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. 2022 Jun 21;13:879690. doi: 10.3389/fphar.2022.879690

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Copyright © 2022 Zhang, Chen, Jia, Liu, Wang, Zhang and Zhen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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SHY-01 rapidly activates AMPK in microglia. (A) BV-2 microglial cells were treated with various concentrations of SHY-01 (SHY) or metformin hydrochloride (Met) for 1 h, PBS was used as vehicle (veh) control. Phosphorylation of AMPK (Thr 172) was detected by Immunoblot. ∗p < 0.05, ∗∗p < 0.01 versus the vehicle control, ^## p < 0.01 represent 0.3 mM Met versus 0.3 mM SHY-01. (B) BV-2 microglial cells were treated with 1 mM SHY-01 (SHY) or Met for indicated time points. ∗p < 0.05, ∗∗p < 0.01 versus the vehicle control. In (A,B), phosphorylated (p)-AMPK (Thr 172) protein levels were expressed relative to vehicle (veh) control as mean ± SD (n = 3) and data were analyzed using one-way ANOVA followed by Dunnett post hoc tests. (C) BV-2 microglial cells were pre-incubated with 1 mM SHY-01 (SHY) followed with LPS (200 ng/ml) challenge for 0, 1, 2, or 4 h. Cells were collected for Immunoblot analysis of p-AMPK (Thr 172), total AMPK and GAPDH. Relative p-AMPK protein levels were expressed as mean ± SD (n = 3), data were analyzed using two-way ANOVA followed by Dunnett’s post hoc tests. ∗∗p < 0.01 versus the LPS group.