Menopause is considered to be a natural change of life, which may be accompanied by various health problems such as osteoporosis and fracture. This study is designed to assess the effect of estrogen treatment, folic acid supplementation on bone markers in ovariectomized (ovx) rats. Fifty adult female rats were divided into five equal groups: one served as control, the rats in the second group were subjected to ovariectomy, the third group were exposed to ovariectomy and treated with estrogen, the fourth group were ovariectomized and supplemented with folic acid and the fifth group were consist of ovariectomized rats treated with estrogen and supplemented with folic acid. Ovariectomized rats showed insignificant increase in serum ca+2 and osteocalcin levels with significant decrease in serum vitamin D3 (vit.D3) and significant increase in alkaline phosphates activity and homocysteine levels. Estrogen treatment of ovariectomized rats showed a significant decrease in serum homocysteine (Hcy) levels, alkaline phosphates activity accompanied with insignificant decrease in serum ca+2, vit.D3 and osteocalcin than ovariectomized rats. Supplementation of folic acid to ovariectomized rats induced significant increase in osteocalcin and decreased Hcy levels, insignificant changes in serum ca+2, vit. D3 and alkaline phosphates than ovariectomized control rats, while combined treatment of ovariectomized rats with estrogen and folic acid exhibit better action on bone turnover with marked decrease in Hcy level. So, we can conclude that, supplementation of folate with estrogen treatment has a beneficial effect for menopausal women to avoid osteoporosis and fracture.