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Additive Effects of Water-Soluble Propolis(Greit 120)and Human Interferon Alfa(HuIFN-αN3)against Influenza Viruses in Vitro

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Journal of Agricultural Science and Technology(A) 2018年第1期8卷 36-47页

作者： Bratko Filipic Lidija Gradisnik Adriana Pereyra Rok Kopino Klemen Rihar Eva Ruzic-Sabljic Snezana Kramar Damir Dermie Eugen Soos nicola volpi Alfredo Fachini and Hrvoje Mazija Croatian Institute of Experimental and Translation Oncology Koledinecka 3Zagreb 10040Croatia Institute of Biomedical Sciences Medical Faculty of MariborTaborska 8Maribor 2000Slovenia Medex D.o.o. Linhartova 49bLjubljana 1000Slovenia Chengdujska 4 Ljubljana 1000Slovenia Institute of Microbiology and Immunology Medical FacultyZaloska 4Ljubljana 1105Slovenia Division of Molecular Biology Ruaer Boskovic InstituteBijenicka 54Zagreb 10000Croatia Trg Sv.Ivana 5 Klostar Ivanic 10312Croatia Department of Life Sciences University of Modena&Reggio EmiliaModena 41125Italy BNatural Via Gran Sasso 33Corbetta-MI 20011Italy

Influenza virus affects the respiratory tract in humans causing a range of distinct manifestations including fever, nasal secretions, cough, headaches, muscle pain and pneumonia, which could become violent and severe. It was found that influenza A viruses remain resistant to amantadine and rimantadin with high level of oseltamvir resistance. Therefore, there is a need for constant improvement of drugs active against resistant influenza viruses. Propolis has anti-influenza activity both in vitro and in vivo. Human leukocyte interferon （HuIFN-αN3） is a multi-subtype protein that displays an antiviral activity against influenza virus. In this study we elucidated the anti-influenza activity of the mixes of water-soluble propolis （WSP）（Greit 120） and HuIFN-αN3 at different ratios. Greit 120 polyphenols, total phenol acids and bioflavonoid were characterized by HPLC-UV-ESI-MS504971 and HuIFN-αN3 by reverse-phase high-performance liquid chromatography （RP-HPLC）. Influenza A and B viruses were separately added to the LLC-MK2 cells treated with WSP （Greit 120） and HuIFN-αN3 alone or in proportions 1：1, 1：2 and 2：1. Plates were incubated and cytopathic effect was determined. The best results （ID50） were obtained with the mix of 10% WSP and HuIFN-αN3 in proportion 1：2, showing ID50 at 12 ± 0.2 μg/mL and 19 ± 0.6 μg/mL for influenza A and B viruses, respectively. When comparing anti-influenza activity of WSP （Greit 120）/HuIFN-αN3 with that of ribavirin, it was found that 1：2 was the optimal ratio for WSP （Greit 120）/HuIFN-αN3 （0.5 and 0.6 for influenza A and B, respectively）. This new formulation of WSP （Greit 120） and HuIFN-αN3, showing better anti-Influenza activity, will definitely improve its application in flu infections.

关键词： Water-soluble propolis Greit 120 HuIFN-αN3 anti-influenza activity ribavirin ID_(50)

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