Attenuation of ROS/Chloride Efflux-Mediated NLRP3 Inflammasome Activation Contributes to Alleviation of Diabetic Cardiomyopathy in Rats after Sleeve Gastrectomy

Figure 5

High glucose-induced ROS overload served as a trigger of NLRP3-mediated pyroptosis in H9c2 cells. (a) Protein expression of SOD2 in heart tissues. (b) mRNA expression of Sod2 relative to Tubb3 in hearts. (c) Flow cytometry quantified the 2-NBDG signals in HG-stimulated H9c2 cells. (d) Glucose concentrations in supernatants. (e) Intracellular ROS levels were assessed using DCFH-DA and flow cytometry. (f) The release of LDH percentage from H9c2 cells was reported to the total LDH release. (g) Caspase-1 activity of H9c2 cells. (h) H9c2 cells with membrane disintegrity were stained with red fluorescence by cell probe E42. (i) TUNEL staining of H9c2 cells. (j) Protein levels of NLRP3-related molecules shown by western blot in H9c2 cells treated with HG or HG+NAC. (k) The ratio of caspase-1 p12/procaspase-1 and (l) phospho-NF-κB/total NF-κB is shown. (m) Relative mRNA expression of Nlrp3-related molecules in H9c2 cells, Tubb3 as the internal reference. Data are presented as . CON vs. HG; ^## HG+NAC vs. HG. in 2-NBDG uptake experiment and in each group in other experiments.

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