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Evaluation of the anti-neuraminidase activity of the traditional Chinese medicines and determination of the anti-influenza A virus effects of the neuraminidase inhibitory TCMs in vitro and in vivo
Authors:Tian Li  Wang Zhiyong  Wu Hao  Wang Song  Wang Ye  Wang Yanyan  Xu Jingwei  Wang Liying  Qi Fengchun  Fang Minli  Yu Dahai  Fang Xuexun
Institution:a Key Laboratory of Molecular Enzymology and Enzyme Engineering of Ministry Education, Jilin University, Changchun 130012, PR China
b Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, PR China
c Department of Molecular Biology or Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, PR China
d Changchun Institute of Biological Products, Changchun 130062, PR China
Abstract:

Etnopharmacological relevance

Neuraminidase (NA) inhibitors are currently the most effective drugs to treat influenza A viruses infection. Many traditional Chinese medicines (TCMs) have been used in the clinics to treat influenza. The anti-viral mechanisms of these TCMs and their inhibitory effects towards NA need to be systematically tested.

Aim of the study

To evaluate the anti-NA activity of the TCMs and the anti-influenza A virus effects of the NA inhibitory TCMs in vitro and in vivo.

Material and methods

We tested the inhibitory activity of water extracts from 439 TCMs towards NA. The in vitro anti-influenza virus activities of the 5 TCMs were evaluated using the strain A/California/7/2009 (H1N1) NYMC X-179A of influenza A virus. A randomly selected TCM with NA inhibitory activity, Melia toosendan extract, was further evaluated using a mouse model infected with influenza A virus.

Results

Five TCMs, Duchesnea indica (Andr.) Focke Fragaria indica Andr.], Liquidambar formosana Hance., Lithospermum erythrorhizon Sieb. et Zucc., Melia toosendan Sieb. et Zucc., and Prunella vulgaris L., exerted potent inhibitory activity towards NA. These TCMs in the range of 25-250 μg/mL had the ability to reduce virus-induced cytopathic effect (CPE) and the virus yield in MDCK cells. Melia toosendan significantly reduced death rate and prolonged mean day to death (MDD) of the viral infected mice.

Conclusions

This study describes five TCMs exerted strong inhibitory activities towards NA, and exhibited antiviral effect against influenza A virus by reducing viral reproduction and reduced CPE of the viral infected cells. Melia toosendan, significantly reduced death rate and prolonged survival of the H1N1 viral infected mice.
Keywords:NA  neuraminidase  HA  hemagglutinin  TCID50  50% tissue culture infective dose  IC50  50% inhibitory concentration  CC50  half-maximum cytotoxic concentration  DMEM  Dulbecco's modified Eagle's medium  MTT  3-[4  5-dimethyl-thiazol-2-yl]-2  5-diphenyl tetrazolium bromide  TCMs  traditional Chinese medicines  CPE  cytopathic effect  MES  2-N-morpholino-ethanesulfonic acid  MU-NANA  2&prime  d-N-acetylneuraminic acid" target="_blank">-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid  FBS  fetal bovine serum  MDD  mean day to death  LD50  50% median lethal dose  KLP  Melia toosendan Sieb   et Zucc
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