The present preclinical study was created to determine the therapeutic effects of vitamin D hormone treatment as an adjunctive therapy alone or in a combination with low dose of 17β-estradiol (17β-E2) on anxiety-like behavior in female rats with long-term absence of estrogen. Accordingly, the aim of the current study was to examine the effects of chronic cholecalciferol administration (1.0, 2.5 or 5.0 mg/kg subcutaneously, SC, once daily, for 14 days) on the anxiety-like state after long-term ovariectomy in female rats. Twelve weeks postovariectomy, cholecalciferol was administered to ovariectomized (OVX) rats and OVX rats treated with 17β-E2 (0.5 µg/rat SC, once daily, for 14 days). Anxiety-like behavior was assessed in the elevated plus maze (EPM) and the light/dark test (LDT), and locomotor and grooming activities were tested in the open field test (OFT). Cholecalciferol at two doses of 1.0 and 2.5 mg/kg alone or in combination with 17β-E2 produced anxiolytic-like effects in OVX rats as evidenced in the EPM and the LDT, as well as increased grooming activity in the OFT. Our results indicate that cholecalciferol, at two doses of 1.0 and 2.5 mg/kg, has a profound anxiolytic-like effects in the experimental rat model of long-term estrogen deficiency.

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The effects of daily immobilization stress applied to female rats on days 15 to 18 of pregnancy on the activity of the enzyme 5 alpha-reductase (isoform I), involved in the synthesis of brain neurosteroids were studied in male offspring. The results demonstrated a decrease in enzyme activity in the cerebral cortex and hypothalamus of male fetuses one day after the last session of stress, while enzyme activity was elevated in the cortex of neonates. Increases in 5 alpha-reductase activity in the cortex, hippocampus, and hypothalamus were also seen in prenatally stressed males on day 5 of life. There were reductions in plasma testosterone and progesterone levels in experimental animals on day 19 of embryonic life and in neonatal rats, the blood progesterone level in prenatally stressed rats remaining decreased at age five days. The possible involvement of neurosteroids in the actions of prenatal stress on sexual differentiation of the brain is discussed.

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