Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of a fish oil diet on acetylcholine-dependent vasorelaxation in thoracic aortas. Protein expression of soluble epoxide hydrolase (sEH) in rat aortic strips, rat aorta endothelial cells (RAECs), and rat vascular smooth muscle cells. ( A ) Concentration-vasodilatory response curves induced by acetylcholine in aortic strips obtained from Sprague-Dawley (SD) rats, fed a control diet (black circles, black line) or fish oil diet (open circles, blue line) for 3 weeks. Results are expressed as the mean ± standard error of the mean (SEM) of six strips. ( B ) The protein expression of sEH, platelet endothelial cell adhesion molecule-1 (PECAM-1) (endothelial marker) and β-actin in aortic strips from control or fish oil diet-fed rats. n = 3. ( C ) The protein expression of cytochrome P450 2J2 (CYP2J2) and sEH in RAECs and rat vascular smooth muscle cells was examined by western blotting. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control. IB: immunoblotting.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Expressing, Concentration Assay, Control, Marker, Western Blot

Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA) on sEH protein expression and epoxyeicosatrienoic acid/dihydroxyeicosatrienoic acid (EET/DHET) ratio. ( A , B ) The sEH protein expression in RAECs incubated with 100 μM DHA or EPA for 24 and 48 h was evaluated by western blotting and normalized against β-actin. ( C , D ) RAECs were incubated with 50 or 100 μM DHA or EPA for 48 h, and the sEH protein expression was examined. (E, F) RAECs were incubated with 100 µM DHA or EPA for 48 h, and the intracellular EET/DHET ratio was measured. 11, 12-EET, 14, 15-EET, 11, 12-DHET and 14, and 15-DHET were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Each value represents the mean ± SEM. ( A ) n = 25 (bovine serum albumin (BSA)), 3 (100 µM DHA, 24 h), 22 (100 µM DHA, 48 h); ( B ) n = 13 (BSA), 3 (100 µM EPA, 24 h), 10 (100 µM EPA, 48 h); ( C ) n = 32 (BSA), 10 (50 µM DHA, 48 h), 22 (100 µM DHA, 48 h); ( D ) n = 13 (BSA), 3 (50 µM EPA, 48 h), 10 (100 µM EPA, 48 h); (E, F) n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control. BSA: Fatty acid-free bovine serum albumin.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Expressing, Incubation, Western Blot, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Control

Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of DHA and EPA on intracellular 4-hydroxy hexenal (4-HHE) levels. Effect of 4-HHE on sEH expression. ( A , B ) The concentration of intracellular 4-HHE in RAECs treated with 100 µM DHA or EPA was measured by LC-MS/MS analysis. ( C , D ) RAECs were incubated with 15 or 30 µM 4-HHE for 24 and 48 h, and the protein expression of sEH and β-actin was examined by western blotting. (E–G) RAECs were incubated with 100 µM DHA or EPA, or 15 µM 4-HHE for 24 and 48 h. The relative mRNA level of epoxide hydrolase 2 (EPHX2) was quantified using real-time quantitative PCR (RT-qPCR). Each value represents the mean ± SEM. ( A , B ) n = 3; ( C ) n = 19 (control), 3 (15 µM 4-HHE, 24 h), 16 (15 µM 4-HHE, 48 h); ( D ) n = 25 (control), 16 (15 µM 4-HHE, 48 h), 9 (30 µM 4-HHE, 48 h); (E–G) n = 6 (BSA), n = 3 (100 µM DHA or EPA, 15 µM 4-HHE, 24 or 48 h). * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Expressing, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Control

Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of antioxidants on DHA, EPA, or 4-HHE-suppression of sEH protein expression. ( A – C ) RAECs were pretreated with N-acetyl- l -cysteine (NAC; 10 mM) for 1 h, and then with DHA (100 µM), EPA (100 µM), or 4-HHE (15 µM) for 48 h. The protein expression of sEH and β-actin was evaluated by western blotting. Each value represents the mean ± SEM. n = 7. ( D ) The protein expression of p38 kinase (p38), phospho-p38 (p-p38), heme oxygenase 1 (HO-1), nuclear factor erythroid 2-related factor 2 (Nrf2), and β-actin (in cells treated with 15 or 30 µM 4-HHE for 48 h) was determined by western blotting. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control. NS, no significant difference.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Expressing, Western Blot, Control

Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of Nrf2-knockdown on DHA, EPA, or 4-HHE-suppression of sEH protein expression. RAECs were transfected with Nrf2 Small interfering RNA (siRNA) or control siRNA. After 24 h, the cells were incubated with DHA (100 µM), EPA (100 µM), or 4-HHE (15 µM) for another 48 h. Whole cell lysates were subjected to western blot analysis. ( A ) Nrf2 and β-actin expression was determined by western blotting. ( B – D ) sEH and β-actin expression levels were determined by western blotting. Each value represents the mean ± SEM. n = 6-10. * p < 0.05, ** p < 0.01, *** p < 0.001, compared with control. NS, no significant difference.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Knockdown, Expressing, Transfection, Small Interfering RNA, Control, Incubation, Western Blot

Journal: Nutrients

Article Title: N-3 Polyunsaturated Fatty Acids Decrease the Protein Expression of Soluble Epoxide Hydrolase via Oxidative Stress-Induced P38 Kinase in Rat Endothelial Cells

doi: 10.3390/nu9070654

Figure Lengend Snippet: Effect of the p38 kinase inhibitor (SB203580) on DHA, EPA, or 4-HHE-suppression of sEH protein expression. (A) RAECs were incubated with DHA (100 µM), EPA (100 µM), or 4-HHE (15 µM) for 48 h. p-p38 and p38 levels were determined by western blotting. n = 3. (B–D) RAECs were treated with the p38 kinase inhibitor SB203580 (10 µM) for 30 min before control, DHA (100 µM), EPA (100 µM), or 4-HHE (15 µM) incubation. sEH and β-actin protein levels were determined by western blotting. Each value represents the mean ± SEM. n = 6–7. (B–D) One-way ANOVA followed by Tukey’s HSD post hoc analysis. * p < 0.05, ** p < 0.01. NS, no significant difference.

Article Snippet: Rat aorta endothelial cells (RAECs) were purchased from VEC Technologies Inc. (Rensselaer, NY, USA).

Techniques: Expressing, Incubation, Western Blot, Control

Journal: Nutrients

Article Title: The Potential of L-Arginine in Prevention and Treatment of Disturbed Carbohydrate and Lipid Metabolism—A Review

doi: 10.3390/nu14050961

Figure Lengend Snippet: Effects of L-arginine in the treatment of carbohydrate metabolism disorders.

Article Snippet: Zhang et al. (2020) [ ] , aortic endothelial cells of Sprague-Dawley rats , 1 g/kg bw/day (injection to animals) + 5, 25, 50 mM (isolated cells) , + , L-arginine inhibits the expression of miR-221 and increases the expression of eNOS in cells; L-arginine exerts milder effects than simvastatin, but presumably has fewer side effects , , .

Techniques: Control, Activation Assay, Animal Model, Injection, Concentration Assay, Mouse Assay, Clinical Proteomics

Journal: Nutrients

Article Title: The Potential of L-Arginine in Prevention and Treatment of Disturbed Carbohydrate and Lipid Metabolism—A Review

doi: 10.3390/nu14050961

Figure Lengend Snippet: Effect of L-arginine in the treatment of lipid metabolism disorders.

Article Snippet: Zhang et al. (2020) [ ] , aortic endothelial cells of Sprague-Dawley rats , 1 g/kg bw/day (injection to animals) + 5, 25, 50 mM (isolated cells) , + , L-arginine inhibits the expression of miR-221 and increases the expression of eNOS in cells; L-arginine exerts milder effects than simvastatin, but presumably has fewer side effects , , .

Techniques: Control, Injection, Isolation, Expressing, Animal Model, Concentration Assay, Inhibition, Modification