, et al. Epigallocatechin-3-gallate, the main polyphenol in Green Tea, inhibits porcine epidemic diarrhea virus in vitro[J]. Front Pharmacol, 2021, 12:628526.
[26] LI L, YU X M, ZHANG H M, et al. In vitro antiviral activity of Griffithsin against porcine epidemic diarrhea virus[J]. Virus Genes, 2019, 55(2):174-181.
[27] WANG P C, BAI J, LIU X W, et al. Tomatidine inhibits porcine epidemic diarrhea virus replication by targeting 3CL protease[J]. Vet Res, 2020, 51(1):136.
[28] JANG Y, SHIN J S, YOON Y S, et al. Salinomycin inhibits influenza virus infection by disrupting endosomal acidification and viral matrix protein 2 function[J]. J Virol, 2018, 92(24):e01441-18.
[29] FRISCH S M, SCHALLER M, CIEPLY B. Mechanisms that link the oncogenic epithelial-mesenchymal transition to suppression of anoikis[J]. J Cell Sci, 2013, 126(Pt 1):21-29.
[30] WEBSTER M R, WEERARATNA A T. A Wnt-er migration:the confusing role of β-catenin in melanoma metastasis[J]. Sci Signal, 2013, 6(268):pe11.
[31] ZUO Y W. GSK-3 mediated β-catenin regulating the proliferation of PEDV[D]. Beijing:Chinese Academy of Agricultural Sciences, 2018. (in Chinese)
左叶雯. GSK-3介导β-catenin调控猪流行性腹泻病毒增殖[D]. 北京:中国农业科学院, 2018.
[32] ZHU L Q, THUNUGUNTLA P, LIU Y L, et al. The β-catenin signaling pathway stimulates bovine herpesvirus 1 productive infection[J]. Virology, 2017, 500:91-95.
[33] ZHANG Y S, LIU L G, LI F, et al. Salinomycin exerts anticancer effects on PC-3 cells and PC-3-derived cancer stem cells in vitro and in vivo[J]. Biomed Res Int, 2017, 2017:4101653.
[34] RAUSCH K, HACKETT B A, WEINBREN N L, et al. Screening bioactives reveals nanchangmycin as a broad spectrum antiviral active against Zika Virus[J]. Cell Rep, 2017, 18(3):804-815.
[35] SVENNINGSEN E B, THYRSTED J, BLAY-CADANET J, et al. Ionophore antibiotic X-206 is a potent inhibitor of SARS-CoV-2 infection in vitro[J]. Antiviral Res, 2021, 185:104988.
[36] IANEVSKI A, YAO R A, FENSTAD M H, et al. Potential antiviral options against SARS-CoV-2 infection[J]. Viruses, 2020, 12(6):642.
[37] YANG C W, PENG T T, HSU H Y, et al. Repurposing old drugs as antiviral agents for coronaviruses[J]. Biomed J, 2020, 43(4):368-374.
[38] DORNE J L C M, FERNÁNDEZ-CRUZ M L, BERTELSEN U, et al. Risk assessment of coccidostatics during feed cross-contamination:animal and human health aspects[J]. Toxicol Appl Pharmacol, 2013, 270(3):196-208.
[39] CHEN L G. The effect of salinomycin on pig production performance[J]. Contemporary Animal Husbandry, 2017(20):76-77. (in Chinese)
陈利国. 盐霉素对猪生产性能的影响[J]. 当代畜牧, 2017(20):76-77.

The Inhibitory Effect of Salinomycin on Porcine Epidemic Diarrhea Virus in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 5

Recommended Articles

Metrics

Comments

[1]	WU Jinyan, TIAN Hong, MENG Xuelian, HUI Xiaoting, WANG Yaojie, GUAN Yuhua, SHANG Youjun, LIU Yongsheng, ZHANG Zhidong. Establishment of Vero/SLAM Cell Lines with Stable Expression SLAM to Enhanced PPRV Replication Using the Lentiviral Expression System [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(1): 218-226.
[2]	ZHU Lu, YAN Xia, DAI Ke, WANG Zheng-hao, ZHAO Yu-jia, YANG Zhen, WEN Xin-tian, CAO San-jie, HUANG Xiao-bo, WU Rui, ZHAO Qin, WEN Yi-ping. Establishment of Vero and BHK21 Cell Lines with a Stable Expression of Canine Distemper Virus Cell Receptor SLAM and Comparison of Their Effects on the Isolation of Canine Distemper Virus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1212-1221.
[3]	ZHENG Ya-ni, GAO Xiu-ge, LIU Xiao-xiao, TENG Pei, JI Hui, JIANG Shan-xiang. Preliminary Research of Toxic Effects and Mechanisms of Salinomycin on Chicken Primary Cardiomyocytes [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(4): 740-751.
[4]	HUANG Li，XIE Zhi-xun，XIE Li-ji，DENG Xian-wen，XIE Zhi-qin，FAN Qing，LUO Si-si，HUANG Jiao-ling，ZENG Ting-ting，ZHANG Yan-fang，WANG Sheng. Proteomic Analysis of Vero Cells Infected by Avian Reovirus Virulent Strain and Attenuated Vaccine Strain [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(2): 331-339.
[5]	FU Bo;MA Hong;REN Liang;ZHAO Jin-feng;FANG Qing-chang;GUO Zhen-hua;LI Zhong-qiu;LIU Di;. Effect of Co-culture System with Vero Cells on the Development of Porcine Embryos [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(8): 962-966.