Fig. 7: Macrophage glycogen metabolism regulates inflammation in vivo.

a–d C57BL/6 J mice were treated with GPI, 6AN or clodronate liposomes, followed by i.p. injection of 20 μg g⁻¹ body weight LPS. Four hours later, serum levels of UDPG were detected by ELISA, n = 5 mice per group (a). Serum levels of TNF and IL-6 (left and middle, n = 5 mice per group) and NO (right, n = 4 mice per group) were measured by ELISA (b). Rarb, Stat1 and Nos2 expression in peritoneal macrophages was determined by real-time PCR, n = 5 mice per group (c). The long-term survival of LPS-induced peritonitis was assessed, n = 10 mice per group, p values are presented relative to LPS group (d). e Schematic illustration showing the cecal ligation and puncture (CLP). f–j C57BL/6J mice were treated with GPI, 6AN or clodronate liposomes, followed by CLP procedure, the long-term survival of CLP-induced sepsis was recorded, n = 10 mice per group, p values are presented relative to CLP group (f). Serum levels of transaminase AST and ALT (g, n = 4 mice per group), BUN (h, left, n = 5 mice per group), CPK (h, right, n = 4 mice per group), TNF and IL-6 (i, n = 6 mice per group) were determined by ELISA. Peritoneal macrophages were isolated and Rarb, Stat1 and Nos2 expression was determined by real-time PCR, n = 5 mice per group (j). k, l Wide type (WT), P2Y₁₄ (+/−) or P2Y₁₄ (−/−) mice were injected intraperitoneally with 20 μg g⁻¹ body weight LPS or were subjected to a sham or CLP procedure (n = 8 mice per group), the long-term survival was recorded, p values are presented relative to P2Y₁₄ (−/−) group. m–o BMDMs from WT or P2Y₁₄ (−/−) mice were stimulated with IFN-γ/LPS for 24 or 36 h, Rarb, Stat1, Nos2, Tnf, Il6, and Il1b expression was determined by real-time PCR, n = 3 mice per group (m), RARβ, STAT1, iNOS, TNF, and IL-6 expression was determined by western blot (n) and ELISA, n = 6 mice per group (o). Unless otherwise specified, n = 3 independent experiments were performed. Data are presented as mean ± SEM. P values were calculated using one-way ANOVA (a–c, g–j, m and o) and two-sided log-rank (Mantel-Cox) test (d, f, k and l). ****p < 0.0001.