Abstract
β-Sitosterol (24-ethyl-5-cholestene-3-ol) is a common phytosterol Chinese medical plants that has been shown to possess antioxidant and anti-inflammatory activity. In this study we investigated the effects of β-sitosterol on influenza virus-induced inflammation and acute lung injury and the molecular mechanisms. We demonstrate that β-sitosterol (150–450 μg/mL) dose-dependently suppresses inflammatory response through NF-κB and p38 mitogen-activated protein kinase (MAPK) signaling in influenza A virus (IAV)-infected cells, which was accompanied by decreased induction of interferons (IFNs) (including Type I and III IFN). Furthermore, we revealed that the anti-inflammatory effect of β-sitosterol resulted from its inhibitory effect on retinoic acid-inducible gene I (RIG-I) signaling, led to decreased STAT1 signaling, thus affecting the transcriptional activity of ISGF3 (interferon-stimulated gene factor 3) complexes and resulting in abrogation of the IAV-induced proinflammatory amplification effect in IFN-sensitized cells. Moreover, β-sitosterol treatment attenuated RIG-I-mediated apoptotic injury of alveolar epithelial cells (AEC) via downregulation of pro-apoptotic factors. In a mouse model of influenza, pre-administration of β-sitosterol (50, 200 mg·kg−1·d−1, i.g., for 2 days) dose-dependently ameliorated IAV-mediated recruitment of pathogenic cytotoxic T cells and immune dysregulation. In addition, pre-administration of β-sitosterol protected mice from lethal IAV infection. Our data suggest that β-sitosterol blocks the immune response mediated by RIG-I signaling and deleterious IFN production, providing a potential benefit for the treatment of influenza.
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Acknowledgements
This study was funded by the National Natural Science Foundation of China (grant no. 81873065), the Secondary Development Projects of Guangdong Famous and Excellent Traditional Chinese Patent Medicines (grant no. 20174005), the Natural Science Foundation of Guangdong Province (grant no. 2018A030310172), and the China Postdoctoral Science Foundation (grant nos. 2017M622652 and 2019M652987).
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ZFY and NSZ conceived the study; BXZ, ZFY, and NSZ designed the study; BXZ and XLL conducted the in vitro experiments; JL and XPP isolated and analyzed the compound β-sitosterol; BXZ, XLL, HMJ, YBH, and PFX performed animal experiments; and BXZ and JL wrote the paper.
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Zhou, Bx., Li, J., Liang, Xl. et al. β-sitosterol ameliorates influenza A virus-induced proinflammatory response and acute lung injury in mice by disrupting the cross-talk between RIG-I and IFN/STAT signaling. Acta Pharmacol Sin 41, 1178–1196 (2020). https://doi.org/10.1038/s41401-020-0403-9
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