ABSTRACT
Objective: To isolate indoleamine 2,3-dioxygenase (IDO) inhibitory constituents from Rabdosia japonica based on bioactivity tracking separation. Methods: The overground part of R. japonica was extracted with boiling water and precipitated by ethanol, the precipitation was collected and lyophilized to obtain XPS, then successively separated by DEAE Sepharose Fast Flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give XPS10-1. A combination of HPGPC, monosaccharide and amino acid composition analysis and IDO inhibitory studies was performed to investigate the structure and bioactivity of XPS10-1. Results: A IDO inhibitory glycoprotein, XPS10-1, was obtained from R. japonica based on activity tracking, its average molecular weight was estimated to 8 852, monosaccharide composition analysis showed the glycosyl part of XPS10-1 was mainly composed of rhamnose and glucose with the ratio of 10.0:2.2, and the protein part was mainly composed of glutamic acid, serine and glycine with mass ratio of 37.3:16.9:45.8. XPS10-1 showed potent IDO inhibitory effect with IC50 of (46.6 ± 3.4) μg/mL, and IC50 of IDO inhibitory effect of XPS10-1 on HeLa cells was (139.0 ± 8.7) μg/mL. Conclusion: In this study, a glycoprotein with IDO inhibitory effect was isolated from R. japonica, which could lay the foundation for the substance basis study of R. japonica.
ABSTRACT
Pedalitin, isolated from the aerial part of Rabdosia japonica (Labiatae), inhibited soybean lipoxygenase-1 (EC 1.13.11.12, Type I) with an IC50 of 152.5 uM. The progress curves for an enzyme reaction, pedalitin inactivate the lipoxygenase-1 in a time dependent, irreversible manner, exhibiting kinetics with a kinact/KI of 59.6 +/- 10 mM-1min-1. In the pseudoperoxidase activity, pedalitin is very slowly oxidized by the soybean lipoxygenase-1 catalyzed decomposition of lipid hydroperoxides.
Pedalitina, aislada de las partes aereas de Rabdosia japonica inhibió a la lipooxigenasa-1 (EC 1.13.11.12 tipo I) con un IC50 de 152.5 uM. La curva de progreso para una acción enzimática, pedalitina inactivó a la lipooxigenasa-1 de una manera dependiente del tiempo, de una manera irreversible, exhibiendo una cinética con una kinact/KI de 59.6 +/- mM-1min-1. En la actividad pseudoperoxidasa, pedalitina es oxidada lentamente por la descomposición de la lípido hidroperóxido de la lipooxigenasa-1 de poroto de soya.
Subject(s)
Flavones/isolation & purification , Flavones/pharmacology , Isodon/chemistry , Lipoxygenase Inhibitors/pharmacology , Soybeans/enzymology , Kinetics , Lipoxygenase Inhibitors/isolation & purification , Time FactorsABSTRACT
A quick HPLC-ESI-MS/MS method was established for simultaneous determination of four major diterpenoids in Rabdosia japonica var.glaucocalyx, including glaucocalyxin A, oridonin, hebeirubesensin and enmenol. Analysis was performed on an Agilent ZORBAX SB-C18(4.6 mm×250 mm, 5 μm ) column eluted in a gradient program with methanol and water. The flow rate was 0.8 mL•min⁻¹. Multiple reaction monitoring (MRM) scanning mode was performed in negative ion switching mode to apply for the quantitative determination. The calibration curves for the above four compounds were linear in corresponding injection amount. The average recoveries of the compounds ranged from 92.40% to 105.9%, with RSDs of 1.7%-6.5%. The method is simple, rapid, accurate with good repeatability, which can provide a reference for overcalling evaluation the quality of R. japonica var.glaucocalyx. The result of cluster analysis- showed that the quality of R. japonica glaucocalyx var. greatly varied between areas and parts.
ABSTRACT
The present study was designed to evaluate the protective effects of ethanol extracts of Rabdosia japonica var. glaucocalyx (Maxim.) Hara (RJ) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice and the possible underlying mechanisms of action. The mice were orally administrated with RJ extract (16, 32 or 64 mg(kg(-1)) daily for consecutive7 days before LPS challenge. The ung specimens and the bronchoalveolar lavage fluid (BALF) were collected for histopathological examinations and biochemical analyses. Pretreatment with RJ significantly enhanced superoxide dismutase (SOD) activity and reduced the wet-to-dry weight (W/D) ratio, the levels of nitric oxide (NO) and protein leakage, and myeloperoxidase (MPO) activity in mice with ALI, in a dose-dependent manner. RJ reduced complement deposition and significantly attenuated LPS-induced ALI by reducing productions of inflammatory mediators, such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). The results demonstrated that RJ may attenuate LPS-induced ALI via reducing the production of pro-inflammatory mediators, and reducing complement deposition and radicals.
Subject(s)
Animals , Male , Mice , Acute Lung Injury , Drug Therapy , Anti-Inflammatory Agents , Pharmacology , Therapeutic Uses , Antioxidants , Metabolism , Pharmacology , Therapeutic Uses , Complement System Proteins , Metabolism , Inflammation Mediators , Metabolism , Isodon , Chemistry , Lipopolysaccharides , Lung , Metabolism , Nitric Oxide , Metabolism , Peroxidase , Metabolism , Phytotherapy , Plant Extracts , Pharmacology , Therapeutic Uses , Superoxide Dismutase , MetabolismABSTRACT
Objective To extract and identify the chemical constituents of Rabdosia japonica. Methods The constituents were isolated and purified by chromatography on silica gel, Sephadex LH-20 and HPLC. The structures of the compounds were determinedbymass spectrometry (MS) and nuclear magnetic resonance (NMR) spectra1 analysis. Results Totally 10 compoundswere isolated and their structures were identified as four triterpenoids: oleanolic acid (1), ursolic acid (2), 2α-hydro xyursolic acid (3), 2α, 23-dihydroxyursolic acid (4) - six flavonoids: quercetin (5), apiginin (6), luteolin (7), rutin, (18) isorhamnetin (9), and tricin (10). Conclusion Isorhamnetin (9) and tricin (10) had been isolated from this plant for the first time.
ABSTRACT
Objective: To study the diterpenes in the roots of Rabdosia japonica var. glaucocalyx. Methods: The constituents of R. japonica var. glaucocalyx were separated and purified with chromatographic methods, and their structures were elucidated by spectroscopic methods. Results: Two compounds were isolated from the roots of R. japonica var. glaucocalyx, and they were identified as glaucocalyxoside (1) and inflexuside A (2). Conclusion: Compound 1 is a new abietane diterpene glycoside named glaucocalyxoside, and compound 2 is isolated from R. japonica var. glaucocalyx for the first time.
ABSTRACT
AIM: To investigate the chemical constituents of Rabdosia japonica var.glaucocalyx,gathed from Liaoning Province,China METHODS: The ethyl acetate-soluble portion was isolated and purified by means of chromatography.Their structures were identified by spectral analysis. RESULTS: Eight compounds were identified as Glaucocalyxin A(Ⅰ),Glaucocalyxin B(Ⅱ),2?-hydroxyursolic acid(Ⅲ),oeanlic acid(Ⅳ),acacetin(Ⅴ),stigmasterol(Ⅵ),apigenin-7-(O-glucoside)(Ⅶ) and stigmasterol-3-O-glucoside(Ⅷ). CONCLUSION: compounds Ⅴ,Ⅵ,Ⅶ and Ⅷ are isolated from the plant for the first time.