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

Figure 3

Dioscin reduces LPS+ATP-induced activation of NF-κB signaling pathway and NLRP3 inflammasome in mMECs. The mMECs were pretreated with dioscin (400 ng/ml) for 2 h before stimulated with LPS for 1 h, and ATP (5 mM) was added 30 minutes before the end of stimulation. (a) Nuclear translocation of NF-κB p65 subunit measured by immunofluorescence. The scale bar represents 200 μm. The red arrow represents the nuclear translocation of NF-κB p65. (b) The phosphorylation of p65 was detected using Western blot. The phosphorylation ratio of NF-κB p65 was quantified. mMECs were pretreated with dioscin for 2 h, then stimulated with LPS for 6 hours, and added ATP (5 mM) 30 min before LPS stimulation was completed. (c, d) GSDMD-N, (e) NLRP3, (f) ASC, (g) caspase-1, and (h) caspase-3 protein expressions were measured using Western blot, expressed as densitometry quantitation using β-actin as an internal control. ^# and ^## compared to the control group; and compared to the LPS+ATP group.

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