Fig 2: Detection of inflammation and oxidative-reduction related factors in H9c2 cells using ELISA method. (A) iNOS, (B) COX-2, (C) CCL9, (D) CXCL1, (E) CXCL9 and (F) CXCL11 in the culture medium of H9c2 cells. Data are presented as the mean ± SD. Each experiment was repeated three times independently. *P<0.05 vs. NC group. #P<0.05 vs. OT group. DHE, dehydrocostus lactone; TXNIP, thioredoxin-interacting protein; NC, negative control; OT, DHE treatment group; OI, DHE treatment combined with TXNIP inhibition group; OH, DHE treatment combined with TXNIP overexpression group; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; CCL9, C-C motif ligand 9; CXCL, chemokine C-X-C motif ligand.

Fig 3: Detection of pro-inflammatory factors in glial cells. (A) Expression of IL-1b mRNA in NC, DT, DO and DI group of glial cells. (B) Expression of IL-6 mRNA in NC, DT, DO and DI group of glial cells. (C) Expression of TNF-A in NC, DT, DO and DI group of glial cells. (D) Expression of PTGS2 in NC, DT, DO and DI group of glial cells. Data was presented as mean ± SD, each experiment was repeated for three times. *P<0.05 vs NC group of cells. #P<0.05 vs signal dezocine treatment group. NC: normal group, DT: dezocine treatment group, DO: dezocine treatment combined with RAPGEF3 overexpression group, DI: dezocine treatment combined with RAPGEF3 inhibition group. SH: sham group, TD: dezocine treatment group, TO: dezocine treatment combined with RAPGEF3 overexpression group, TI: dezocine treatment combined with RAPGEF3 inhibition group.

Fig 4: Expression levels of inflammation and oxidative-reduction related mRNA in heart tissue of mice using reverse transcription-quantitative PCR. (A) iNOS, (B) TNF-α, (C) IL-1, (D) IL-6, (E) IL-10 and (F) COX-2 in mouse heart tissues. Data are presented as the mean ± SD. Each experiment was repeated three times independently. *P<0.05 vs. NC group; #P<0.05 vs. OT group. DHE, dehydrocostus lactone; TXNIP, thioredoxin-interacting protein; NC, negative control; OT, DHE treatment group; OI, DHE treatment combined with TXNIP inhibition group; OH, DHE treatment combined with TXNIP overexpression group; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2.

Fig 5: Effect of PEA and LPS + IFN? treatment of RAW264.7 cells upon mRNA and the protein levels of COX-2. RAW264-7 cells (2.5 × 105 per well) in six-well plates were treated with LPS (0.1 µg/mL) + INF? (100 U/mL) (or vehicle) and either 0 or 10 µmol/L PEA and cultured at 37°C for either 30 min (Panel A), 24 h (Panel B) or 20 h (Panel C); n = 6–16. In Panels A and B, the individual ?Ct values (the difference in threshold cycle between the mRNA of interest and the housekeeper mRNA, in this case RPL19), are shown with the means represented by the bars. The right y-axes show data where the mean value in the absence of either PEA or LPS + IFN? are set to unity. The scale is antilogged since a change of ?Ct of -1 unit corresponds to a doubling of mRNA. Under the figures are the P values (calculated using bootstrapped linear models for the data expressed as ?Ct as described in Methods) for each time point, given the significant interaction LPS x Time in the main analysis (Table 2). ***P < 0.001, NS P > 0.05 for the comparisons shown in Panel A (i.e. where the LPS × PEA interaction was significant). In Panel C, the data are shown on a log scale, since the variances were unequal for the untransformed data. An one-way ANOVA on the log-transformed data gave F2,15 = 163, P < 0.0001. The residual Q.Q plot was consistent with a normal distribution of the residuals. ***P < 0.001, NS P > 0.05 for the comparisons shown (Holm–Sidak's multiple comparison test).