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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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Propolis Extracts Enhance the Antimicrobial Activity of Slovenian:Chestnut,Forest and Flower Honeys

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Journal of Agricultural Science and Technology(B) 2019年第3期9卷 203-219页

作者： Bratko Filipic Klemen Rihar Dunja Exel Gregoric Lidija Gradisnik Adriana Pereyra Eva Ruzic-Sabljic Fabio Galeotti alfredo fachini Hrvoje Mazija Croatian Institute for Experimental and Translational Oncology(CIETO) Koledinecka 03Zagreb 10040Croatia Retired Chengdujska 4Ljubljana 1000Slovenia Retired Glavarjeva 19Ljubljana 1000Slovenia Institute of Biomedical Sciences Medical Faculty of MariborTaborska 8Maribor 2000Slovenia Medex D.o.o. Linhartova 49aLjubljana 1000Slovenia Institute for Microbiology and Immunology Medical Faculty of LjubljanaZloska 04Ljubljana 1105Slovenia Department of Life Sciences University of Modena&Regio EmiliaModena 41125Italy BNatural Via Gran SassoCorbettaMI 20011Italy

Because of the overuse and misuse of antibiotic, an increase in antibiotic resistance of pathogenic bacteria is evolving. Attention should be focused on natural alternatives to antibiotics, like propolis and honey. They all have strong antibacterial properties because of the active substances they contain. In the present study, the influence of P1 (water-soluble propolis (WSP) Greit 120) or P2 (phosphate buffer saline (PBS) propolis extract Lj. Center pH = 8.0) on antimicrobial activity of different Slovenian: chestnut honey (CH), forest honeys (FoH), and flower honey (FlH) against various Gram-positive or Gram-negative bacteria and yeast Candida albicans are shown. Gram-positive bacteria used in the experiments were: methicillin-resistant Staphylococcus aureus, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus mutans. The yeast was Candia albicans. Gram-negative bacteria used were: Escherichia coli, Pseudomonas aeruginosa and Acinetobacter baumanii. P1, P2, FoH, CH, FlH and their combinations were prepared in saline as 10% solutions. The composition of both propolis extracts and the three kinds of honey was determined by reverse-phase high-performance liquid chromatography (RP-HPLC). It expressed the antimicrobial activity as a minimal inhibitory concentration (MIC) in mg/mL. Each of the two propolis extracts showed antimicrobial activity when applied alone. Also, the application of each of honey yielded an antimicrobial activity in decreasing order: CH, FoH and FlH honey. The combination of any of the propolis extract with any of the honey showed the additive antimicrobial effect on either Gram-positive or Gram-negative bacteria and on yeast Candida albicans. The effectiveness of P1 or P2 depends on their nature and from the sort of honey and is the highest in the case of CH followed by FoH and FlH.

关键词： Honey propolis flavonoids antimicrobial activity enhancement

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