Résumé
Objective::To evaluate the effects of Valeriana amurensis roots and rhizomes extract and its active constituents on the activities of six major cytochrome P450 (CYP450) enzymes in human liver microsomes. Method::Coumarin, bupropion, tolbutamide, omeprazole, dextromethorphan and testosterone were used as probe substrates for CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4, respectively. Taking their specific metabolites of hydroxylation or demethylation (7-hydroxycoumarin, hydroxybupropion, 4-hydroxytolbutamide, 5-hydroxyomeprazole, dextromethorphan, 6β-hydroxytestosterone) as indicators of enzyme activities. The analytical indexes were used to establish an in vitro model of human liver microsomes of Cocktail probe substrates. This method was applied to evaluate the effects of V. amurensis roots and rhizomes extract and its active constituents on human liver microsomal enzymes. Result::The V. amurensis roots and rhizomes extract had different inhibitory effects on CYP2B6, CYP2C9, CYP2D6 and CYP3A4, their half-inhibitory concentration (IC50) values were 87.49, 1.73, 68.29, 2.80 mg·L-1, respectively. Among the 9 lignans, (-)-massoniresinol-3α-O-β-D-glucopyranoside had a moderate inhibitory effect on CYP2A6 with an IC50 value of 8.51 μmol·L-1, 8, 8′-dihydroxypinoresinol-4, 4′-di-O-β-D-glucopyranoside had a moderate inhibitory effect on CYP2D6 with an IC50 value of 8.73 μmol·L-1, (+)-medioresinol-4, 4′-O-di-β-D-glucopyranoside had a moderate inhibitory effect on CYP2B6 and CYP2C9 with IC50 values of 5.41 μmol·L-1 and 8.20 μmol·L-1. Conclusion::The V. amurensis roots and rhizomes extract and its active constituents have inhibitory effects on liver CYP450 enzymes. Therefore, in the clinical study of new drugs, it is necessary to fully evaluate the risk of drug interactions caused by combination therapy.
Résumé
Valeriana amurensis Smir. ex Kom. widely distributed in the northeast region of China and some region in Russia and Korea, and its underground parts (roots and rhizomes) being used to cure nervous system diseases such as insomnia. The active components including the essential oil and iridoids of underground parts were investigated in different harvest periods in order to evaluate the quality for the roots and rhizomes of V. amurensis. The content of the essential oil was obtained by hydrodistillation and bornyl acetate in the oil was quantitated by GC-EI. The iridoids, valepotriates were determined by potentiometric titration and the main component, valtrate was quantitated by HPLC-UV. The factors of biomass were considered in the determination of collection period. Statistical analysis of results showed that, the highest content of the essential oil per plant was 22.69 µl in withering period and then 21.58 µl in fruit ripening period, while the highest contents of bornyl acetate, valepotriates and valtrate per plant were 2.82 mg, 31.90 mg and 0.98 mg in fruit ripening period separately. Fruit ripening period was decided as the best harvest period for the content of active constituents and output of drug, and it would provide scientific basis for the artificial cultivation of V. amurensis.
Valeriana amurensis Smir. ex Kom. Se distribuye ampliamente en la regioÌn noreste de China y en algunas regiones de Rusia y Corea, y sus partes subterraÌneas (raiÌces y rizomas) se utilizan para curar enfermedades del sistema nervioso como el insomnio. Se investigaron los componentes activos, incluidos el aceite esencial y los iridoides de las partes subterraÌneas de V. amurensis en diferentes periÌodos de cosecha para evaluar la calidad de las raiÌces y rizomas. El contenido del aceite esencial se obtuvo mediante hidrodestilacioÌn y el acetato de bornilo en el aceite se cuantificoÌ por GC-EI. Los iridoides, valepotriatos se determinaron mediante valoracioÌn potenciomeÌtrica y el componente principal, el valtrato se cuantificoÌ por HPLC-UV. Los factores de biomasa fueron considerados en la determinacioÌn del periÌodo de recoleccioÌn. El anaÌlisis estadiÌstico de los resultados mostroÌ que el mayor contenido de aceite esencial por planta fue de 22,69 µl en el periÌodo de marchitacioÌn y luego de 21,58 µl en el periÌodo de maduracioÌn de la fruta, mientras que el mayor contenido de acetato de bornilo, valepotriatos y valtrato por planta fue de 2.82 mg, 31.90 mg y 0,98 mg, respectivamente, en el periÌodo de maduracioÌn de la fruta por separado. Se definioÌ el periÌodo de maduracioÌn de la fruta como el mejor periÌodo de cosecha para el contenido de constituyentes activos y la produccioÌn de droga, lo cual proporcionariÌa una base cientiÌfica para el cultivo artificial de V. amurensis.
Sujets)
Valeriana/composition chimique , Huile essentielle/composition chimique , Racines de plante/composition chimique , Saisons , Camphanes/analyse , Chromatographie en phase liquide à haute performance , Spectrométrie de masse ESI , Rhizome/composition chimique , Iridoïdes/analyseRésumé
In this study, the chemical constituentsfrom Valeriana amurensis AD-effective fraction were investigated based on the effect of Valeriana amurensis on Alzheimer's disease (AD) in previous study. Valeriana amurensis was extracted with 75% ethanol and the obtained extract were extracted and subjected to AB-8 macroporous resin column to obtain the AD-effective fraction of Valeriana amurensis. 9 compounds (1-9) were isolated with silica gel, ODS column chromatography and preparative HPLC. The structures of these compounds were determined as 6-hydroxy-7-(hydroxymethyl)-4-methylenehexahydrocyclopenta[c]-pyran-1(3H)-one (1), suspensolide F (2), loganin(3), α-morroniside(4), β-morronisid (5), partinovalerosidate (6), zansiumloside A (7), (-)-angelicoidenol-2-O-β-D-glucopyranoside (8), citroside A (9). Compounds 6-9 were isolated from the valerian genus for the first time and further investigated the anti-AD effect of compounds 1-9 in vitro found that compound 2 and 6 protected the PC12 cells from injury significantly.
Résumé
Objective: To screen the active constituents from the anti-AD active fraction from the roots and rhizomes of Valeriana amurensis and to elucidate the therapeutic basis for the neuroprotective effect of the roots and rhizomes in V. amurensis. Methods: The aective fraction of the roots and rhizomes in V. amurensis was separated with various chromatographic processes and the structures of the obtained compounds were identified by physicochemical analysis and various spectra data. The neuroprtective compounds on PC12 cells were screened by MTT assay. Results: Eleven compounds were isolated from the roots and rhizomes of V. amurensis, including five bisepoxy lignans of (+)-medioresinol-4,4'-di-O-β-D-glucopyranoside (1), (+)-syringaresinol-4,4'-di-O-β-D-glucopyranoside (2), prinsepiol-4-O-β-D-glucopyranoside (3), (+)-8,8'-dihydroxy-pinoresinol-4,4'-di-O-β-D-glucopyranoside (4), prinsepiol (5), and 6 iridoids of jatamanin A (6), 7-hydroxy-8-(hydroxymethyl)-4-methylenehexahydrocyclopenta[c] pyran-1(3H)-one (7), 4-hydroxymethyl-cyclopenta[c] pyran-7-carboxaldehyde (8), patriscabroside III (9), jatamanin E(10), and patrinoside (11). The Aβ1-42-induced PC12 cells injuries were alleviated by all the bisepoxy lignans with the concentration of 25, 12.5, and 5 μmol/L. Conclusion: Iridoids 6-10 are isolated from the roots and rhizomes of V. amurensis for the first time. Bisepoxy lignans are the therapeutic basis of neuroprotective effect in the roots and rhizomes of V. amurensis.