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Before the last administration of treatments, the mice fasted for 12 h. 6 h after the last administration of treatments, brain tissues of the mice were extracted and weighed. Nine times normal saline was added to the brain tissue, which was then prepared into homogenate by high-speed disperser. The homogenate was centrifuged at 3500 r•min − 1 for 10 min, and the supernatant was taken to obtain 10% brain tissue homogenate. The contents of 5-hydroxytryptamine (5-HT), norepinephrine (NE), dopamine (DA), and Gamma-aminobutyric acid (GABA) in the brain tissue homogenate were determined by ELISA. The possible mechanisms of the sedative-hypnotic effect of EI30 were investigated by comparing 5-HT, DA, NE, and GABA content of the brain tissues of mice in each group.
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