The relevance of resveratrol in ameliorating carotid atherosclerosis through glycolysis

Figures

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  Fig. 1. Changes in food intake, body weight, and blood lipid levels in SD rats. a, b Changes in food intake and body weight among the four groups of SD rats. c, d, e, f The levels of TC, TG, LDL-C and HDL-C level; All data are presented as the mean ± SD, one-way ANOVA, n ≥ 5 per group, *P < 0.05, **P < 0.01, ***P < 0.001 vs. sham group; #P < 0.05, ##P < 0.01 vs. injury group; △P < 0.05: injury group vs. L-Res+injury group and H-Res+injury. Res: Resveratrol; L-Res: low-dose Res (10 mg/kg/day); H-Res: high-dose Res (50 mg/kg/day); Model: balloon injury

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  Fig. 2. Res reduced the level of intimal hyperplasia in SD rats. Representative images and statistical results of neointima in arteries stained with hematoxylin and eosin; 5 × magnification: scale bar: 200 µM and 20 × magnification: scale bar: 50 µM. Res: Resveratrol; L-Res: low-dose Res (10 mg/kg/d); H-Res: high-dose Res (50 mg/kg/d); Model: balloon injury. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 5 per group, *P < 0.05, **P < 0.01, ***P < 0.001 vs. sham group; #P < 0.05, ##P < 0.01 vs. model group

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  Fig. 3. Res inhibits the protein expression of PFKFB3, GLUT1 and HK2. The protein expression levels of PFKFB3、GLUT1 and HK2 in the different groups. (mean ± SD, P > 0.05, one-way ANOVA, n ≥ 6 per group. *P < 0.05, **P < 0.01, ***P < 0.001 vs. sham group;#P < 0.05, ##P < 0.01 vs. model group). Res: Resveratrol; L-Res: low-dose Res (10 mg/kg/day); H-Res: high-dose Res (50 mg/kg/day); Model: balloon injury

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  Fig. 4. Res influences the protein expression of Bax, Bcl-2, Caspase-3 and Cle-Caspase-3. a TUNEL staining results for the four groups of SD rats; 20 × magnification: scale bar: 50 µM; (b) The protein expression levels of Bax, Bcl-2, Caspase-3 and Cle-Caspase-3 in the different groups. (mean ± SD, one-way ANOVA, n ≥ 6 per group. *P < 0.05, **P < 0.01, ***P < 0.001 vs. sham group; #P < 0.05, ##P < 0.01 vs. model group). Res: Resveratrol; L-Res : low-dose Res (10/kg/day); H-Res: high-dose Res (50/kg/day); Model: balloon injury

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  Fig. 5. Assessment of HUVEC viability using the CCK-8 assay. a Cell viability was assessed after treatment with different concentrations of ox-LDL alone. b Cell viability was assessed after treatment with 100 μM ox-LDL and different Res concentrations. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 6 per group, *P < 0.05, **P < 0.01, ***P < 0.001

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  Fig. 6. Levels of glucose uptake and lactate production in HUVECs induced by ox-LDL. a Glucose Uptake Levels in ox-LDL-Induced HUVEC after Addition of Res. b Lactate Production Levels in ox-LDL-Induced HUVEC after Addition of Res. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 6 per group, *P < 0.05, **P < 0.01, ***P < 0.001

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  Fig. 7. Res inhibits glycolysis in the ox-LDL-induced HUVECs. a Expression of glycolysis-related proteins detected by WB analysis. b, c, d The effect of Res on the protein levels of GLUT1, HK2, and PFKFB3, respectively. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 6 per group, *P < 0.05, **P < 0.01, ***P < 0.001

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  Fig. 8. Res inhibits apoptosis in HUVECs treated with ox-LDL. a Expression levels of the apoptosis-related proteins Caspase-3 and Cle-Caspase-3. b Apoptotic cells in the different groups were measured using flow cytometry. The right lower quadrant shows early apoptotic cells, the right upper quadrant shows late apoptotic cells, and the left pper quadrant shows necrotic cells. The apoptosis rate was defined as the earlyapoptosis rate plus the late apoptosis rate. c ROS levels were measured using flow cytometry. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 6 per group, *P < 0.05, **P < 0.01, ***P < 0.001

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  Fig. 9. The effect of Res on the PI3K/Akt signaling pathway in ox-LDL-induced HUVECs. a HUVECs were treated with res at concentrations of 0.25, 0.5, and 1 μM for 24 h; The levels of PI3K, AKT, and their phosphorylated forms were detected in protein samples using WB. b Cells were treated with the AKT inhibitor LY294002, and the levels of p-AKT were detected using WB. c Levels of the key enzyme of glycolysis (HK2, GLUT1, and PFKFB) after addition of the inhibitor. All data are presented as the mean ± SD, one-way ANOVA, n ≥ 6 per group, *P < 0.05, **P < 0.01, ***P < 0.001