Figure 6. Activation of NLRP3 with nigericin abolished Metformin-inhibited the release of pro-inflammatory factor in vitro. (A) Cell viability was measured by MTT assay (n = 6 per group). (B) The supernatant was collected and used to determine the LDH activity (n = 6 per group). (C) Statistical results of TUNEL-positive cells per field. (D) Tunel assay (apoptotic cells stained in green fluorescence) was performed to assess examine the apoptosis rate of NRVM cells in all groups (n = 6 per group). Magnification 200x, Scale bar = 100 μm; (E) Western blots were performed to determine p-AMPK, AMPK, NLRP3, cleaved-caspase 1, IL-1β and GAPDH in the total cell lysates. (F–I) Quantitative analysis of p-AMPK, AMPK, NLRP3, cleaved-caspase 1, IL-1β expression and calculate the ratio of p-AMPK/AMPK (n = 4 per group). (J) The levels of IL-1β, IL-18 and TNF-α in the supernatant from NCVMs were detected by ELISA (n = 6 per group). Values are expressed as the mean ± SEM. * P < 0.05 vs. Con. # P < 0.05 vs. H/R. & P < 0.05 vs. MET+veh.