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. 2023 Aug 19;8(6):e10590. doi: 10.1002/btm2.10590

FIGURE 2.

FIGURE 2

RIPostC triggered NLRP3 inflammasome activation and pyroptosis of microglia 3 days after AIS. (a) Illustration of the clinical experimental timeline. (b) The Pearson linear correlation analysis of the relevance between GSDMD protein expression and NIHSS score (r = 0.83, p < 0.001). Relative protein expression of GSDMD was assessed by ELISA in 21 AIS patients with RIPostC treatment and 21 AIS patients without RIPostC (Student's t‐test, n = 21). (c) Representative Western blot images of NLRP3, ASC, Pro‐Caspase 1, Caspase‐1, and GSDMD‐N in ischemic penumbra, as well as quantitative analysis of these proteins in determined with β‐actin for normalization (one‐way ANOVA, n = 4). (d) ELISA assays for IL‐1β and IL‐18 in the ischemic penumbra zone of brain tissues (n = 5). (e) Double immunostaining of Iba‐1 and GSDMD revealed a good co‐localization of these two makers. Treatment with RIPostC reduced GSDMD positive microglia in ischemic penumbra. bar = 50 μm (one‐way ANOVA, n = 3). Data presented as mean ± SEM. (f) Representative transmission electron microscopy images of microglia in peri‐infarct area. Magnified views of microglial cytomembrane are marked with dashed line boxes. Bar = 2 μm. Red arrowhead: membrane pores. RPC is the abbreviation of RIPostC in the figure. *p < 0.05; **p < 0.01; ***p < 0.001.