To evaluate the efficacy,mechanism and safety of compound essential oil sniffing on refreshing and alerting mice.The effects of compound essential oil on refreshing and alerting the brains of Kunming mice were comprehensively evaluated through behavioral tests of autonomic activity and sleep induced by pentobarbital sodium.The contents of dopamine (DA) and glutamic acid (GLU) neurotransmitters in the blood and brain of mice were detected by enzyme-associated immunoassay (ELISA).The effects of compound essential oil on dopamine receptor D1R protein levels in different brain regions of mice were studied by Western blot,and the mechanism of alerting the brain was clarified.The safety of compound essential oil was investigated by acute toxicology experiment of single high-dose inhalation administration,and the chemical constituents of compound essential oil were analyzed by gas chromatography and mass spectrometry (GC-MS).The results of animal experiments showed that compared with the control group,the mice in the low,medium and high dose groups of compound essential oil could significantly increase the exercise distance every day after three days of sniffing (P<0.01),average velocity (P<0.01),shorten the rest time (P<0.01),significantly increase the autonomic activity of mice;Sniffing essential oils for three consecutive days significantly shortened the total sleep duration (P<0.01),has the effect of refreshing and waking the mind.ELISA results showed that compared with the control group,compound essential oil significantly increased DA and GLU levels in the blood and brain of mice (P<0.05,P<0.05);Western blot results showed that compound essential oil significantly upregulated D1R protein levels in hippocampus,hypothalamus,prefrontal cortex and other brain tissues of mice (P<0.05).In a single high-dose sniffing experiment,compound essential oil had no significant effects on body weight,blood cell-related indexes,liver and kidney function in mice,and HE stained pathological sections showed that compound essential oil had no pathological damage to brain,nose,lung,liver,kidney and other tissues.Results of GC-MS showed the highest content of L-menthol (24.54%),followed by eugenol (16.25%),L-menthol (7.78%),bornol (7.30%),L-caryophyllene (5.29%) and 1,8-eudesin (3.40%).Compound essential oils have the effect of refreshing and waking the brain in mice,and are safe for sniffing.Its mechanism may be related to increased levels of DA and GLU in the blood and brain,and increased regulation of D1R protein levels in different brain regions.