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

Figure 2

Dioscin ameliorates inflammation and death of mMECs induced by LPS+ATP. mMECs were pretreated with dioscin at different concentrations (100 ng/ml, 200 ng/ml, and 400 ng/ml) for 2 h, and LPS (1 μg/ml) was stimulated for 12 h, and then, ATP (5 mM) was added for 30 min before the LPS stimulation was completed. (a–e) The mRNA levels of IL-1β, IL-6, TNF-α, INOS, and COX-2 were examined by RT-PCR. (f–h) The protein levels of INOS and COX-2 were examined by Western blot. Relative protein levels of them were normalized to β-actin. The mMECs were pretreated with dioscin (400 ng/ml) for 2 h, LPS was stimulated for 24 h, and then ATP (5 mM) was added 1.5 h before LPS stimulation was completed. (i) TUNEL staining of treated mMECs. The scale bar represents 200 μm. The green arrow represents the positive TUNEL cells. (j) mMEC survival rate measured by CCK-8 kit. (k) The cell LDH release was detected by LDH kit. ^# and ^## compared to the control group; and compared to the LPS+ATP group.

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