Dioscin Improves Pyroptosis in LPS-Induced Mice Mastitis by Activating AMPK/Nrf2 and Inhibiting the NF-κB Signaling Pathway

Figure 4

Dioscin reduces pyroptosis of mMECs induced by LPS+ATP. mMECs were treated with Ac-DEVD-CHO (20 μM) for 2 h, followed by dioscin (400 ng/ml) for 2 h, LPS (1 μg/ml) stimulation for 6 h, and ATP (5 mM) was added 30 minutes before the end of stimulation. The mRNA levels of (a) caspase-1, (b) caspase-3, and (j) IL-1β were examined by RT-PCR. (c) The protein levels of (d) caspase-1 and (e) caspase-3 were measured by Western blot and quantified by comparison with β-actin. Cells were treated with Ac-DEVD-CHO (20 μM) for 2 h, followed by dioscin (400 ng/ml) for 2 h, and LPS (1 μg/ml) stimulation for 24 h, and ATP (5 mM) was added 1.5 h before the end of stimulation. (f) The cell death was detected by flow cytometry and analysis of pyroptosis cells. (g) The Annexin⁺ PI ⁺ cells in flow cytometry were quantified. (h) The cell LDH release was detected by LDH kit. (i) The production of IL-1β was detected by ELISA. and compared to the LPS+ATP group; ^& and ^&& compared to the dioscin group.

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