2.3 Experimental design
The present experiment was divided into two parts (Fig. 1B). In part 1,
effects of FFA on CA/CPR-treated WT mice were investigated. At 1 h after
ROSC, mice were randomized to receive FFA or vehicle by using random
number table (Fig. 1A). Mice in the FFA group were given FFA (12.5 mg/kg
in sterile saline, 4% PEG-400, 2% DMSO; Sigma, St. Louis, MO) via
intraperitoneal injection once daily for 1 week [24], while mice in
the vehicle group received equivalent volume of vehicle solution. Forty
mice with successful resuscitation were followed up for 7 days to
evaluate survival, neurological function, and histological injuries. The
other 45 mice were used to describe the neuroprotective effects by
detecting cellular apoptosis, neuronal degeneration, BBB disruption,
edema formation and neuroinflammation markers at day 3 after CA/CPR. In
part 2, we explored whether gene deletion of Trpm4 improved
neurologic outcome after CA/CPR and whether FFA interfered with the
TRPM4 channel. Twenty-six Trpm4−/− mice that
underwent CA/CPR were randomly allocated to receive vehicle or FFA, and
13 CA/CPR-treated WT mice given vehicle were set as the control group
(Fig. 1A). In this part, expression of TRPM4 was evaluated at 24 h after
CA and other experiments were in line with that of part 1.