Abstract
Angiotensin II exacerbates oxidative stress in part by increasing superoxide (O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math>) production by many renal tissues. However, whether it does so in proximal tubules and the source of O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> in this segment are unknown. Dietary fructose enhances the stimulatory effect of angiotensin II on proximal tubule Na+ reabsorption, but whether this is true for oxidative stress is unknown. We hypothesized that angiotensin II causes proximal nephron oxidative stress in part by stimulating NADPH oxidase (NOX)4-dependent O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> production and decreasing the amount of the antioxidant glutathione, and this is exacerbated by dietary fructose. We measured basal and angiotensin II-stimulated O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> production with and without inhibitors, NOX1 and NOX4 expression, and total and reduced glutathione (GSH) in proximal tubules from rats drinking either tap water (control) or 20% fructose. Angiotensin II (10 nM) increased O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> production by 113 ± 42 relative light units·mg protein-1·s-1 in controls and 401 ± 74 relative light units·mg protein-1·s-1 with 20% fructose (n = 11 for each group, P < 0.05 vs. control). Apocynin and the Nox1/4 inhibitor GKT136901 prevented angiotensin II-induced increases in both groups. NOX4 expression was not different between groups. NOX1 expression was undetectable. Angiotensin II decreased GSH by 1.8 ± 0.8 nmol/mg protein in controls and by 4.2 ± 0.9 nmol/mg protein with 20% fructose (n = 18 for each group, P < 0.047 vs. control). We conclude that 1) angiotensin II causes oxidative stress in proximal tubules by increasing O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> production by NOX4 and decreasing GSH and 2) dietary fructose enhances the ability of angiotensin II to stimulate O-2<math><mrow><msubsup><mo>O</mo><mn>2</mn><mo>-</mo></msubsup></mrow></math> and diminish GSH, thereby exacerbating oxidative stress in this segment.