Empagliflozin protects diabetic pancreatic tissue from damage by inhibiting the activation of the NLRP3/caspase-1/GSDMD pathway in pancreatic β cells: in vitro and in vivo studies

Figures & data

Figure 1. Cell viability was determined by using the Cell Counting Kit-8 (CCK8) viability assay. (a) The β TC-6 cell viability was approximately 75% when the glucose concentration was 6 g/L, and the inhibitory effect was strongest when the intervention lasted 16 h. (b) The concentration of EMPA was 200 nmol/L, and the β TC-6 cell activity was highest when the continuous intervention for 16 h. ns P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. β TC-6 cell: Mouse insulinoma islet β cells; CCK8: Cell Counting Kit-8; EMPA: Empagliflozin

Figure 2. The expressions of pyroptosis-related proteins (NLRP3, caspase-1 and GSDMD) of β TC-6 cells in different environment were analyzed by western blotting. (a) The images of electrophoretic bands. (b) β TC-6 cells were under three different treatment conditions as described above for 24 h. ns P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3; GSDMD: Gasdermin D.

Figure 3. The results of immunofluorescence of β TC-6 cells treated with different conditions as described above for 24 h. (a) and (b) Immunofluorescence images showing expressions of NLRP3 and caspase-1 in β TC-6 cells under three different treatment conditions. * P < 0.05; ** P < 0.01; *** P < 0.001

Figure 4. Hematoxylin and Eosin staining of histological sections of mice pancreas. (a) The blank control group. (b) The db/db group. (c) The db/db+EMPA group

Figure 5. The protein expressions of NLRP3, caspase-1 and GSDMD were assumed by immunohistochemistry. (a) Immunohistochemical images and scores of NLRP3 of pancreas in mice treated with different methods. (b) Immunohistochemical images and scores of caspase-1 of pancreas in mice treated with different methods. (c) Immunohistochemical images and scores of GSDMD of pancreas in mice treated with different methods. ns P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3; GSDMD: Gasdermin D.

Figure 6. The results of immunofluorescence of in pancreas of mice treated with different methods as described above for six months. The expressions of NLRP3 (a) and caspase-1 (b) under three different treatment methods by immunofluorescence images and scores. ns P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. NLRP3: nucleotide-binding oligomerization domain-like receptor protein 3.