Study on antiviral activity of isochlorogenic acid A against peste des petit ruminant virus in vitro and its action mechanism
Journal Title: Journal of Henan Agricultural University - Year 2023, Vol 57, Issue 1
Abstract
[Objective] The present work was conducted to investigate the antiviral activity of isochlorogenic acid A (IAA) against peste des petits ruminant virus (PPRV) in vitro and its mechanism. [Methods] The maximum non-toxic mass concentration and the 50% cytotoxic mass concentration (CC50) of IAA on African green monkey kidney cells (verda reno, Vero) were determined by MTT method. The median effective mass concentration (EC50) of IAA was determined by plaque formation test and the therapeutic index (TI) was calculated. The effect of IAA on the cell viability of the PPRV-infected Vero cells was determined by MTT assay, and the effects of IAA on PPRV proliferation and distribution in the host cells were observed by plaque formation experiment, quantitative real-time PCR (qRT-PCR), western-blot (WB) and indirect immunofluorescen assay respectively. The effects of IAA on the intracellular reactive oxygen species (ROS) content, malondialdehyde content and activity of superoxide dismutase (SOD) of the virus-infected cells were respectively evaluated by fluorescent probe, thiobarbituric acid and hydroxylamine colorimetry method, and the effects of IAA on the expression levels of PI3K, AKT, p-AKT, Bcl-2 and Bax in the PPRV-infected cells were detected by Western-blot. [Result] The results showed that the maximum non-toxic mass concentration, the CC50 on Vero cells, the EC50 on PPRV and the TI were 7.81, 125.60, 21.02 mg·L-1 and 5.98, respectively. The cytopathic effect induced by PPRV infection was significantly inhibited by IAA treatment and the viability of the virus infected cells was significantly higher than that of PPRV-infected cells. The number of plaque formation units and the expression levels of viral structural protein F and N were significantly decreased. The virions in cytoplasm of vero cells were also decreased significantly. Moreover, the activity of SOD of the PPRV-infected cells was significantly increased by IAA, and the content of ROS and MDA was accordingly decreased. The expression levels of PI3K and AKT were significantly up-regulated, whereas the experssion of p-AKT was significantly down-regulated and the Bax/Bcl-2 ratio was also significantly decreased in the PPRV-infected cells due to IAA treatment. [Conclusion] IAA exerts its antiviral activity against PPRV by regulating the expression of PI3K/AKT pathway and downstream molecules Bax and Bcl-2, which was closely related to apoptosis of the virus infected cells, and alleviating oxidative stress caused by virus infection.
Authors and Affiliations
Haojie JIAO, Shaohong CHEN, You LIU
Keywords
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- EP ID EP769171
- DOI 10.16445/j.cnki.1000-2340.20220329.001
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