When exposed to chronic hypoxia or toxin monocrotaline (MCT), female animals develop less severe pulmonary arterial hypertension (PH) compared to males; ovariectomy (OVX) exacerbates PH, and OVX animals treated with estradiol (E2) develop less severe disease. There is a line of evidence suggesting that cardiovascular protective effects of E2 are mediated by its major metabolite, 2-methoxyestradiol (2ME). Recently, we have shown that 2ME attenuates the development and retards the progression of MCT-induced pulmonary hypertension in male rats. We hypothesized that the protective effects of E2 in experimental PH are mediated by 2ME. Subsets of intact and OVX female rats were injected saline (Cont and OXV groups) or MCT (60 mg/kg; MCT and OVX-MCT groups) and some of OVX-MCT animals were treated with 2ME (10 microg/kg/h via osmotic minipumps; OVX-MCT+2ME). After 28 days, MCT caused PH, i.e., increased right ventricular peak systolic pressure (RVPSP) and right ventricle/left ventricle+septum (RV/LV+S) ratio, induced inflammatory response in the lungs and caused media hypertrophy (media thickness and % media index) and adventitia widening of small size pulmonary arteries. Ovariectomy exacerbated the disease, i.e., further increased RVPSP, and RV/LV+S ratio, and augmented vascular remodeling and inflammatory response. In diseased OVX rats, treatment with 2ME prevented the worsening of PH and attenuated the inflammatory response and vascular remodeling. No mortality was recorded in the OVX-MCT+2ME group vs. 10% and 36% mortality in the MCT and OVX-MCT group, respectively. This study suggests that 2-methoxyestradiol (a major non-estrogenic metabolite of E2) may mediate the protective effects of estradiol in MCT-induced PH, and warrants further evaluation of 2ME for treatment of PH.