Downregulated XBP-1 Rescues Cerebral Ischemia/Reperfusion Injury-Induced Pyroptosis via the NLRP3/Caspase-1/GSDMD Axis

Figure 4

Downregulated XBP-1 restored the cell viability and inhibited cytotoxicity and pyroptosis by OGD/R in HT22 cells. The HT22 cells were, respectively, pretreated with XBP-1 siRNA, 24 h before exposing to OGD/R for 24 h and Z-YVAD-FMK (20 μM) and polyphyllin VI (4 μM) 30 min before exposing to OGD/R for 24 h. (a) Cell viability was detected in HT22 cells by using CCK-8 assay. (b) LDH release was detected in HT22 cells by using LDH assay. (c) The quantity of necrotic cells and pyroptotic cells were analyzed by Annexin V and PI staining in each group. (d) The ratio of pyroptosis was represented the percentage of Annv⁺/PI⁺ cell population. (e) Representative western blots and quantification of indicated proteins in the cellular lysate from HT22 cells in each group. Relative expression was normalized to control group. (f–k) The expression levels of XBP-1s, NLRP3, Caspase-1, GSDMD-N, IL-1β, and IL-18 in HT22 cells after OGD/R. Asterisks indicate statistical significance (, , ). All experiments were repeated for three times.

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