Fig. 2

Irisin inhibited oxidative stress and pyroptosis in HG-treated Min6 cells by downregulating the expression of NLRP3. Min6 cells were incubated with 25 mmol/L glucose for 24 h to construct a T2DM cell model. The medium was changed to that contained irisin (100 ng/mL), and incubation was continued for 24 h. Next, 3 mM ATP was added to the medium and incubated for 45 min to activate the expression of NLRP3, and then, indices reflecting oxidative stress (A: ROS, B: MDA, and C: SOD) were assessed. (D) GSDMD-N immunofluorescence staining was performed to detect cellular pyroptosis; (E) Western immunoblotting assay was performed to evaluate the level of expression of GSDMD-N, NLRP3, ASC, and C-caspase-1 proteins. (F) Protein quantitative analysis was performed. (G) Caspase-1 activity was determined using the Caspase 1 Activity Assay Kit ; ELISA was performed to determine the level of expression of IL-1β (H) and IL-18 (I) to evaluate the inhibitory effect of irisin on HG-induced oxidative stress and pyroptosis in Min6 cells. Scale bar = 130 μm; **p < 0.01 and ***p < 0.001