title: High-throughput screening for anti-influenza A virus drugs and study of the mechanism of procyanidin on influenza A virus-induced autophagy.
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.
authors.affiliation: Shantou University Medical College, Shantou, Guangdong, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Dai
authors.forename: Gefei
authors.initials: G
authors.lastname: Wang
authors.forename: Weizhong
authors.initials: W
authors.lastname: Li
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Jiacai
authors.initials: J
authors.lastname: Yang
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Zhao
authors.forename: Xiaoxuan
authors.initials: X
authors.lastname: Chen
authors.forename: Yanxuan
authors.initials: Y
authors.lastname: Xu
authors.forename: Kangsheng
authors.initials: K
authors.lastname: Li
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology* / Q000633:toxicity; D051017:Apoptosis Regulatory Proteins / Q000737:chemistry; D001343:Autophagy / Q000187:drug effects*; D044946:Biflavonoids / Q000494:pharmacology* / Q000633:toxicity; D002392:Catechin / Q000494:pharmacology* / Q000633:toxicity; D002460:Cell Line; D055808:Drug Discovery / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000633:toxicity; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects* / Q000235:genetics; D044945:Proanthocyanidins / Q000494:pharmacology* / Q000633:toxicity; D011401:Promoter Regions, Genetic / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D044764:Ubiquitin-Activating Enzymes / Q000737:chemistry / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D051017:Apoptosis Regulatory Proteins; D044946:Biflavonoids; D004365:Drugs, Chinese Herbal; D044945:Proanthocyanidins; C017674:procyanidin; D002392:Catechin; D044764:Ubiquitin-Activating Enzymes