2. Establishment of the LiCl-pilocarpine-induced post-SE rat model

The LiCl-pilocarpine-induced post-SE rat model was established as described previously²⁴. Briefly, rats received intraperitoneal (IP) injections of LiCl (127 mg/kg, dissolved in water, Sigma, St. Louis, MO, USA). After 24 hours, scopolamine methyl bromide (1 mg/kg, Sigma-Aldrich, USA) was given IP to the rats to reduce peripheral muscarinic effects. Then, 30 minutes later the muscarinic agonist pilocarpine (40 mg/kg, Sigma-Aldrich, USA) was IP injected to induce SE. Seizures started 10 to 30 minutes after the pilocarpine injection. A modified Racine scale was used to evaluate seizure severity²⁵. The criterion of SE in this study was that recurrent seizures greater than or equal to Racine stage 4 lasted for 30min. At 30 minutes after seizure onset, only rats arriving SE were treated with diazepam (10 mg/kg, Tianjin, China) to terminate seizures, otherwise they were excluded from the experiment.

Pilocarpine administration to rats results in SE, and after a latency period the SRS occurs. According to a previous research, the latency period is about 7.2±3.6d after SE ²⁶, so the rats that survived the first week after SE were monitored with a video surveillance system (a CCD camera, JVC, Japan) to observe their SRS in our study. Every 3–4 rats were kept in a transparent cage, marked with picric acid on their ears or legs to make a distinction with each other. The monitoring period lasted 6 weeks (from the onset of the 2^nd week to the end of the 7^th week after SE induction). Qinglan Chen who was blinded to the grouping and drug administration visually inspected the video-recorded data with 6h/d in the daytime for 6 weeks; only SRS reaching a Racine stage 3 to 5 (rearing and/or rearing and falling) were included for further analysis.