The elevated plus-maze (EPM) test was carried out to evaluate the anxiety-like behavior after the final drug administration. Rats were transported to the well-ventilated testing room and allowed to acclimate for 30 min. The testing room was illuminated with a red lamp (10 and 25 lux) and kept quiet throughout the behavior test. The EPM (TECHMAN Co., Ltd, China) consisted of two opposite open-arms (50×10×1.5 cm), two opposite enclosed-arms (50×10×40 cm) and a center platform (10×10 cm) connecting four arms in the middle. The maze was elevated to a height of 60 cm above the floor and exposed to a fluorescent lamp suspended at height of 100 cm above the apparatus. At the beginning of the EPM test, the rat was placed in the center platform facing one of the open arms, and then allowed to explore freely for 5 min. The number of entries and time spent in each of the four arms was recorded by an automated video-tracking system (EthovisionXT9, NOLDUS). The proportion of entries and time spent in the open-arms relative to the total entries or time spent in all arms (open/open + closed) were calculated as indices of open-arm exploration. The fewer entries and less time spent in the open arms indicated a higher anxiety state.29,32 After each test, the apparatus was thoroughly cleaned with 30% ethanol solution and dried with paper towels to avoid olfactory trace of the previous animal.

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