Brevilin A, a Sesquiterpene Lactone, Inhibits the Replication of Influenza A Virus In Vitro and In Vivo
Abstract
1. Introduction
2. Materials and Methods
2.1. Compounds and Reagents
2.2. Cells and Viruses
2.3. Animals
2.4. Plaque Assay and Plaque Reduction Assay
2.5. Immunofluorescence
2.6. Western Blot Assay
2.7. Real-Time Quantitative PCR (RT-qPCR)
2.8. In Vivo Experiments
2.9. Statistical Analysis
3. Results
3.1. Brevilin A Shows a Broad-Spectrum Antiviral Activity against IAV
3.2. Brevilin A Inhibits Progeny Virus Production in Various Virus-To-Cell Ratios
3.3. Brevilin A Is Effective at the Viral Genome Replication and Translation Stage
3.4. Brevilin A Inhibits Influenza Viral RNA Synthesis
3.5. Brevilin A Decreases the Levels of Viral mRNA and Proteins Expressed from the M and NS Segments
3.6. Brevilin A Induces Influenza Viral RNP Aggregation in the Nucleus
3.7. Brevilin A Protects Mice from IAV Pathogenesis
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Petrova, V.N.; Russell, C.A. The evolution of seasonal influenza viruses. Nat. Rev. Microbiol. 2018, 16, 47–60. [Google Scholar] [CrossRef] [PubMed]
- Iuliano, A.D.; Roguski, K.M.; Chang, H.H.; Muscatello, D.J.; Palekar, R.; Tempia, S.; Cohen, C.; Gran, J.M.; Schanzer, D.; Cowling, B.J.; et al. Global Seasonal Influenza-associated Mortality Collaborator, N., Estimates of global seasonal influenza-associated respiratory mortality: A modelling study. Lancet 2018, 391, 1285–1300. [Google Scholar] [CrossRef]
- Paules, C.I.; Sullivan, S.G.; Subbarao, K.; Fauci, A.S. Chasing Seasonal Influenza—The Need for a Universal Influenza Vaccine. N. Engl. J. Med. 2018, 378, 7–9. [Google Scholar] [CrossRef] [PubMed]
- Hu, Y.; Musharrafieh, R.; Ma, C.; Zhang, J.; Smee, D.F.; DeGrado, W.F.; Wang, J. An M2-V27A channel blocker demonstrates potent in vitro and in vivo antiviral activities against amantadine-sensitive and -resistant influenza A viruses. Antivir. Res. 2017, 140, 45–54. [Google Scholar] [CrossRef] [PubMed]
- Hussain, M.; Galvin, H.D.; Haw, T.Y.; Nutsford, A.N.; Husain, M. Drug resistance in influenza A virus: The epidemiology and management. Infect. Drug Resist. 2017, 10, 121–134. [Google Scholar] [CrossRef] [PubMed]
- Hayden, F.G.; Sugaya, N.; Hirotsu, N.; Lee, N.; de Jong, M.D.; Hurt, A.C.; Ishida, T.; Sekino, H.; Yamada, K.; Portsmouth, S.; et al. Baloxavir Marboxil for Uncomplicated Influenza in Adults and Adolescents. N. Engl. J. Med. 2018, 379, 913–923. [Google Scholar] [CrossRef] [PubMed]
- O’Hanlon, R.; Shaw, M.L. Baloxavir marboxil: The new influenza drug on the market. Curr. Opin. Virol. 2019, 35, 14–18. [Google Scholar] [CrossRef] [PubMed]
- Bright, R.A.; Medina, M.J.; Xu, X.; Perez-Oronoz, G.; Wallis, T.R.; Davis, X.M.; Povinelli, L.; Cox, N.J.; Klimov, A.I. Incidence of adamantane resistance among influenza A (H3N2) viruses isolated worldwide from 1994 to 2005: A cause for concern. Lancet 2005, 366, 1175–1181. [Google Scholar] [CrossRef]
- Moscona, A. Global transmission of oseltamivir-resistant influenza. N. Engl. J. Med. 2009, 360, 953–956. [Google Scholar] [CrossRef]
- Taniguchi, K.; Ando, Y.; Nobori, H.; Toba, S.; Noshi, T.; Kobayashi, M.; Kawai, M.; Yoshida, R.; Sato, A.; Shishido, T.; et al. Inhibition of avian-origin influenza A(H7N9) virus by the novel cap-dependent endonuclease inhibitor baloxavir marboxil. Sci. Rep. 2019, 9, 3466. [Google Scholar] [CrossRef]
- Omoto, S.; Speranzini, V.; Hashimoto, T.; Noshi, T.; Yamaguchi, H.; Kawai, M.; Kawaguchi, K.; Uehara, T.; Shishido, T.; Naito, A.; et al. Characterization of influenza virus variants induced by treatment with the endonuclease inhibitor baloxavir marboxil. Sci. Rep. 2018, 8, 9633. [Google Scholar] [CrossRef]
- Chan, C.O.; Jin, D.P.; Dong, N.P.; Chen, S.B.; Mok, D.K. Qualitative and quantitative analysis of chemical constituents of Centipeda minima by HPLC-QTOF-MS & HPLC-DAD. J. Pharm. Biomed. Anal. 2016, 125, 400–407. [Google Scholar] [PubMed]
- Liu, Y.; Chen, X.Q.; Liang, H.X.; Zhang, F.X.; Zhang, B.; Jin, J.; Chen, Y.L.; Cheng, Y.X.; Zhou, G.B. Small compound 6-O-angeloylplenolin induces mitotic arrest and exhibits therapeutic potentials in multiple myeloma. PLoS ONE 2011, 6, e21930. [Google Scholar] [CrossRef] [PubMed]
- Wu, P.; Su, M.X.; Wang, Y.; Wang, G.C.; Ye, W.C.; Chung, H.Y.; Li, J.; Jiang, R.W.; Li, Y.L. Supercritical fluid extraction assisted isolation of sesquiterpene lactones with antiproliferative effects from Centipeda minima. Phytochemistry 2012, 76, 133–140. [Google Scholar] [CrossRef] [PubMed]
- Chen, X.; Du, Y.; Nan, J.; Zhang, X.; Qin, X.; Wang, Y.; Hou, J.; Wang, Q.; Yang, J. Brevilin A, a novel natural product, inhibits janus kinase activity and blocks STAT3 signaling in cancer cells. PLoS ONE 2013, 8, e63697. [Google Scholar] [CrossRef] [PubMed]
- Su, M.; Chung, H.Y.; Li, Y. 6-O-Angeloylenolin induced cell-cycle arrest and apoptosis in human nasopharyngeal cancer cells. Chem. Biol. Interact. 2011, 189, 167–176. [Google Scholar] [CrossRef]
- You, P.; Wu, H.; Deng, M.; Peng, J.; Li, F.; Yang, Y. Brevilin A induces apoptosis and autophagy of colon adenocarcinoma cell CT26 via mitochondrial pathway and PI3K/AKT/mTOR inactivation. Biomed. Pharmacother. 2018, 98, 619–625. [Google Scholar] [CrossRef] [PubMed]
- Taylor, R.S.; Towers, G.H. Antibacterial constituents of the Nepalese medicinal herb, Centipeda minima. Phytochemistry 1998, 47, 631–634. [Google Scholar] [CrossRef]
- Yu, H.W.; Wright, C.W.; Cai, Y.; Yang, S.L.; Phillipson, J.D.; Kirby, G.C.; Warhurst, D.C. Antiprotozoal activities of Centipeda minima. Phytother. Res. 1994, 8, 436–438. [Google Scholar] [CrossRef]
- Zhang, X.; He, J.; Huang, W.; Huang, H.; Zhang, Z.; Wang, J.; Yang, L.; Wang, G.; Wang, Y.; Li, Y. Antiviral Activity of the Sesquiterpene Lactones from Centipeda minima against Influenza A Virus in vitro. Nat. Prod. Commun. 2018, 13, 115–119. [Google Scholar] [CrossRef]
- Nachbagauer, R.; Wohlbold, T.J.; Hirsh, A.; Hai, R.; Sjursen, H.; Palese, P.; Cox, R.J.; Krammer, F. Induction of broadly reactive anti-hemagglutinin stalk antibodies by an H5N1 vaccine in humans. J. Virol. 2014, 88, 13260–13268. [Google Scholar] [CrossRef] [PubMed]
- Wang, T.T.; Tan, G.S.; Hai, R.; Pica, N.; Petersen, E.; Moran, T.M.; Palese, P. Broadly protective monoclonal antibodies against H3 influenza viruses following sequential immunization with different hemagglutinins. PLoS Pathog. 2010, 6, e1000796. [Google Scholar] [CrossRef] [PubMed]
- Shapiro, G.I.; Gurney, T., Jr.; Krug, R.M. Influenza virus gene expression: Control mechanisms at early and late times of infection and nuclear-cytoplasmic transport of virus-specific RNAs. J. Virol. 1987, 61, 764–773. [Google Scholar] [PubMed]
- Scull, M.A.; Rice, C.M. A big role for small RNAs in influenza virus replication. Proc. Natl. Acad. Sci. USA 2010, 107, 11153–11154. [Google Scholar] [CrossRef] [PubMed]
- Vester, D.; Lagoda, A.; Hoffmann, D.; Seitz, C.; Heldt, S.; Bettenbrock, K.; Genzel, Y.; Reichl, U. Real-time RT-qPCR assay for the analysis of human influenza A virus transcription and replication dynamics. J. Virol. Methods 2010, 168, 63–71. [Google Scholar] [CrossRef] [PubMed]
- Te Velthuis, A.J.; Fodor, E. Influenza virus RNA polymerase: Insights into the mechanisms of viral RNA synthesis. Nat. Rev. Microbiol. 2016, 14, 479–493. [Google Scholar] [CrossRef] [PubMed]
- Eisfeld, A.J.; Neumann, G.; Kawaoka, Y. At the centre: Influenza A virus ribonucleoproteins. Nat. Rev. Microbiol. 2015, 13, 28–41. [Google Scholar] [CrossRef] [PubMed]
- Watanabe, K.; Takizawa, N.; Katoh, M.; Hoshida, K.; Kobayashi, N.; Nagata, K. Inhibition of nuclear export of ribonucleoprotein complexes of influenza virus by leptomycin B. Virus Res. 2001, 77, 31–42. [Google Scholar] [CrossRef]
- O’Neill, R.E.; Talon, J.; Palese, P. The influenza virus NEP (NS2 protein) mediates the nuclear export of viral ribonucleoproteins. EMBO J. 1998, 17, 288–296. [Google Scholar] [CrossRef]
- Chiang, C.; Chen, G.W.; Shih, S.R. Mutations at Alternative 5′ Splice Sites of M1 mRNA Negatively Affect Influenza A Virus Viability and Growth Rate. J. Virol. 2008, 82, 10873–10886. [Google Scholar] [CrossRef]
- Hutchinson, E.C.; Curran, M.D.; Read, E.K.; Gog, J.R.; Digard, P. Mutational analysis of cis-acting RNA signals in segment 7 of influenza A virus. J. Virol. 2008, 82, 11869–11879. [Google Scholar] [CrossRef] [PubMed]
- Dubois, J.; Terrier, O.; Rosa-Calatrava, M. Influenza viruses and mRNA splicing: Doing more with less. MBio 2014, 5, e00070-14. [Google Scholar] [CrossRef] [PubMed]
Comp. | IAV | EC50 (µM) a | SI b |
---|---|---|---|
brevilin A | A/PR/8/34 H1N1 | 2.96 ± 1.10 | 8 |
A/FM/1/47 H1N1 | 1.60 ± 1.14 | 14 | |
A/Hong Kong/498/97 H3N2 | 3.28 ± 1.09 | 7 | |
A/chicken/Guangdong/1996 H9N2 | 2.07 ± 1.12 | 11 | |
ribavirin | A/PR/8/34 H1N1 | 7.05 ± 1.10 | >14 |
A/FM/1/47 H1N1 | 9.19 ± 1.02 | >20 | |
A/Hong Kong/498/97 H3N2 | 10.76 ± 1.07 | >18 | |
A/chicken/Guangdong/1996 H9N2 | 10.35 ± 1.04 | >18 |
© 2019 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
Zhang, X.; Xia, Y.; Yang, L.; He, J.; Li, Y.; Xia, C. Brevilin A, a Sesquiterpene Lactone, Inhibits the Replication of Influenza A Virus In Vitro and In Vivo. Viruses 2019, 11, 835. https://doi.org/10.3390/v11090835
Zhang X, Xia Y, Yang L, He J, Li Y, Xia C. Brevilin A, a Sesquiterpene Lactone, Inhibits the Replication of Influenza A Virus In Vitro and In Vivo. Viruses. 2019; 11(9):835. https://doi.org/10.3390/v11090835
Chicago/Turabian StyleZhang, Xiaoli, Yiping Xia, Li Yang, Jun He, Yaolan Li, and Chuan Xia. 2019. "Brevilin A, a Sesquiterpene Lactone, Inhibits the Replication of Influenza A Virus In Vitro and In Vivo" Viruses 11, no. 9: 835. https://doi.org/10.3390/v11090835
APA StyleZhang, X., Xia, Y., Yang, L., He, J., Li, Y., & Xia, C. (2019). Brevilin A, a Sesquiterpene Lactone, Inhibits the Replication of Influenza A Virus In Vitro and In Vivo. Viruses, 11(9), 835. https://doi.org/10.3390/v11090835