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Shikonin Inhibits the Sirt1/FoxO1/Rab7 Pathway, Regulates Autophagy and Oxidative Stress, and Inhibits Rotavirus Replication and Expansion
Vol 37, Issue 3, 2023
Abstract
Objectives: Rotavirus (RV) is the main pathogen of infantile viral gastroenteritis in the world. The purpose of this study was to explore the mechanism of shikonin inhibiting the replication and activity of RV. Methods: Caco-2 cells were treated with shikonin at different concentrations. The inhibitory effect of shikonin on RV proliferation in vitro was estimated by CytoPathic Effect (CPE) inhibition assay and virus production reduction assay. An autophagy flow detection kit was used to detect the autophagy of different subgroups of cells. The levels of oxidative stress indexes such as reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were estimated. The levels of the Sirt1/FoxO1/Rab7 pathway proteins and autophagy proteins Beclin-1 and LC3II/LC3I were estimated by western blot. Caco-2 cells were treated with Nicotinamide (100 µg/mL), and the role of Shikonin in inhibiting RV was confirmed by salvage experiment. Results: When the concentration of Shikonin was less than 100 µM, there was no obvious cytotoxicity. Shikonin therapy induced dose-dependent inhibition of SA11 and WA virus replication in Caco-2 cells. When Caco-2 cells were infected with RV with different concentrations of the multiplicity of infection (MOI), shikonin produced a similar inhibitory effect on RV. The number of autophagy bodies and autophagy proteins Beclin-1 and LC3II/LC3I decreased notably in shikonin-treated cells. Shikonin can alleviate oxidative stress induced by RV and inhibit RV replication through the Sirt1/FoxO1/Rab7 pathway. Conclusions: Shikonin can inhibit the replication and amplification of RV, which may be related to the inhibition of the Sirt1/FoxO1/Rab7 pathway and the regulation of autophagy and oxidative stress.
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Copyright (c) 2023 Pingping Liu, Chunmiao Chen, Guozhe Zhang, Qi Zhang, Hengyue Yang
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Medical Genetics, University of Torino Medical School, Italy

Department of Biomedical, Surgical and Dental Sciences, University of Milan, Italy