2.7 Forced swim test
The FST test was carried out according to previously reported methods
with slight modification(Zhong et al., 2020), and separate groups of
C57BL/6J mice were used for this test. Briefly, 30 min after a single
injection, mice were individually placed in a glass cylinder (height 45
cm, diameter 20 cm) filled with 25°C water to a depth of 15 cm for 6
min. All mice were forced to swim for 6 min, and the duration of
immobility during the last 4 min interval of the test was recorded by an
investigator blind to the study. The water was replaced after each
trial. A mouse was judged to be immobile when it remained floating in
the water without struggling and only making movements necessary to keep
the nose above the water. The observers were unaware of the treatment of
mice.
2.8 Tail suspension test
The FST is a widely used test for assessing potential
antidepressant-like medications. The TST test was measured according to
the methods described previously(Zhong et al., 2020), and separate
groups of C57BL/6J mice were used for this test. Briefly, 30 min after a
single injection, the test C57BL/6J mice were suspended 60 cm above the
floor for 6 min by adhesive tape placed approximately 1 cm from the tip
of the tail. The duration of immobility was scored between the 2nd and
6th minute for 4 min by an investigator blind to the treatment. Mice
were considered immobile only when they hung passively and were
completely motionless, and any mice that did climb their tails were
removed from the experimental analysis. The observers were unaware of
the treatment of the mice. Means and SEM were calculated for each group.