title: Drug screening for autophagy inhibitors based on the dissociation of Beclin1-Bcl2 complex using BiFC technique and mechanism of eugenol on anti-influenza A virus activity.
abstract: Autophagy is involved in many human diseases, such as cancer, cardiovascular disease and virus infection, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), influenza A virus (IAV) and coxsackievirus B3/B4 (CVB3/B4), so a drug screening model targeting autophagy may be very useful for the therapy of these diseases. In our study, we established a drug screening model based on the inhibition of the dissociation of Beclin1-Bcl2 heterodimer, an important negative regulator of autophagy, using bimolecular fluorescence complementation (BiFC) technique for developing novel autophagy inhibitors and anti-IAV agents. From 86 examples of traditional Chinese medicines, we found Syzygium aromaticum L. had the best activity. We then determined the anti-autophagy and anti-IAV activity of eugenol, the major active compound of Syzygium aromaticum L., and explored its mechanism of action. Eugenol could inhibit autophagy and IAV replication, inhibited the activation of ERK, p38MAPK and IKK/NF-κB signal pathways and antagonized the effects of the activators of these pathways. Eugenol also ameliorated the oxidative stress and inhibited the expressions of autophagic genes. We speculated that the mechanism underlying might be that eugenol inhibited the oxidative stress and the activation of ERK1/2, p38MAPK and IKK/NF-κB pathways, subsequently inhibited the dissociation of Beclin1-Bcl2 heterodimer and autophagy, and finally impaired IAV replication. These results might conversely display the reasonableness of the design of our screening model. In conclusion, we have established a drug screening model for developing novel autophagy inhibitor, and find eugenol as a promising inhibitor for autophagy and IAV infection.
authors.affiliation: Department of Microbiology and Immunology, Shantou University Medical College, Shantou, Guangdong, People's Republic of China.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Dai
authors.forename: Xiang-Feng
authors.initials: XF
authors.lastname: Zhao
authors.forename: Jun
authors.initials: J
authors.lastname: Zeng
authors.forename: Qian-Ying
authors.initials: QY
authors.lastname: Wan
authors.forename: Jia-Cai
authors.initials: JC
authors.lastname: Yang
authors.forename: Wei-Zhong
authors.initials: WZ
authors.lastname: Li
authors.forename: Xiao-Xuan
authors.initials: XX
authors.lastname: Chen
authors.forename: Ge-Fei
authors.initials: GF
authors.lastname: Wang
authors.forename: Kang-Sheng
authors.initials: KS
authors.lastname: Li
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D005054:Eugenol / Q000494:pharmacology*; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects*; D027842:Syzygium / Q000737:chemistry
chemical_list: D000998:Antiviral Agents; D004365:Drugs, Chinese Herbal; D005054:Eugenol