Journal: Biomolecules & Therapeutics

Article Title: γ-Oryzanol Ameliorates Endothelial Replicative Senescence via Downregulation of SGLT2 Expression to Attenuate NADPH-Driven Oxidative Stress

doi: 10.4062/biomolther.2025.133

Figure Lengend Snippet: γ-Orz downregulates SGLT2 expression and reduces NADPH oxidase-induced oxidative stress. (A) Representative blots of SGLT2, p22 phox , NOX2, and eNOS in P3 ECs in presence of γ-Orz (3, 10 µM). (B-E) Cumulative data for expression of SGLT2, p22 phox , NOX2, and eNOS respectively. Data are expressed as mean ± SEM (n=3). # p <0.05, ## p <0.01 vs. P1; * p <0.05; ** p <0.01 vs. P3.

Article Snippet: The protein was loaded onto a 10% sodium dodecyl sulfate–polyacrylamide gel and separated by electrophoresis, then transferred to a polyvinylidene difluoride membrane that was blocked with 5% bovine serum albumin (BSA) for 1 h and incubated overnight at 4°C with primary antibodies against β-actin (sc-47778, 1:5000; Santa Cruz Biotechnology); p53 (sc-6243, 1:1000; Santa Cruz Biotechnology); p21 (sc-397, 1:1000; Santa Cruz Biotechnology); p16 (sc-1661, 1:1000; Santa Cruz Biotechnology); p22 phox (sc-271968, 1:1000; Santa Cruz Biotechnology); SGLT2 (AGT-032, 1:1000, Alomone labs); eNOS (cat. no. 610297, 1:1000; BD biosciences); NOX2 (cat. no. 19013-1-AP, 1:1000; Protein tech); diluted in 5% BSA.

Techniques: Expressing

Journal: Biomolecules & Therapeutics

Article Title: γ-Oryzanol Ameliorates Endothelial Replicative Senescence via Downregulation of SGLT2 Expression to Attenuate NADPH-Driven Oxidative Stress

doi: 10.4062/biomolther.2025.133

Figure Lengend Snippet: Graphical abstract illustrating the protective effects of γ-Orz against replicative senescence in PCAECs. γ-Orz suppresses SGLT2 expression, leading to reduced NADPH oxidase-mediated ROS production, restoration of eNOS levels, and inhibition of the p53/p21/p16 signaling pathway. These effects collectively decrease SA-β-gal activity, improve cell proliferation, and delay ECs senescence, thereby contributing to the mitigation of vascular aging. Abbreviations: γ-Orz, γ-oryzanol; PCAECs, porcine coronary artery endothelial cells; SGLT2, sodium-glucose co-transporter 2; NADPH oxidase, nicotinamide adenine dinucleotide phosphate oxidase; ROS, reactive oxygen species; eNOS, endothelial nitric oxide synthase; SA-β-gal, senescence-associated beta-galactosidase; ECs, endothelial cells.

Article Snippet: The protein was loaded onto a 10% sodium dodecyl sulfate–polyacrylamide gel and separated by electrophoresis, then transferred to a polyvinylidene difluoride membrane that was blocked with 5% bovine serum albumin (BSA) for 1 h and incubated overnight at 4°C with primary antibodies against β-actin (sc-47778, 1:5000; Santa Cruz Biotechnology); p53 (sc-6243, 1:1000; Santa Cruz Biotechnology); p21 (sc-397, 1:1000; Santa Cruz Biotechnology); p16 (sc-1661, 1:1000; Santa Cruz Biotechnology); p22 phox (sc-271968, 1:1000; Santa Cruz Biotechnology); SGLT2 (AGT-032, 1:1000, Alomone labs); eNOS (cat. no. 610297, 1:1000; BD biosciences); NOX2 (cat. no. 19013-1-AP, 1:1000; Protein tech); diluted in 5% BSA.

Techniques: Expressing, Inhibition, Activity Assay