High-throughput screening for anti-influenza A virus drugs and study of the mechanism of procyanidin on influenza A virus-induced autophagy

J Biomol Screen. 2012 Jun;17(5):605-17. doi: 10.1177/1087057111435236. Epub 2012 Jan 27.

Abstract

In this research, we have established a high-throughput screening (HTS) platform based on the influenza A virus (IAV) vRNA promoter. Using this HTS platform, we selected 35 medicinal plants out of 83 examples of traditional Chinese medicine and found that 7 examples had not been reported. After examining many previous reports, we found that Vaccinium angustifolium Ait., Vitis vinifera L, and Cinnamomum cassia Presl had a common active compound, procyanidin, and then determined the anti-IAV effect of procyanidin and explored its mechanism of action. With a plaque inhibition assay and a time-of-addition experiment, we found that procyanidin could inhibit the IAV replication at several stages of the life cycle. In the Western blot and EGFP-LC3 localization assays, we found that procyanidin could inhibit the accumulation of LC3II and the dot-like aggregation of EGFP-LC3. In the RT-PCR and Western blot assays, we found procyanidin could inhibit the expression of Atg7, Atg5, and Atg12. Finally, by the bimolecular fluorescence complementation-fluorescence resonance energy transfer and co-immunoprecipitation assays, we found that procyanidin could inhibit the formation of the Atg5-Atg12/Atg16 heterotrimer and the dissociation of the beclin1/bcl2 heterodimer. In conclusion, we have established an HTS platform and identified procyanidin as a novel and promising anti-IAV agent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology*
  • Antiviral Agents / toxicity
  • Apoptosis Regulatory Proteins / chemistry
  • Autophagy / drug effects*
  • Biflavonoids / pharmacology*
  • Biflavonoids / toxicity
  • Catechin / pharmacology*
  • Catechin / toxicity
  • Cell Line
  • Drug Discovery / methods
  • Drugs, Chinese Herbal / pharmacology
  • Drugs, Chinese Herbal / toxicity
  • Gene Expression / drug effects
  • Genes, Reporter
  • High-Throughput Screening Assays / methods*
  • Humans
  • Influenza A virus / drug effects*
  • Influenza A virus / genetics
  • Proanthocyanidins / pharmacology*
  • Proanthocyanidins / toxicity
  • Promoter Regions, Genetic / drug effects
  • Protein Multimerization / drug effects
  • Ubiquitin-Activating Enzymes / chemistry
  • Ubiquitin-Activating Enzymes / genetics
  • Virus Replication / drug effects

Substances

  • Antiviral Agents
  • Apoptosis Regulatory Proteins
  • Biflavonoids
  • Drugs, Chinese Herbal
  • Proanthocyanidins
  • procyanidin
  • Catechin
  • Ubiquitin-Activating Enzymes