Fig 2: 8-OHG induces macrophage migration and activity.a, b Levels of 4-HNE or 8-OHG in the pancreas (a) or serum (b) in Pdx1-Cre;K-RasG12D/+ (KC) or Pdx1-Cre;K-RasG12D/+;Gpx4-/- (KCG) mice with or without liproxstatin-1 (Lip1) or clophosome (Clop) treatment (n = 3 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). c–e Mouse bone marrow-derived macrophages (BMDMs) or human blood monocyte-derived macrophages (HPBMs) were treated with 4-HNE (500 ng/ml) or 8-OHG (500 ng/ml) or guanosine (“G”, 500 ng/ml) for 24 and 48 h. The cell migration (c) and mRNA levels or release of Il6 (d) and Nos2 (e) were assayed (n = 3; two-way ANOVA with Tukey’s multiple comparisons test). f Co-localization of 8-OHG DNA and CGAS in tumors of KC and KCG mice at 3 months of age (n = 5 mice/group; one-tailed t test). g Expression of CGAS and TMEM173 in KC mice at 3–12 months of age (n = 5 mice/group; two-way ANOVA with Tukey’s multiple comparisons test). h–j BMDMs from wild-type (WT), Tmem173-/-, or Tlr9-/- mice were treated with 8-OHG (500 ng/ml) for 24 and 48 h. The cell migration (h) and mRNA levels of Il6 (i) and Nos2 (j) were assayed (n = 3; two-way ANOVA with Tukey’s multiple comparisons test). Data in a–j are presented as mean ± SD. Data are from two or three independent experiments.

Fig 3: iNOS levels in cells of each group. The levels of iNOS in HL-1 cells in blank control group, OxLDL intervention group, OxLDL + low dose astragaloside IV (12.5 µM) treatment group, and OxLDL + high-dose astragaloside IV (50 µM) treatment group (*** indicates P < 0.001; NS: no significance; n = 5).

Fig 4: HSCs regulate the transformation of M1/M2 macrophages. (a) Western blot showing expression of a-SMA in the aLX-2. Activated HSCs (aLX-2) were obtained by treating LX-2 cells with TGF-ß. (b) Lactate level in aLX-2. (c) The level of glycolysis in aLX-2. (d) Detection of the expression levels of M1 macrophage surface markers IL-1ß, IL-6, iNOS, and TNF-a in transformed THP-1 cells cultured with aLX or its supernatant and their supernatants by ELISA. (e) Detection of the expression levels of M2 macrophage surface markers Arg1, CD163, IL-10, and TGFß in co-aLX-2 co-LX-a cells and their supernatants by ELISA. (f) Western blot showing expression of acetylation n co-aLX-2 co-LX-a cells and their supernatants. *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 5: The effect of the FAG on NO concentration (A), iNOS enzyme activity (B), COX-1 enzyme activity (C), COX-2 enzyme activity (D), and PGE2 concentration (E) before and after treatment of LPS-stimulated RAW264.7 cells with the new FAG. Indomethacin was used as a positive control. Bars represent mean ± SD (n = 3), **; p < 0.01, ***; p < 0.001, when compared to untreated LPS-stimulated RAW264.7 cells.