FIGURE 4.

CD73 attenuates microglia pyroptosis via PI3K/AKT/Foxo1 signal. After overexpression plasmid transfection for 24 hours, BV2 cells were challenged with LPS (1μg/mL) for another 8 hours, then 3 independent samples, and their controls were RNA sequenced. A, The heat map analysis of differentially expressed mRNAs. B, The KEGG ontology enrichment analyses of differentially expressed mRNAs. C, The mRNA expression of PI3K, AKT, Foxo1 in BV2 cells challenged with/without LPS in presence or absence of pc‐CD73 or MK2206 (3μM) (N = 3). D and E, The expression and phosphorylation of PI3K, AKT, mTOR, Foxo1, IKK‐β, GSK‐3β, Bad and statistical comparison in BV2 cells in different treated groups (N = 3). F and G, The expression and phosphorylation of PI3K, AKT, Foxo1 and statistical comparison in BV2 cells in different treated groups (N = 3). H, The release of LDH was detected by LDH Assay Kit in different cultured groups (N = 5). I, The microglial pyroptosis markers (NLRP3, ASC, CASP‐1, and GSDMD) mRNA expression was identified by RT‐PCR in CD73 upregulated BV2 cells in presence of MK2206 (3μM) or not (N = 3). J, The release of IL‐1β, IL‐18, and TNF‐α in different cultured groups measured by ELISA (N = 3). K and L, The pyroptosis‐related protein expression and statistical comparison in BV2 cells in different treated groups (N = 3). ^* P < .05, ^** P < .01, ^*** P < .001 versus control; ^# P < .05, ^## P < .01, ^### P < .001 versus pc‐CD73. Intraperitoneal injection of 40 mg/kg SC79 or DMSO was carried out every day after surgery in CD73 KO mice. M, Representative immunohistochemical staining for GSDMD and CASP‐1 on the third day after SCI or sham surgery. N, Degree of motor disturbance assessed by the Basso Mouse Scale (BMS) criteria at different time points after SCI with SC79 treatment or not (N = 6). ^* P < .05, ^** P < .01. All data are shown as the mean ± SD independent experiments