Fig 2: The aging microenvironment suppresses Ppara expression in murine monocytes(A) EGFP+ BM cells (106) were isolated from young mice and transplanted into wild-type (WT) young or old mice. After 7 days, the CD45+ CD11b+ monocytes were sorted by fluorescence-activated cell sorting (FACS).(B) qRT-PCR analysis of Ppara mRNA expression in EGFP+ (donor) or EGFP- (recipient) blood monocytes isolated from young and old mice (n = 5 per group).(C and D) (C) EGFP+ BM cells (106) were isolated from old mice and transplanted into young and old WT mice. (D) qRT-PCR analysis of Ppara mRNA expression in monocytes (n = 5 per group).(E and F) Relative Tnfa and Il6 mRNA expression in young and old recipient mice, determined by qRT-PCR (n = 5 per group). The data represent the means ± SD.(G and H) Blood monocytes isolated from young mice were daily stimulated with either mouse IL-6 (50–500 ng/mL) or TNF-a (50–500 ng/mL) for 7 days; the relative expression of PPAR-a was detected by qRT-PCR (G) and western blot analysis (H); GAPDH was used as a loading control.(I) The LD level in TNF-a-treated monocytes was detected. *p < 0.05, **p < 0.01, ***p < 0.001; t test.

Fig 3: Resveratrol-ßcd switch tumor-associated macrophage M2-type polarization. (A,C) Typical flow cytometry diagram of TAM (CD11b+F4/80+) or M2 macrophages (CD11b+F4/80+ CD206+). The cells were CD45+ gated. (B,D) The proportions of TAM or M2-like macrophages in the treatment and control groups. Error bar = mean ± S.D. ***p < 0.001. ****p < 0.0001. (E) The numbers of M2-like and (F) M1-like (CD206-F4/80+CD11b+) macrophages per tumor gram. **p < 0.01. (G) THP1 as the control group and THP1-derived M1 macrophages treated with resveratrol-ßcd at the indicated concentrations. TNF and (H) IL6 transcript levels and (I) IL-6 levels in the medium 48 h after treatment were measured to identify M1 polarization. Error bar = mean ± S.D. (J) MRC1, (K) IL10, and (L) IL-10 levels 48 h after treatment with the indicated concentration of resveratrol-ßCd. Error bar = mean ± S.D.

Fig 4: mRNA expression of IL-6 (A, B), IFN-ß1 (C, D) and CCL2 (E, F) in BSMCs from severe asthma (A, C, E) and mild COPD (B, D, F) compared to BSMCs from healthy control groups. n = 4 asthma, n = 3 healthy controls, n = 4 COPD, and n = 3 healthy control smokers. qRT-PCR data is representative of two independent experiments. One way ANOVA using Newman-Keuls multiple comparison test were performed to assess statistical significance between groups. Mean ± SE; (ns) p > 0.05, no significant difference, *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 5: ERK1/2-dependent downstream signalling and cellular responses are regulated by CO2 in ECs. a Increased expression of ICAM-1 and phosphorylation of MSK1 at T581 and NFkB p65 at S276 and S536 in response to 100 µM H2O2 were abrogated by treatment with 10% CO2 (11 min, 4 times, 3-h interval). b The expression of HIF-1a and ICAM-1 were induced by 40 ng/ml RBD and could be decreased by 10% CO2 in ECs (4 incubations, 11 min, 3-h interval). c The phosphorylation of NFkB p65 at S276 was increased in response to RBD, TNFa and IFN? combination and could be decreased by 10% CO2 (4 incubations, 11 min, 3-h interval) in ECs. d–e H2O2-induced (d) and RBD-triggered (e) production of IL-6 by ECs was diminished by 10% CO2 (11 min, 3-h interval). The level of IL-6 secreted into the cell culture supernatant was determined by ELISA. Dimeric intracellular IL-6 was visualised by immunoblotting with an anti-IL-6 antibody, and the expression of this form of IL-6 was normalised to the signal obtained with an anti-GAPDH antibody. f CO2 at a concentration of 9–12% inhibited the production of IL-6 which was induced by RBD, TNFa and IFN? combination. IL-6 production was measured by ELISA. The results of three biological replications are presented as the mean ± SD