**3.3 VD3 decreases depression-like behavior in the forced swimming test of long-term OVX rats treated with 17β-E2 exposed to CUMS**

CUMS produced a significant increase of the immobility time and decrease of swimming time in the long-term OVX compared to the non-CUMS/CUMS SHAM rats (**Figure 4**, F(1,34) = 52.84, F(1,76) = 68.89, F(1,76) = 26.12, respectively,

#### **Figure 4.**

*VD3 decreased depression-like behavior in the forced swimming test of long-term OVX rats treated with 17β-E2 submitted to CUMS: (a) immobility time, sec, (b) swimming time, and (c) climbing time, sec. \* – 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.*

*Vitamin D Deficiency*

long-term OVX rats with CUMS.

**17β-E2 exposed to CUMS**

rats with CUMS compared to the OVX rats plus CUMS given with solvent. Supplementation with VD3 (5.0 mg/kg) plus 17β-E2 significantly prevented the reduction of the body weight of long-term OVX rats with CUMS (P < 0.001) compared to the OVX plus solvent or 17β-E2/SHAM rats exposed to CUMS (**Figure 2**, P < 0.001). This effect of co-administration of VD3 (5.0 mg/kg) plus 17β-E2 was similar to the effect of the reference drug fluoxetine (10.0 mg/kg) in

**3.2 VD3 increases sucrose preference in the long-term OVX rats treated with** 

Before the CUMS protocol, there was no significant difference among the experimental groups in the SPT (**Figure 3**). Following 28 days of the CUMS trials, the SHAM rats exhibited a decrease in sucrose preference when compared to the control non-CUMS SHAM group (P < 0.05). The sucrose preference in long-term OVX rats was significantly reduced compared to the non-CUMS/ CUMS SHAM rats (**Figure 3**, F(1,34) = 56.14, P < 0.05). Low dose of 17β-E2 increased sucrose preference in long-term OVX rats with CUMS compared to the OVX group with CUMS plus solvent (**Figure 3**, P > 0.05). Treatment with VD3 at dose of 5.0 mg/kg plus 17β-E2, as well as fluoxetine, markedly increased sucrose

*VD3 increases sucrose preference in the long-term OVX rats treated with 17β-E2 submitted to CUMS: (a) Prior to CUMS and (b) After CUMS. \* – 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.*

**160**

**Figure 3.**

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