Reactive oxygen species (ROS) involved in Berberine induced AMPK activation are derived from mitochondria.

Abstract

<p><i>A</i>, Effects of xanthine oxidase inhibitors, allopurinol and oxypurinol, on Berberine-enhanced phosphorylation of AMPK-Thr172 and ACC-Ser79 in BAEC. BAEC were pre-treated with mitochondrial tempol (50 µM) for 30 min, and then treated with Berberine (10 µM) for 2 h. The blots are representative of three independent experiments. <i>B</i>, Effects of adenoviral overexpression of p47phox dominant negative mutants on Berberine-enhanced phosphorylation of AMPK-Thr172 and ACC-Ser79 in BAEC. The blots are representative of three independent experiments. <i>C & D</i>, Effects of mitochondrial tempol (*<i>p</i><0.05, control versus Berberine-treated, #<i>p</i><0.05, Berberine-alone versus Berberine plus mitochondrial tempol). <i>E & F</i>, Effects of UCP-2 siRNA transfection on Berberine-enhanced phosphorylation of AMPK-Thr172 and ACC-Ser79 in BAEC. BAEC were transfected with UCP2 siRNA for 24 hr, and then treated with Berberine (10 µM) for 2 hr. n = 3, *<i>p</i><0.05, control versus Berberine-treated, # <i>p</i><0.05, Berberine-alone versus Berberine plus UCP2 siRNA.<i>G.</i> Effects of adenoviral overexpression of UCP-2 on Berberine-enhanced phosphorylation of AMPK-Thr172 and ACC-Ser79 in BAEC. n = 3 *<i>p</i><0.05, control versus UCP-2; #<i>p</i><0.05, control verus Berberine-alone; +<i>p</i><0.05 Berberine-treated versus Berberine plus UCP2.</p