Acetylharpagide mitigates the detrimental impact of the duck tembusu virus on BHK-21 cells through the inhibition of endoplasmic reticulum apoptosis pathway.
TL;DR
These findings firstly demonstrate the antiviral activities of AHPG against DTMUV, suggesting AHPG possesses mild anti-DTMUV activity and prominent cytoprotective effects against DTMUV-induced cell injury, which renders it a promising cytoprotective candidate for alleviating DTMUV-triggered cellular damage.
Abstract
The duck tembusu virus (DTMUV) is a newly emerging avian pathogenic flavivirus characterized by nonsuppurative encephalitis and reducing egg production that has caused enormous economic loss to the poultry industry in China and surrounding countries. At present, there are few studies on drugs that can protect against DTMUV infection. Acetylharpagide (AHPG) is one of active ingredients of traditional Chinese medicine that plays a role in various biological functions. However, it is not known whether AHPG can inhibit DTMUV infection. Therefore, this study investigated the anti-DTMUV effects and underlying mechanisms of AHPG in vitro. The safe concentration range of AHPG in BHK-21 cells was determined using the MTT assay. Antiviral activity and mechanisms were further evaluated by virus titration, reactive oxygen species (ROS) measurement, Caspase-3 activity assay, mitochondrial membrane potential (MMP) detection, flow cytometry, quantitative real-time PCR, and western blot analysis. Our results showed that AHPG effectively alleviated endoplasmic reticulum stress (ERS) and attenuated DTMUV-induced apoptosis in BHK-21 cells. AHPG significantly inhibited DTMUV-induced oxidative stress, stabilized mitochondrial membrane potential, and reduced cellular apoptosis. Mechanistically, AHPG down-regulated the expression of PERK, ATF4, CHOP and GRP78, up-regulated the anti-apoptotic protein Bcl-2, and down-regulated the pro-apoptotic protein Bax, thereby suppressing the endoplasmic reticulum apoptosis pathway and exerting a protective effect. Taken together, these findings firstly demonstrate the antiviral activities of AHPG against DTMUV, suggesting AHPG possesses mild anti-DTMUV activity and prominent cytoprotective effects against DTMUV-induced cell injury, which renders it a promising cytoprotective candidate for alleviating DTMUV-triggered cellular damage.