Role of NAD(P)H oxidase in angiotensin II-induced JAK/STAT signaling and cytokine induction

Inflammatory processes involve both synthesis of inflammatory cytokines, su ch as interleukin-6 (IL-6), and the activation of their distinct signaling pathways, eg, the janus kinases (JAKs) and signal transducers and activator s of transcription (STAT), Superoxide (O-2(-)) anions activate this signali ng cascade, and the vasoconstrictor angiotensin II (Ang II) enhances the fo rmation of O-2(-) anions via the NAD(P)H oxidase system in rat aortic smoot h muscle cells. Ang II activates the JAK/STAT cascade via its type 1 (AT(1) receptor and induces synthesis and release of IL-6. Therefore, we investig ated the rule of O-2(-) anions generated by the NAD(P)H oxidase system on t he Ang II activation of the JAK/STAT cascade and its impact on IL-6 synthes is. Ang II stimulation of rat aortic smooth muscle cells induced a rapid in crease in O-2(-) anions determined by laser fluoroscopy, which can be aboli shed by DPI, a flavoprotein inhibitor. Ang Ii-induced phosphorylation of JA K2, STAT1 alpha/beta, STAT3, and IL-6-synthesis can be abolished by DPI, as determined by immunoprecipitations and Northern blot analysis. Electropora tion of neutralizing antisera targeted against p47(phox), a NAD(P)II oxidas e subunit, abolished Ang II-induced JAK/STAT activation and IL-6 synthesis. Inhibition of JAK2 by its inhibitor AG490 (10 mu mol/L) blocked not only J AK2 activation but also IL-6 synthesis. These results suggest that stimulat ion of the JAK/STAT cascade by Ang II requires O-2(-) anions generated by t he NAD(P)H oxidase system, and O-2(-) anion-dependent activation of the JAK /STAT cascade seems to be additionally involved in Ang II-induced IL-6 synt hesis. Thus, Ang II-induced inflammatory effects seem to require O-2(-) ani ons generated by the NAD(P)H oxidase system.