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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2016 Oct 27;36(12):2394–2403. doi: 10.1161/ATVBAHA.116.308321

Figure 4. High glucose and palmitate downregulate Sirt1 in endothelial cells by inducing miR-34a.

Figure 4

A, MiR-34a in HUVECs is upregulated by high glucose (HG, 30 mM glucose). HUVECs were incubated in HG for 24 h. MiR-34a expression is expressed relative to cells incubated in mannitol (30 mM) control. B, Inhibition of miR-34a rescues HG-induced downregulation of Sirt1. HUVECs transfected with miR-34a inhibitor (miR-34a-I) or scrambled miR were treated with mannitol or HG for 24 h. Data are expressed relative to cells transfected with scrambled miR and treated with mannitol control. C, HG incubation upregulates miR-34a in miR-34afl/fl, but not e-miR-34a−/−, aortic endothelial cells. Aortic endothelial cells isolated from miR-34afl/fl and e-miR-34a−/− mice were treated with mannitol or HG for 24 h. Data is expressed relative to miR-34afl/fl cells treated with mannitol. D, MiR-34a knockout inhibits HG-induced downregulation of Sirt1. Aortic endothelial cells were incubated in HG for 24 h as above. E, MiR-34a regulates basal Sirt1 expression in endothelial cells. HUVECs were transfected with miR-34a mimic (miR-34a-M), miR-34-I, or scrambled miR. Representative immunoblots are shown. F, Palmitate upregulates miR-34a in miR-34afl/fl aortic endothelial cells. Aortic endothelial cells were incubated in 500 uM palmitate for 12 h. MiR-34a expression is shown relative to miR-34afl/fl endothelial cells incubated in BSA vehicle. G, MiR-34a knockout inhibits palmitate-induced downregulation of Sirt1. MiR-34afl/fl and e-miR-34a−/− aortic endothelial cells were treated with 500 uM palmitate for 12 h. Data are expressed relative to miR-34afl/fl endothelial cells treated with BSA vehicle. Representative Immunoblots are shown. Data are shown as mean ± SEM. * P<0.05. n = 3–7.