RGS2 (Regulator of G-protein signaling-2) -deficient mice exhibit severe hypertension, and genetic variations of RGS2 occur in hypertensive patients. RGS2 mRNA up-regulation by angiotensin II (Ang II) in vascular smooth muscle cells (VSMC) is a potentially important negative feedback mechanism in blood pressure homeostasis, but how it occurs is unknown. Here we demonstrate that Group VIA Phospholipase A2 (iPLA2ß) plays a pivotal role in Ang II-induced RGS2 mRNA up-regulation in VSMC by three independent approaches, including pharmacologic inhibition with a bromoenol lactone (BEL) suicide substrate, suppression of iPLA2ß expression with antisense oligonucleotides, and genetic deletion in iPLA2ß-null mice. Selective inhibition of iPLA2ß by each of these approaches abolishes Ang II-induced RGS2 mRNA up-regulation. Further, using adenovirus-mediated gene transfer, we demonstrate that restoration of iPLA2ß-expression in iPLA2ß-null VSMC reconstitutes the ability of Ang II to upregulate RGS2 mRNA expression. In contrast, Ang II-induced Vasodilator-Stimulated Phosphoprotein (VASP) phosphorylation and Ang II receptor expression are unaffected. Moreover, in wild-type but not iPLA2ß-null VSMC, Ang II stimulates iPLA2 enzymatic activity significantly. Both arachidonic acid and lysophosphotidylcholine, products of iPLA2ßaction, induced RGS2 mRNA up-regulation. Inhibition of LOX, particularly 15-LOX, and COX, but not CytP450 inhibited Ang II- or AA-induced RGS2 mRNA expression. Moreover, RGS2 protein expression was also upregulated by Ang II, and this was attenuated by BEL. Disruption of the Ang II/iPLA2ß/RGS2 feedback pathway in iPLA2ß-null cells potentiated Ang II-induced VASP and Akt phosphorylation in a time-dependent manner. Collectively, our results demonstrate that iPLA2ß participates in Ang II-Induced transcriptional up-regulation of RGS2 in VSMC.