Treatment with quercetin inhibits SARS-CoV-2 N protein-induced acute kidney injury by blocking Smad3-dependent G1 cell-cycle arrest
Wenjing Wu, Wenbiao Wang, Liying Liang, Junzhe Chen, Biao Wei, Xiao-Ru Huang, Professor Xiaoqin Wang, Xueqing Yu, Professor Hui-Yao Lan
Molecular Therapy, doi:10.1016/j.ymthe.2022.12.002
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Author contributions W.W. and W.B. performed the experiment, analyzed the data, and wrote the manuscript; L.L., J.C., B.W. and X.R.H. prepared the animal model and analyzed data; X.W. and X.Y. supervised experiments; H.Y.L. designed, supervised the experiments, and revised the manuscript.
Declaration of interests All the authors declared no competing interests.
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J O U R, n a l P r e -p r o o f Smad3-dependent cell cycle arrest at the G1 phase. (A) Co-IP detects that quercetin dosedependently inhibits the interaction between Flag-N and HA-Smad3 in HEK293T cells coexpressing Flag-SARS-CoV-2 N and HA-Smad3; (B) Western blotting detects that like a Smad3 inhibitor SIS3, pre-treatment with quercetin (64μM) for 24hr is able to inhibit SARS-CoV-2 N protein induced Smad3 signaling in HK-2 cells under high AGE(50µg/ml, 30 mins) conditions ; (C) Western blotting reveals that like a Smad3 inhibitor SIS3, pre-treatment with quercetin (64μM) for 24hr is able to inhibit SARS-CoV-2 N protein induced p-Smad3 nuclear translocation in HK-2 cells under high AGE(50µg/ml, 2hr) conditions; (D-E) Western blotting and real-time PCR demonstrate that like a Smad3 inhibitor SIS3, pre-treatment with quercetin (64μM) for 24hr is able to inhibit SARS-CoV-2 N protein induced upregulation of p16 and cyclin D1 downregulation in HK-2 cells under high AGE (50µg/ml) conditions; (F) Flowcytometry analysis reveals that pretreated with quercetin (64μM) for 24hr is able to inhibit SARS-CoV-2 N protein induced cell cycle arrest at the G1 phase while increasing the cell progression into the S-phase in HK-2 cells under high AGE (50µg/ml) conditions. Data are presented as mean ± SEM for at least three independent experiments
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