Study On The Sedative-Hypnotic Effect Of Mneyanthes Trifoliata L Extract

Objective:The sedative-hypnotic effects of Menyanthes trifoliata L extract were evaluated in terms of both effectiveness and safety,focusing on its effects on sleep architecture,and the mechanisms were initially investigated in terms of gamma aminobutyric acid(GABA)receptors and 5-hydroxytryptamine(5-HT)neurotransmitters.Methods:1.Extraction and compositional analysis of Menyanthes trifoliata L extract The dried stems and leaves of Menyanthes trifoliata L were extracted with 65% ethanol at reflux and dried under vacuum to obtain Menyanthes trifoliata L extract powder as the drug for subsequent experiments.The main components of the extract were identified by liquid chromatography-mass spectrometry(LC-MS).2.Acute toxicity test of Menyanthes trifoliata L extract The safety of the extract was evaluated by acute toxicity test in terms of behavioral status,body weight change,food intake,water intake,organ morphology,organ coefficient and liver and kidney function.3.Study on the sedative-hypnotic effect and mechanism of Menyanthes trifoliata L extract On the one hand,the sedative effect of Menyanthes trifoliata L extract was evaluated by open field test(OFT),the hypnotic effect was evaluated by pentobarbital sodium synergistically induced reversal reflex experiment and animal model of insomnia induced by caffeine or para-chlorophenylalanine(PCPA),and its effect on sleep structure was evaluated by EEG detection technique.The effects on sleep structure were evaluated by EEG.On the other hand,GABA in the prefrontal cortex and 5-hydroxyindole acetic acid(5-HIAA)and 5-HT in the hippocampus were measured by enzyme linked immunosorbent assay(ELISA),and protein Immunoblotting(Western Blot,WB)was used to detect the expression of GABAA receptors in the hypothalamus,and the mechanism of its sedative-hypnotic effect was initially investigated.Results:1.Extraction and compositional analysis of Menyanthes trifoliata L After 1 kg of dried stems and leaves of Menyanthes trifoliata L were extracted,94.8 g of extract was obtained,with a yield of 9.48%,mainly flavonoids and phenolic acid components.2.Acute toxicity test of the extracts of Menyanthes trifoliata L After the administration of 30 g/kg of the extract(equivalent to 316.46 g/kg of raw drug),the mice did not show any death,lethargy,convulsion tremor,etc.The general condition,body weight,water consumption and food intake did not show any significant abnormality within 14 days,and the blood alanine aminotransferase,glutamic aminotransferase,urea nitrogen and creatinine indexes were normal(P>0.05).3.Study on the sedative-hypnotic effect and mechanism of Menyanthes trifoliata L extract Menyanthes trifoliata L extract could reduce the active distance and the number of traversing lattice in OFT(P<0.05),improve the sleep ratio(P<0.001),shorten the sleep latency(P < 0.001)and prolong the sleep time(P< 0.05)of mice in the pentobarbital sodium synergistically induced rumination reflex experiment,and improve the effects of caffeine(P<0.001)or PCPA(P< 0.001)induced sleep in insomniac mice.EEG results showed that Menyanthes trifoliata L extract improved slow-wave sleep(SWS)in mice(P< 0.05)and reduced the degree of sleep fragmentation,i.e.,the number of W→N(WAKE→NREM),N→W(NREM→WAKE),N→R(NREM→REM),and R→W(REM→WAKE)transitions during sleep was reduced(P < 0.05).In addition,sleep lettuce extract increased the content of GABA in the prefrontal cortex(P < 0.05),5-HT and 5-HIAA in the hippocampus(P < 0.05)and increased the expression of GABAA receptors in the hypothalamus of mice(P < 0.05).Conclusion:1.The extraction rate of the ethanolic extract of Menyanthes trifoliata L was 9.48%,and the composition was mainly phenolic acids and flavonoids.2.No acute toxicity was observed at a dose of 30g/kg of Menyanthes trifoliata L extract.3.Menyanthes trifoliata L extract can improve sleep and regulate sleep structure,and its effect is related to the regulation of GABAA receptors and 5-HT neurotransmitters.