## 3 Results
### Restraint Stress Has Induced an Anxiety-like Disorder  
The effect of restraint stress on rats' locomotion tested in OFT and EPM was illustrated in Figure 3. Our results showed that restraint stress exerted no significant effects as compared to the control group on general locomotion in OFT and EPM tests (\(p>0.05\)) (Figure 3a-h). The influence of restraint stress on the emotionality, tested in OFT and EPM, was exemplified in Figure 4. Regarding the locomotor activity and anxiety in OFT, the S6  
Figure 2: Repetitive restraint stress imbalanced the steroids levels and increased the activity of the antioxidant enzymes; (**a**) After repeated exposure to restraint stress, CA3 corticosterone (CS) levels were progressively increased (\(p<0.01\)), whereas (**d**) testosterone (TS) concentrations were proportionally decreased after 2 days (\(p<0.01\)) and 6 days (\(p<0.001\)) of restraining; (**b**) SOD was increased after 2 (\(p<0.01\)) and 6 (\(p<0.05\)) days of stress, while (**c**) CAT was increased \(\sim\)5 fold (\(p<0.001\)) only after 6 days of repetitive stress exposure. The bottom and top of the box are the first and third quartiles and the whiskers above and below the box indicate the 95 th and 5 th percentiles. The median is indicated as a horizontal line. The experimental groups were distributed on the X axis. The Y axis was used to expose the measuring unit for the parameters. Statistical significance between S2 and S6 was noted with # (\(p<0.05\); **\(p<0.01\); **\(p<0.001\)), whereas the comparison to Control was annotated with * (* \(p<0.05\); **\(p<0.01\); ***\(p<0.001\)). F values: 2.75 (CS), 2.25 (TS), 9.05 (SOD), 86.24 (CAT). group made fewer entries and spent less time in the center of open field arena as compared to C group (Figure 4b,c) (\(p<0.05\)). Alongside, EPM has been successfully used to assess anxiety-like behavior in basic research, based on the natural tendency of rodents to explore novel environments. The animals typically spend a greater amount of time exploring the periphery of the arena or the closed arms of a maze, usually in contact with the walls, rather than the unprotected center area. Conversely, more time spent, higher travelled distance, and more entries made in the open arms of the EPM test apparatus during a 5 min test session can be considered as anxiolytic-like behavior [57]. In the EPM, the S6 group exhibited diminished number of entries compared to the S2 group (Figure 4e) (\(p<0.05\)), and it spent less time in the open arms of the EPM apparatus as compared to the control (Figure 4f) (\(p<0.05\)). Taken together, 6 days of restraint stress induced anxiety-like behavior based on EPM and OFT measurements. Figure 4: The effects of restraint stress on emotionality in the open field test (OFT) (**a–c**) and in the  
Figure 3: The effects of restraint stress on total (**a,e**) and peripheral (**b,f**) traveled distance and the total (**c,g**) and peripheral (**d,h**) number of entries in the open field test (OFT) and elevated plus maze (EPM) test. The restraint stress exerted no significant effects on general locomotion in OFT and EPM tests (\(p>0.05\)) (**a–h**). Each group consisted of 10 rats. The bottom and top of the box are the first and third quartiles and the whiskers above and below the box indicate the 95 th and 5 th percentiles. The median is indicated as a horizontal line. The experimental groups were distributed on the X axis. The Y axis was used to expose the measuring unit for the test. elevated plus-maze (EPM) (**d-f**). The S2, and more prominently, the S6 rats exhibited a significantly lower number of entries and spent less time in the central part of the open field arena compared to the Control group (**b,c**) (\(p<0.05\)). In the EPM test, the S6 rats made fewer entries in the open arms compared to S2 animals (**e**) (\(p<0.05\)), whereas 6 days of restraint stress significantly decreased the time spent in the open arms compared to the Control group (**f**) (\(p<0.05\)). Each group consisted of 7 rats.