Inhibition of Porcine Epidemic Diarrhea Virus Replication and Viral 3C-Like Protease by Quercetin
Abstract
:1. Introduction
2. Results
2.1. Toxicity of Quercetin on Cells
2.2. Quercetin Exhibits Antiviral Activity on PEDV
2.3. Quercetin Could not Inhibit PEDV from Attaching to Cellular Membrane and Penetrating Cells
2.4. Quercetin’s Anti-PEDV Effect is Independent of Its Hsp70 Inhibiting Activity
2.5. Potential Sites of PEDV 3CLpro Binding Quercetin
2.6. Quercetin Showed Binding Affinity to PEDV 3CLpro
2.7. Quercetin Inhibits PEDV 3CLpro Activity
3. Discussion
4. Materials and Methods
4.1. Virus, Cells, Antibodies, and Plasmid
4.2. Cytotoxicity Assay
4.3. Quantitative Real-Time RT-PCR
4.4. TCID50 Assay
4.5. Indirect Immunofluorescence Assay
4.6. Western Blot
4.7. Time-Of-Addition Assays
4.8. RNA Interference
4.9. Docking of PEDV 3CLpro and Quercetin
4.10. Protein Expression and Purification
4.11. Surface Plasmon Resonance
4.12. Fluorescence Resonance Energy Transfer Assays for Enzymatic Characteristics
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Pensaert, M.B.; Martelli, P. Porcine epidemic diarrhea: A retrospect from Europe and matters of debate. Virus Res. 2016, 226, 1–6. [Google Scholar] [CrossRef]
- Choudhury, B.; Dastjerdi, A.; Doyle, N.; Frossard, J.-P.; Steinbach, F. From the field to the lab—An European view on the global spread of PEDV. Virus Res. 2016, 226, 40–49. [Google Scholar] [CrossRef] [PubMed]
- Jung, K.; Saif, L.J. Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis. Veter. J. 2015, 204, 134–143. [Google Scholar] [CrossRef]
- Lee, S.; Lee, C. Outbreak-related porcine epidemic diarrhea virus strains similar to US strains, South Korea, 2013. Emerg. Infect. Dis. 2014, 20, 1223–1226. [Google Scholar] [CrossRef] [PubMed]
- Li, W.; Li, H.; Liu, Y.; Pan, Y.; Deng, F.; Song, Y.; Tang, X.; He, Q. New variants of porcine epidemic diarrhea virus, China, 2011. Emerg. Infect. Dis. 2012, 18, 1350–1353. [Google Scholar] [CrossRef] [PubMed]
- Vlasova, A.N.; Al, A.N.V.E.; Wang, Q.; Gramer, M.R.; Rossow, K.D.; Rovira, A.; Collins, J.E.; Saif, L.J. Distinct characteristics and complex evolution of PEDV strains, North America, May 2013–February 2014. Emerg. Infect. Dis. 2014, 20, 1620–1628. [Google Scholar] [CrossRef] [Green Version]
- Harwood, M.; Danielewska-Nikiel, B.; Borzelleca, J.F.; Flamm, G.W.; Williams, G.M.; Lines, T.C. A critical review of the data related to the safety of quercetin and lack of evidence of in vivo toxicity, including lack of genotoxic/carcinogenic properties. Food Chem. Toxicol. 2007, 45, 2179. [Google Scholar] [CrossRef]
- Robaszkiewicz, A.; Balcerczyk, A.; Bartosz, G. Antioxidative and prooxidative effects of quercetin on A549 cells. Cell Biol. Int. 2007, 31, 1245–1250. [Google Scholar] [CrossRef]
- Dajas, F. Life or death: Neuroprotective and anticancer effects of quercetin. J. Ethnopharmacol. 2012, 143, 383–396. [Google Scholar] [CrossRef]
- Li, M.; Xu, Z. Quercetin in a lotus leaves extract may be responsible for antibacterial activity. Arch. Pharmacal Res. 2008, 31, 640–644. [Google Scholar] [CrossRef]
- Choi, H.J.; Song, J.H.; Park, K.S.; Kwon, D.H. Inhibitory effects of quercetin 3-rhamnoside on influenza A virus replication. Eur. J. Pharm. Sci. 2009, 37, 329–333. [Google Scholar] [CrossRef] [PubMed]
- Gao, J.; Xiao, S.; Liu, X.-H.; Wang, L.; Ji, Q.; Mo, D.; Chen, Y. Inhibition of HSP70 reduces porcine reproductive and respiratory syndrome virus replication in vitro. BMC Microbiol. 2014, 14, 64. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liu, J.; Bai, J.; Zhang, L.; Jiang, Z.; Wang, X.; Li, Y.; Jiang, P. Hsp70 positively regulates porcine circovirus type 2 replication in vitro. Virology 2013, 447, 52–62. [Google Scholar] [CrossRef] [Green Version]
- Wu, W.; Li, R.; Li, X.; He, J.; Jiang, S.; Liu, S.; Yang, J. Quercetin as an antiviral agent inhibits influenza A virus (IAV) entry. Viruses 2015, 8, 6. [Google Scholar] [CrossRef]
- Lulu, S.S.; Thabitha, A.; Vino, S.; Priya, A.M.; Rout, M. Naringenin and quercetin—Potential anti-HCV agents for NS2 protease targets. Nat. Prod. Res. 2015, 30, 1–5. [Google Scholar]
- Bachmetov, L.; Gal-Tanamy, M.; Shapira, A.; Vorobeychik, M.; Giterman-Galam, T.; Sathiyamoorthy, P.; Golan-Goldhirsh, A.; Benhar, I.; Tur-Kaspa, R.; Zemel, R. Suppression of hepatitis C virus by the flavonoid quercetin is mediated by inhibition of NS3 protease activity. J. Viral Hepat. 2011, 19, e81–e88. [Google Scholar] [CrossRef] [PubMed]
- Song, D.; Park, B. Porcine epidemic diarrhoea virus: A comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes 2012, 44, 167–175. [Google Scholar] [CrossRef] [PubMed]
- Shi, Y.; Lei, Y.; Ye, G.; Sun, L.; Fang, L.; Xiao, S.; Fu, Z.F.; Yingying, L.; Song, Y.; Peng, G. Identification of two antiviral inhibitors targeting 3C-like serine/3C-like protease of porcine reproductive and respiratory syndrome virus and porcine epidemic diarrhea virus. Veter. Microbiol. 2018, 213, 114–122. [Google Scholar] [CrossRef]
- Gorbalenya, A.E.; Enjuanes, L.; Ziebuhr, J.; Snijder, E.J. Nidovirales: Evolving the largest RNA virus genome. Virus Res. 2006, 117, 17–37. [Google Scholar] [CrossRef]
- Ye, G.; Deng, F.; Shen, Z.; Luo, R.; Zhao, L.; Xiao, S.; Fu, Z.F.; Peng, G. Structural basis for the dimerization and substrate recognition specificity of porcine epidemic diarrhea virus 3C-like protease. Virology 2016, 494, 225–235. [Google Scholar] [CrossRef]
- Ryu, Y.B.; Jeong, H.J.; Kim, J.H.; Kim, Y.M.; Park, J.-Y.; Kim, D.; Naguyen, T.T.H.; Park, S.-J.; Chang, J.S.; Park, K.H. Biflavonoids from Torreya nucifera displaying SARS-CoV 3CLpro inhibition. Bioorganic Med. Chem. 2010, 18, 7940–7947. [Google Scholar] [CrossRef] [PubMed]
- Park, J.-Y.; Yuk, H.J.; Ryu, H.W.; Lim, S.H.; Kim, K.S.; Park, K.H.; Ryu, Y.B.; Lee, W.S. Evaluation of polyphenols from Broussonetia papyrifera as coronavirus protease inhibitors. J. Enzym. Inhib. Med. Chem. 2017, 32, 504–512. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gonzalez, O.; Fontanes, V.; Raychaudhuri, S.; Loo, R.; Loo, J.; Arumugaswami, V.; Sun, R.; Dasgupta, A.; French, S.W. The heat shock protein inhibitor Quercetin attenuates hepatitis C virus production. Hepatology 2009, 50, 1756–1764. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zanini, C.; Giribaldi, G.; Mandili, G.; Carta, F.; Crescenzio, N.; Bisaro, B.; Doria, A.; Foglia, L.; Di Montezemolo, L.C.; Timeus, F.; et al. Inhibition of heat shock proteins (HSP) expression by quercetin and differential doxorubicin sensitization in neuroblastoma and Ewing’s sarcoma cell lines. J. Neurochem. 2007, 103, 1344–1354. [Google Scholar] [CrossRef]
- Lin, C.-W.; Tsai, F.-J.; Tsai, C.-H.; Lai, C.-C.; Wan, L.; Ho, T.-Y.; Hsieh, C.-C.; Chao, P.-D.L. Anti-SARS coronavirus 3C-like protease effects of Isatis indigotica root and plant-derived phenolic compounds. Antivir. Res. 2005, 68, 36–42. [Google Scholar] [CrossRef]
- Nguyen, T.T.H.; Woo, H.-J.; Kang, H.-K.; Nguyen, V.D.; Kim, Y.-M.; Kim, D.-W.; Ahn, S.-A.; Xia, Y.; Kim, D. Flavonoid-mediated inhibition of SARS coronavirus 3C-like protease expressed in Pichia pastoris. Biotechnol. Lett. 2012, 34, 831–838. [Google Scholar] [CrossRef] [Green Version]
- Wang, L.; Bao, B.-B.; Song, G.-Q.; Chen, C.; Zhang, X.-M.; Lu, W.; Wang, Z.; Cai, Y.; Li, S.; Fu-Hang, S.; et al. Discovery of unsymmetrical aromatic disulfides as novel inhibitors of SARS-CoV main protease: Chemical synthesis, biological evaluation, molecular docking and 3D-QSAR study. Eur. J. Med. Chem. 2017, 137, 450–461. [Google Scholar] [CrossRef]
- Xu, H.-X.; Wan, M.; Dong, H.; But, P.P.-H.; Foo, L.Y. Inhibitory activity of flavonoids and tannins against HIV-1 protease. Biol. Pharm. Bull. 2000, 23, 1072–1076. [Google Scholar] [CrossRef] [Green Version]
- Senthilvel, P.; Lavanya, P.; Kumar, K.M.; Swetha, R.; Anitha, P.; Bag, S.; Sarveswari, S.; Vijayakumar, V.; Ramaiah, S.; Anbarasu, A. Flavonoid from Carica papaya inhibits NS2B-NS3 protease and prevents Dengue 2 viral assembly. Bioinformation 2013, 9, 889–895. [Google Scholar] [CrossRef]
- Kakiuchi, N.; Nishikawa, S.; Hattori, M.; Shimotohno, K. A high throughput assay of the hepatitis C virus nonstructural protein 3 serine proteinase. J. Virol. Methods 1999, 80, 77–84. [Google Scholar] [CrossRef]
- Wang, D.; Fang, L.; Shi, Y.; Zhang, H.; Gao, L.; Peng, G.; Chen, H.; Li, K.; Xiao, S. Porcine epidemic diarrhoea virus 3C-like protease regulates its interferon antagonism by cleaving NEMO. J. Virol. 2015, 90, 2090–2101. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ramajayam, R.; Tan, K.-P.; Liu, H.-G.; Liang, P.-H. Synthesis and evaluation of pyrazolone compounds as SARS-coronavirus 3C-like protease inhibitors. Bioorgan. Med. Chem. 2010, 18, 7849–7854. [Google Scholar] [CrossRef]
- Chen, C.-N.; Lin, C.P.C.; Huang, K.-K.; Chen, W.-C.; Hsieh, H.-P.; Liang, P.-H.; Hsu, J.T.-A. Inhibition of SARS-CoV 3C-like Protease Activity by Theaflavin-3,3′-digallate (TF3). Evid. Based Complement. Altern. Med. 2005, 2, 209–215. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, L.; Li, J.; Luo, C.; Liu, H.; Xu, W.; Chen, G.; Liew, O.W.; Zhu, W.; Puah, C.M.; Shen, X. Binding interaction of quercetin-3-β-galactoside and its synthetic derivatives with SARS-CoV 3CL pro: Structure–activity relationship studies reveal salient pharmacophore features. Bioorgan. Med. Chem. 2006, 14, 8295–8306. [Google Scholar] [CrossRef]
- McGurn, L.D.; Moazami-Goudarzi, M.; White, A.; Suwal, T.; Brar, B.; Tang, J.Q.; Espie, G.S.; Kimber, M.S. The structure, kinetics and interactions of the β-carboxysomal β-carbonic anhydrase, CcaA. Biochem. J. 2016, 473, 4559–4572. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Li, Z.; Cao, H.; Cheng, Y.; Zhang, X.; Zeng, W.; Sun, Y.; Chen, S.; He, Q.; Han, H. Inhibition of Porcine Epidemic Diarrhea Virus Replication and Viral 3C-Like Protease by Quercetin. Int. J. Mol. Sci. 2020, 21, 8095. https://doi.org/10.3390/ijms21218095
Li Z, Cao H, Cheng Y, Zhang X, Zeng W, Sun Y, Chen S, He Q, Han H. Inhibition of Porcine Epidemic Diarrhea Virus Replication and Viral 3C-Like Protease by Quercetin. International Journal of Molecular Sciences. 2020; 21(21):8095. https://doi.org/10.3390/ijms21218095
Chicago/Turabian StyleLi, Zhonghua, Hua Cao, Yufang Cheng, Xiaoqian Zhang, Wei Zeng, Yumei Sun, Shuhua Chen, Qigai He, and Heyou Han. 2020. "Inhibition of Porcine Epidemic Diarrhea Virus Replication and Viral 3C-Like Protease by Quercetin" International Journal of Molecular Sciences 21, no. 21: 8095. https://doi.org/10.3390/ijms21218095