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.

Versatile and Synergistic Potential of Eugenol: A Review

“…Eugenol also ameliorated the oxidative stress and inhibited the expressions of autophagic genes. Beclin1-Bcl2 heterodimer, autophagy, and impaired IAV replication suggested that eugenol is the promising inhibitor for autophagy and IAV infection [49].…”

Eugenol Ameliorates Hepatic Steatosis and Fibrosis by Down-Regulating SREBP1 Gene Expression <i>via</i> AMPK-mTOR-p70S6K Signaling Pathway

“…As a functional ingredient, it is included in many dental preparations and it has also been shown to enhance skin permeation of various drugs. 4) In recent years, eugenol has been reported to have antioxidant, 5) anti-inflammatory, 6) anti-viral 7) and anticancer [8][9][10] activities. Although beneficial effects of eugenol on oxidative stress and inflammation have been well-studied, pharmacological effects on metabolic diseases are largely unknown.…”

Anti-H1N1 influenza effects and its possible mechanism of Huanglian Xiangru Decoction

“…In the process of inflammation, lung itself has antioxidant defense system. Under normal conditions, the body possesses a set of free radical scavenging enzymes (superoxide dismutase SOD and glutathione peroxidase GSH-PX), which keeps a equilibrium state at low concentration (Dai et al, 2013); when Table 2 Effects of HXD and ribavirin on H1N1 influenza infected mice.…”