Fig. 3

Irisin decreased oxidative stress levels and pyroptosis through activation of the Nrf2-TrX/TXNIP signaling axis

The T2DM cell model was established via HG induction. Then, 100 ng/mL Irisin was added along with the Nrf2 inhibitor ML385, and the level of oxidative stress of cells in each group was evaluated (A: ROS, B: MDA, and C: SOD). (D) Pyroptosis was detected by GSDMD-N staining. (E) WB assays were performed to evaluate the level of expression of Nrf2 in the nucleus, as well as, the level of expression of TXNIP, TrX1, NLRP3, ASC, C-caspase-1, and GSDMD-N. (F) The protein expression levels were calculated as grayscale values. (G) Caspase-1 activity was assessed. ELISA was performed to assess the level of expression of IL-1β and IL-18 proteins (H) for elucidating the mechanism underlying the inhibition of oxidative stress and NLRP3-mediated pyroptosis by irisin. Scale bar = 130 μm; *p < 0.05, **p < 0.01, and ***p < 0.001