| Nardosachys jatamansi DC.is a kind of perennial herb of the genus Nardosachys in Valerianaceae.Its dry roots and rhizomes are commonly used in traditional Chinese medicine,with a long history of medication.It has the functions of regulating qi,relieving pain,relieving stagnation-syndrome and invigorating spleen.Modern pharmacological studies have found that Nardosachys jatamansi DC.possesses significant antidepressant activity,and our previous studies found that many constituents in Nardosachys jatamansi DC.can regulate the activity of serotonin transporter(SERT).Since SERT is an important target for prevention and treatment of digestive tract and neuropsychiatric diseases.Therefore,based on previous studies on the fingerprint-activity relationship of Nardosachys jatamansi DC.regulating the SERT activity,the distribution characteristics of SERT-active constituents of Nardosachys jatamansi DC.were clarified by fitting the curve of "component-activity relationship" of Nardosachys jatamansi DC.Then,this fitting curve allowed us the method optimization of component preparation,and the further isolation and identification of the constituents in the active components..The constituents were screened for SERT activity,and the selectivity of these strongly active constituents targeting SERT was also investigated.Finally,the effects of active component 1 and the total extract on animal behavior were investigated by acute stress model of mice,and their antidepressant effects was evaluated.1.Fitting curve of " component-activity " relationship of Nardosachys jatamansi DC.On the basis of previous studies on the relationship between constituent fingerprint and SERT activity regulated by Nardosachys jatamansi DC.,20 components(Fr.1 to Fr.20)of Nardosachys jatamansi DC.were further prepared by HPLC,and the effect of each component on SERT activity was investigated.With the SERT screening activity data of each component,and the middle value of the retention time of each component in the UPLC-PDA spectrum,the relationship curve between component chemical fingerprint and SERT activity was fitted as a scatter plot.While the pull-in full-span UPLC chromatogram of the total extract gave us an intuitive "component-activity" relationship of Nardosachys jatamansi DC.Assisted by this "component-activity" relationship,the preparation method was optimized and components 1 and 2 were then afforded.Considering that component 1 showed potent SERT activity of 1.21 ± 0.04 ***(mean ± SEM),SERT enhancers must be rich in this component.2.Isolation and structure identification of chemical constituents assisted by the "fingerprint-activity" relationshipSince the total extract and the active component 1 of Nardosachys jatamansi DC.significantly enhanced SERT activity,herein,further isolation and structure identification of the consituents were conducted for characterizing the chemical constituents in the total extract and active component 1.the derived components of Fr.10,Fr.11,Fr.13,Fr.14,Fr.16,and Fr.17 of the total extract and the optimized component 1 were further isolated and purified to obtain sixteen compounds,and by comparisons of their physico-chemical properties and NMR data with those reported data of the known standards prepared in our lab,their structures were identified respectively as 7-oxonardinoperoxide,desoxo narchinol A,kansone B,nardosinonediol,kansone A,1-hydroxy-aristolone,debilon,nardosinone,kansone H,1,8,9,10-tetradehydroaristolan-2-one,1(10)-aristololene-2-one,chlorogenic acid,8?-dihydrogeniposide,7-dioxy-8-epi-loganic acid,adoxosidic acid,8-epi-loganic acid.SERT screening tests revealed that eleven of the sixteen compounds could significantly enhance SERT activity.In order to investigate the selectivity of desoxo narchinol A and chlorogenic acid(the final concentrations were 0.1 ?M,1.0 ?M and 10.0 ?M,respectively)on SERT,they were co-incubated with with the SERT inhibitor,fluoxetine with the final concentration of 2.0 n M.The SERT screening results were desoxo narchinol A with values of 1.12 ± 0.03 ***,1.11 ± 0.02 ** and 1.12 ± 0.03 ***,and chlorogenic acid with values of 1.15 ± 0.03 ***,1.29 ± 0.03 *** and 1.19 ± 0.04 *** respectively.While the SERT activity values of these two compounds were then to be 1.07 ± 0.03,1.09 ± 0.02 and 1.09 ± 0.04* for desoxo narchinol A,and 1.01 ± 0.04,1.09 ± 0.04 and 1.05 ± 0.03 for chlorogenic acid,respectively,when co-incubated with 2.0 n M of fluoxetine.The results revealed that desoxo narchinol A and chlorogenic acid could antagonize the SERT inhibitory activity of fluoxetine and showed a selectivity to SERT target.3.Antidepresant effects of the total extract and the active component 1 of Nardosachys jatamansi DC.in vivoBoth the total extract and component 1 showed strong SERT enhancing activities.Therefore,the acute stress model of mice(open field test and tail suspension test)was used to investigate the antidepressant effects of the total extract and the active component 1 in vivo.Animals were randomly divided into the following groups: the low,medium and high dose groups of the total extract,the blank control group(distilled water: 30 mg/kg/10 m L)and the positive drug control group(imipramine hydrochloride: 30 mg/kg/10 m L);and the low,medium and high dose groups of component 1,the blank control group and the positive drug control group.The results of OFT(open field test)of the total extract were 40.62 ± 5.47,41.66 ± 3.62,47.58 ± 3.45,53.52 ± 2.99,36.48 ± 4.88 *,while the TST(tail suspension test)results of the total extract were 121.10 ± 16.06 *,127.40 ± 11.70 *,113.70 ± 12.26 **,187.40 ± 16.14,76.72 ± 18.99 ***.For the active component 1,the OFT results were 52.58 ± 3.78,49.29 ± 4.56,53.98 ± 6.90,54.83 ± 3.40,35.20 ± 4.99 *,while the TST results were 149.00 ± 20.23,109.90 ± 11.05 **,120.30 ± 12.67 *,187.30 ± 22.67,75.01 ± 14.76 ***.These data indicated that the total extract and component 1 had no significant effects on the spontaneous activity of mice,both of them showed significant antidepressant effects in vivo. |
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