PMC

Effects of culture medium on iNOS expression and NO production in the course of macrophage activation in F-12 or DMEM. (A) Western blot analysis of iNOS protein and β-actin using total protein extracts of JA-4 cells, cultured in F-12 or DMEM, in the presence or absence of IFN-γ (10 units/mL) and/or LPS (100 ng/mL). (B) Relative amount of iNOS protein. The results in (A) are quantitatively shown as values relative to those for the untreated cells in F-12 after normalization as to the amount of β-actin in each sample. Densitometry data are expressed as means±S.D. for three independent experiments. *p<0.05 between indicated pairs of F-12 and DMEM. (C) Western blot analysis of iNOS protein in total protein extracts of JA-4 cells treated with LPS+IFN-γ in either F-12 or DMEM at 37 °C for the times indicated on the abscissa. (D) Time-course of iNOS protein induction in F-12 and DMEM. The results in (C) are quantitatively shown as values relative to those for the untreated cells in F-12 at 0 h after normalization as to the amount of β-actin in each sample. Densitometry data are expressed as means±S.D. for three independent experiments. *p<0.05 between indicated pairs of F-12 and DMEM. (E) in the culture supernatants of macrophages after incubation with LPS+IFN-γ for the indicated times. was estimated by the Griess reagent assay. The results are shown as means±S.D. for three independent experiments. (F) Expression of iNOS mRNA during macrophage activation in F-12 and DMEM. mRNA was extracted and quantitated by qRT-PCR, as described in the text. The results are relative to those for the untreated cells in F-12 at 0 h after normalization as to the internal control, GAPDH mRNA. The results are expressed as means±S.D. for three independent experiments. *p<0.05 between indicated pairs of F-12 and DMEM.

Relief of depressive-like behaviors and induction of hyperalgesia by intra-VTA infusion of morphine or baclofen in CMS mice. (A1) In the OFT, morphine increased line crossings in CMS mice at 4h and 1d (stress × time epoch interaction: F_{(12, 112)} = 2.99; Bonferroni post-hoc test displays increased line crossings in CMS-Mor(VTA) group relative to the CMS-Sal(VTA) group, *p < 0.05, n = 8 mice/group). (A2) In the TST, morphine decreased immobility duration in stressed mice at 4h and 1d (stress × time epoch interaction: F_{(12, 112)} = 6.5; Bonferroni post-hoc test shows decreased immobility duration in CMS-Mor(VTA) group relative to the CMS-Sal(VTA) group, **p < 0.01, ***p < 0.001; n = 8 mice/group). (A3) In the social interaction test, morphine prolonged the interaction of CMS mice at 4h (stress × time epoch interaction: F_{(12, 112)} = 2.58; Bonferroni post-hoc test demonstrates increased interactive behaviors in CMS-Mor(VTA) group relative to the CMS-Sal(VTA) group, ***p< 0.001, n = 8 mice/group). (A4) In the sucrose preference test, morphine increased the 1% sucrose preference in CMS mice at 4h and 1d (stress × time epoch interaction: F_{(12, 112)} = 3.25; Bonferroni post-hoc test displays increased sucrose consumption in CMS-Mor(VTA) group relative to the CMS-Sal(VTA) group, *p < 0.05, **p < 0.01, n = 8 mice/group). (B) Intra-VTA infusion of morphine reduced the PWL in CMS mice at 0.5 and 1h (stress × time epoch interaction: F_{(12, 112)} = 4.18; Bonferroni post-hoc test indicates hyperalgesia in CMS-Mor(VTA) group relative to the CMS-Sal(VTA) group, ***p < 0.001, n = 8 mice/group). (C) The experimental groups for studying the effects of intra-VTA injection of morphine on depressive-like behaviors and nociceptive phenotype. (D) Experimental timeline. (E1) In the OFT, baclofen increased line crossings in CMS mice at 4h (stress × time epoch interaction: F_{(12, 112)} = 10.12; Bonferroni post-hoc test reveals more locomotion of CMS-Bac(VTA) group than the CMS-Sal(VTA) group, *p < 0.05, n = 8 mice/group). (E2) In the TST, baclofen decreased the immobility duration in CMS mice at 4h (stress × time interaction: F_{(12, 112)} = 5.99; Bonferroni post-hoc test: **p < 0.01 vs. CMS-Sal(VTA) group, n = 8 mice/group). (E3) In the social interaction test, baclofen increased the interaction of CMS mice at 4h (stress × time interaction: F_{(12, 112)} = 3.9; Bonferroni post-hoc test: *p < 0.05 vs. CMS-Sal(VTA) group, n = 8 mice/group). (E4) In the sucrose preference test, baclofen increased the 1% sucrose preference in CMS mice at 4h (stress × time interaction: F_{(12, 112)} = 8.28; Bonferroni post-hoc test: **p < 0.01 vs. CMS-Sal(VTA) group, n = 8 mice/group). (F) Intra-VTA infusion of baclofen reduced the PWL in stressed mice at 0.5 and 1h (stress × time interaction: F_{(12, 112)} = 2.38; Bonferroni post-hoc test: **p < 0.01 vs. CMS-Sal(VTA) group, n = 8 mice/group). (G) The experimental groups for exploring the effects of intra-VTA injection of baclofen on depressive-like behaviors and nociception. (H) Experimental timeline, including CMS paradigm, stereotaxic surgery and behavioral tests.

Plasma LDL-C (A), HDL-C (B), ApoA-I (C), A-II (D), B (E), C-II (F), C-III (G), and E (H) concentrations following ingestion of 40 g of fat with either placebo, 100 mg and 150 mg PP, or 100 mg PP + CC in healthy adults (n = 50). Data are presented as means ± SD and compared with a condition (4 levels) × time (5 levels) repeated-measures linear mixed model, with significance accepted at P ≤ 0.05. The condition–time interactions were as follows—LDL-C: F (12, 924.981) = 0.353, P = 0.979; HDL-C: F (12, 910.981) = 0.349, P = 0.979; ApoA-I: F (12, 926.255) = 0.385, P = 0.969; ApoA-II: F (12, 897.232) = 1.102, P = 0.355; ApoB: F (12, 890.554) = 0.301, P = 0.989; ApoC-II: F (12, 924.441) = 0.628, P = 0.820; ApoC-III: F (12, 928.687) = 0.317, P = 0.987; and ApoE: F (12, 918.277) = 0.503, P = 0.503. CC, caffeine and catechins; PP, polymerized polyphenol.

HL-1 cells were cultured at least during 3 weeks with normal glucose (5.5 mmol/l) then submitted to LG, NG, HG or IHG for 12h with medium change every 2h or 72h with medium change every 12h (see ). The MMP was measured using TMRE in the absence (basal, A) or in the presence of either succinate (B), palmitate (C) or pyruvate (D). Results were normalized to the NG condition. Data are the means ± S.E.M of 3–7 independent experiments. Two-way ANOVA followed by the Tukey’s multiple comparisons test when evaluating the effect of glucose and time treatment *, P < 0.05, vs LG; ^$, P < 0.05, ^$ $ $, P < 0.001, vs NG; ^§, P < 0.05, ^§§§, P < 0.001, 12 vs 72h. For basal, F _{12 vs 72h} (1, 20) = 41.98, P<0.0001; F_{glucose treatment} (3, 20) = 3.239, P = 0.0438; F _{12 vs 72hXglucose treatment} (3, 20) = 10.56, P = 0.0002; For succinate, F _{12 vs 72h} (1, 32) = 3.722, P = 0.0626; F_{glucose treatment} (3, 32) = 7.572, P = 0.0006; F _{12 vs 72hXglucose treatment} (3, 32) = 0.7248, P = 0.5447; For palmitate, F _{12 vs 72h} (1, 36) = 2.257, P = 0.1417; F_{glucose treatment} (3, 36) = 1.543, P = 0.2200; F _{12 vs 72hXglucose treatment} (3, 36) = 2.886, P = 0.0489; For pyruvate, F _{12 vs 72h} (1, 28) = 0.4816, P = 0.4934; F_{glucose treatment} (3, 28) = 2.624, P = 0.0701; F _{12 vs 72hXglucose treatment} (3, 28) = 3.349, P = 0.0331.