| BackgroundLemon Balm(Melissa Officinalis)from Lamiaceae Melissa is a perennial herb,native to the Mediterranean,mainly grown in France.In ancient medical books it has an effection of antidepressants,anti-anxiety,chills,sweating,fever and pain,etc,and often abroad as a mild sedative,antispasmodic,antibacterial agents.But the study on Lemon Balm especially its antidepressant effect is very limited for the research scholars.ObjectiveTo separate,purificate and identificate the active chemical ingredient of lemon balm leaves,To explore the influence of different crude extracts of lemon balm leaves on depression behavioral in mice.Methods1.Soak and extract the plant Lemon Balm which has been dried with methyl alcohol at room temperature.Conduct the extracting solution with vacuum concentration.Then extract it with petroleum ether,ethyl acetate and aqueous acetone,get the extractum with vacuum concentration.Extract using dextran gel column chromatography,preparative thin layer chromatography,silica gel column chromatography and recrystallization were isolated and purified using spectral analysis(13C-NMR,1H-NMR,1H-1H COSY,HSQC,IR,UV,HRESI-MS,EI-MS)and physical and chemical properties of its direct and indirect analysis to confirm the structure of the compound.2 Experimental groups and the establishment of CUMS Depression ModelSixty male Kun-Ming mice were evaluated with the opening test,weed out unqualified mouse.Eight mice were randomly chosen as the normal control group(A),raising environmental standards.CUMS depression model was established by using other mices.Random sampling of depression after modeling 40 mice were model group(B),fluoxetine group(C),lemon balm leaves petroleum ether extract group(D),lemon balmleaves ethyl acetate extract group group(E),lemon balm leaves aqueous acetone extract(F).3.Behavioral assessmentAssessing the changes of behavior of the mices with sucrose consumption test,open field test and forced swimming test before CUMS stimulation,after 21 d CUMS stimulate,after the intervention seven days,after the intervention fourteen days.4.Statistical method.The experimental data were analyzed with SPSS19.0 statistical software package.Multiple groups of measurement data were processed in one-way analysis of variance(one-way ANOVA).When there was homogeneity of variance,LSD method was applied.Independent samples t test and paired t test were applied when comparing between two groups.P <0.05 was considered as statistically significant.Results1.The results of identification of compoundsAs a result thirteen compounds were isolated and identified from the Lemon Balm,as protocatechuyl ldehyde(1),serratagenic acid(2),vanillin(3),2α,3β-dihydroxy-urs-12-en-28-oic acid(4),ursolic acid(5),oleanolic acid(6),daucosterol(7),2α,3β,23,29-tetrahydroxyolean-12-en-28-oic acid-29-O-β-D-glucopyranoside(8),luteolin(9),rosmarinic acid(10),luteolin-7-O-β-D-glucoside(11),β-stitosterol(12),palmitic acid(13).2.The result of the behavioral assessmentBefore CUMS stimulus,each group of mice has no significant difference in the sucrose water consumption,the opening test,the forced swimming test.(P> 0.05).After 21 days CUMS stress;group A compared with group B,group C,group D,group E,group F,has significant difference in sugar water consumption,the opening test,the forced swimming test(P<0.05).Lemon balm extract intervene 7 days,some behaviors of the mice in group C,group D,group F compared with group have improvement,some behaviors ofthe mice in group C recovered rapidly,the result of group B,compared with group E has no significant difference(P> 0.05);Intervention 14 days,compared with the model group,group C and group D could significantly increase the consumption of sucrose and locomotor activity behavior in mice(P <0.01),and significantly reduce mice immobility time in the FST,group C and group D had no significant difference in each test(P> 0.05),group E had no significant effect(P> 0.05)on the results of SPT,OFT and the FST;group F was significantly increased consumption of sugar and reducing the immobility time in mice(P <0.01),but no significant effect on the activities of autonomous behavior.ConclusionThirteen compounds were isolated from the Lemon Balm,compounds 1-8 were isolated from this plant for the first time,compounds 1-3,8 were isolated from this genus for the first time;compared with fluoxetine group and model group,petroleum ether extract of Melissa officinalis has obvious antidepressant effect,acetone water extract of Melissa officinalis also has antidepressant effect,but ethyl acetate extract of Melissa officinalis showed no antidepressant effect. |
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