Deleterious effects in cardiovascular system (CVS) have been reported in females with estrogen deficiency. However, studies with hormone replacement therapy (HRT) are not concise. Thus, estrogen receptor independent therapies, such as atorvastatin (ATO) and metformin (MET), could be new strategies to substitute HRT. These drugs have been showed pleiotropic effects, but there are limited data comparing ATO and MET with estrogen on the CVS. This study was performed to evaluate the effects of ATO and MET on mesenteric vascular bed (MVB) reactivity from ovariectomized (OVX) female rats. All procedures were approved by Ethics Committee in Animal Experimentation of the Federal University of Espirito Santo (069/2011). All the surgical experiments were carried out under anesthesia (Ketamine 70 mg/kg + xylazine 10 mg/kg, i.p). Female Wistar rats (180-200g) were divided into five groups (n=6): control (SHAM); ovariectomized (OVX), OVX treated with 17βestradiol (EST, 0.5µg/Kg/day), ATO (ATO, 20mg/Kg/day) or MET (MET, 300mg/Kg/day), 21 days after bilateral ovariectomy. On day 35, the MVB was isolated, perfused and constricted with noradrenaline. Dose-response curves of ACh (10-12 to 10-3 M) were obtained in absence or presence of L-NAME (NOS inhibitor) and L-NAME plus Indometacin (COX inhibitor). Inducible NOS (iNOS), endothelial NOS (eNOS) and NADP(H) oxidase type 2 (NOX 2) protein expression were analyzed by western blot (n=6). The data are reported as means ± SEM, compared by ANOVA. OVX showed decreases in vasodilator response to ACh, and all treatments improves this response (Emax – %relaxation SHAM: 80 ± 3; OVX: 54 ± 4*; EST: 82 ± 3; ATO: 88 ± 5; MET: 68 ± 4; *P<0.05 vs SHAM). L-NAME reduce the relaxation to ACh in all groups (Emax – %relaxation SHAM: 32 ± 8; OVX: 24 ± 7; EST: 34 ± 4; ATO: 35 ± 5; MET: 25 ± 4). L-NAME and Indometacin blockade failed to further reduce ACh-induced relaxation (Emax – %relaxation SHAM: 47 ± 10; OVX: 43 ± 7; EST: 40 ± 7; ATO: 36 ± 7; MET: 25 ± 3). OVX showed elevated levels of iNOS and NOX2 protein expression (iNOS: SHAM: 0.84 ± 0.05; OVX: 0.99 ± 0.04* and NOX2: SHAM: 0.76 ± 0.10; OVX: 1.12 ± 0.13*, *p<0.05), and reduced levels of eNOS (SHAM: 1.07 ± 0.10; OVX: 0.71 ± 0.10**, **p<0.01). All Treatments were able to restore the protein expression values (iNOS: EST: 0.81 ± 0.05; ATO: 0.84 ± 0.03; MET: 0.73 ± 0.04; NOX2: EST: 0.80 ± 0.05; ATO: 0.96 ± 0.09; MET: 0.44 ± 0.07; eNOS: EST: 1.13 ± 0.08; ATO: 1.13 ± 0.09; MET: 0.93 ± 0.05). The relaxation dysfunction caused by estrogen deficiency were improved by ATO and MET treatments, through mechanisms related to nitric oxide pathway, by increase eNOS and decrease iNOS levels and by reduction in NOX2/oxidative stress, providing evidence that non-estrogen therapies could be used to improvement the CVS in estrogen deficient state, such as menopause.