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. 2023 May 25;15(10):4429–4443. doi: 10.18632/aging.204745

Figure 2.

Figure 2

Metformin inhibits inflammation and KGN cell pyroptosis by decreasing ROS production and oxidative stress. (A–E) Cells were pretreated with or without 10 μM LPS for 24 h and then incubated with 20 μM metformin for another 12 h. Intracellular levels of ROS, MDA, SOD, GSH, and GSH-px were measured using respective commercial kits; (F–J) Cells were pretreated with 10 μM LPS for 24 h and then incubated with 20 μM metformin and 10 mM NAC for another 12 h; (F) RT-PCR detected the mRNA levels of NLRP3, caspase-1, ASC, and GSDMD; (G) The protein levels of NLRP3, pro-caspase-1, cleaved-caspase-1, ASC and GSDMD-N were detected by Western blot; (H) Levels of LDH in the cell culture medium were examined using an LDH cytotoxicity detection kit; (I) Caspase-1 activity was assessed using a colorimetric caspase-1 activity assay kit; (J) The levels of IL-1β, IL-18, IL-6, and TNF-α in the cell culture supernatant were determined by ELISA. Data are represented as the means ± SD from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns. not significant.