**3.5 VD3 alters serum estradiol and VD3 levels in long-term OVX rats treated with 17β-E2 exposed to CUMS**

The ELISA assay revealed decreased estradiol and VD3 concentrations in the long-term OVX rats with CUMS compared to the non-CUMS/CUMS SHAM groups (**Figure 6**, F(1,34) = 78.56, F(1,34) = 56.12, F(1,34) = 22.21, respectively, P < 0.05). Low dose of 17β-E2 induced increase of estradiol levels in the serum blood to some extent in the long-term OVX rats with CUMS, however, it was not statistically significant (P > 0.05). The co-administration of VD3 with 17β-E2 significantly increased estradiol and VD3 concentrations in the longterm OVX rats with CUMS compared to the OVX plus solvent or 17β-E2/SHAM with CUMS rats (**Figure 6**, P < 0.05). Fluoxetine did not change the serum

#### **Figure 6.**

*Vitamin D Deficiency*

groups (**Figure 4**, P < 0.05).

**treated with 17β-E2 exposed to CUMS**

P < 0.05). VD3 (5.0 mg/kg), as well as fluoxetine treatment, significantly reduced the immobility time and increased the swimming time in the long-term OVX treated with 17β-E2 compared to the OVX plus solvent or 17β-E2/SHAM with CUMS

**3.4 VD3 changes the behavior in the open field test of long-term OVX rats** 

Following 28 days of CUMS protocol, there were no statistically significant differences for grooming activities between all the experimental groups of animals in the OFT (**Figure 5**, F(1,34) = 0.82, P > 0.05). The long-term OVX rats with CUMS

*VD3 alters the behavior in the open field test of long-term OVX rats treated with 17β-E2 submitted to CUMS. (a) Crossing, (b) rearing, and (c) grooming. \* – P < 0.05 versus the control group, # – P < 0.05 versus to the SHAM group with CUMS, \$ – P < 0.05 versus to the OVX group with CUMS, and \$\$ – P < 0.05 versus to the OVX group with CUMS treated with 17β-E2. The data are presented as mean ± SD; n = 7 in each group.*

**162**

**Figure 5.**

*VD3 alters serum estradiol and VD3 levels in long-term OVX rats treated with 17β-E2 submitted to CUMS. (a) estradiol, pg/ml and (b) 25-OH-VD3, μg/ml. \* – P < 0.05 versus the control group, # – P < 0.05 versus to the SHAM group with CUMS, \$ – P < 0.05 versus to the OVX group with CUMS, and \$\$ – P < 0.05 versus to the OVX group with CUMS treated with 17β-E2. The data are presented as mean ± SD; n = 7 in each group.*

estradiol and VD3 levels in the long-term OVX rats exposed to CUMS (**Figure 6**, P > 0.05).
