FIGURE 6.

Activating SIRT1 with resveratrol reduces NOX2‐dependent oxidative stress and relieves progerin‐associated premature senescence. Freshly primary HSECs, isolated from normal rats and cultured in vitro, were treated with H₂O₂ (10 μM) and were simultaneously administered with resveratrol (a specific chemical activator of SIRT1, 1 μM) for two days. (A) Representative immunoblots of SIRT1, vWF and NOX2 of primary HSECs on Day 2 in four groups (CTR, H₂O₂, H₂O₂ + Resveratrol, Resveratrol). The relative protein expression was quantified in the graph, right. ^* P < .05 vs the CTR group; ^# P < .05 vs the H₂O₂ group. (B) The H₂O₂ content of primary HSECs on Day 2 in four groups (CTR, H₂O₂, H₂O₂ + Resveratrol, Resveratrol). ^* P < .05 vs the CTR group; ^# P < .05 vs the H₂O₂ group. (C) Fluorescence intensity for mito‐SOX of primary HSECs on Day 2 in four groups (CTR, H₂O₂, H₂O₂ + Resveratrol, Resveratrol), measuring with flow cytometry. ^* P < .05 vs the CTR group; ^# P < .05 vs the H₂O₂ group. (D) The SA‐β‐Gal activity in HSECs on Day 2 in the four groups (CTR, H₂O₂, H₂O₂ + Resveratrol, Resveratrol) was observed by SA‐β‐Gal staining (Scale bar: 25 μm). The black triangles indicated the SA‐β‐Gal‐positive cells. The SA‐β‐Gal‐positive cells were quantified in the graph, right. ^* P < .05 vs the CTR group; ^# P < .05 vs the H₂O₂ group. (E) Representative immunoblots of Ac p53 K381, total p53, progerin, Lamin A/C and Lamin B1 of primary HSECs on Day 2 in four groups (CTR, H₂O₂, H₂O₂ + Resveratrol, Resveratrol). The ratio of Ac p53 K381 and total p53 protein levels, as well as the relative protein expression of progerin, Lamin A/C and Lamin B1 were quantified in the graph, right. ^* P < .05 vs the CTR group; ^# P < .05 vs the H₂O₂ group