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

Figure 6

Dioscin reduces LPS+ATP-induced pyroptosis of mMECs by activating Nrf2. mMECs were treated with ML385 (5 μM) for 4 hours before adding dioscin (400 ng/ml) for 2 hours, followed by LPS (1 μg/ml) stimulation for 6 hours, and ATP (5 mM) was added 30 minutes before the end of stimulation. (a–j) The protein of nuclear Nrf2, p65, pp65, GSDMD-N, NLRP3, ASC, pro-caspase-1, caspase-1, and HO-1 was detected by Western blot. mMECs were treated with ML385 (5 μM) for 4 h before adding dioscin (400 ng/ml) for 2 h, followed by LPS (1 μg/ml) stimulation for 24 h, and ATP (5 mM) 1.5 h before the end of stimulation. (k, l) The cell pyroptosis was detected by flow cytometry. (m) Different treatment groups of mMECs were detected, and LDH release was detected by LDH kit. (n) The production of IL-1β was detected by ELISA. ^# and ^## compared to the control group; and compared to the LPS+ATP group; ^& and ^&& compared to the dioscin+LPS+ATP group.

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