Sweroside Effectively Protects Against Ischemia/Reperfusion Injury by Regulating Excitation-contraction Coupling / 中国实验方剂学杂志

ObjectiveTo explore the effect of sweroside on the protection of cardiac systolic/diastolic function during ischemia/reperfusion （I/R） injury. MethodTwenty-four healthy male SD rats were randomly divided into control group， model group， 10 μmol·L-1 sweroside group and 1 μmol·L-1 digoxin group. The I/R injury was modeled by Langendorff and ligation of the left anterior descending coronary artery. The infarct size in each group was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining and hemodynamic parameters such as left ventricular diastolic pressure （LVDP）， left ventricular end-diastolic pressure （LVEDP）， left ventricular end-systolic pressure （LVESP）， maximum rate of rising of left ventricular pressure （+dp/dtmax） and maximum rate of decreasing of left ventricular pressure （-dp/dtmax） of rat isolated heart were detected by Powerlab. In addition， neonatal rat cardiomyocytes （NRCMs） were isolated and randomly divided into control group， model group， 1 μmol·L-1 sweroside group and 10 μmol·L-1 sweroside group. Hypoxia/reoxygenation （H/R） injury model was established. Cardiac systolic function and calcium transients were examined by multi-functional cell imaging analyzer and laser confocal microscope. Furthermore， real-time polymerase chain reaction（Real-time PCR） was used to verify the mRNA expression of excitation-contraction coupling genes such as L-type calcium channel （Cacnb2）， cytochrome c oxidase subunit 6A2 （Cox6a2）， troponin （Tnnc1， Tnni3， Tnnt2）， actin （Actc1）， and myosin （Myh6， Myl2， Myl4） according to the results of previous transcriptome sequencing and literature investigation. Differentially expressed genes were subjected to cluster analysis. ResultCompared with the conditions in the control group， increased cardiac infarction size （P<0.01） and LVEDP （P<0.01） and decreased LVDP （P<0.01） and LVESP （P<0.05） were observed in the model group， with +dp/dtmax of increasing trend while -dp/dtmax decreasing. Moreover， the cell viability， heart rate and contraction amplitude of NRCMs was reduced （P<0.01）， while the contraction duration， time to peak and relaxation time was elevated （P<0.01） in the model group. Interestingly， sweroside could reverse these indicators （P<0.05）. In addition， the expression of Cacnb2， Cox6a2， Tnnc1， Tnni3， Tnnt2， Actc1， and Myh6， Myl2， and Myl4 was down-regulated in the model group （P<0.05， P<0.01）， but sweroside could up-regulate the expression of the above genes （P<0.05）. ConclusionSweroside effectively regulated Ca2+ level in NRCMs， enhanced cardiac systolic function， and protected against H/R injury by regulating excitation-contraction coupling.

Effect of swertiamarin, gentiopicrin and sweroside on cell apoptosis and expression of Bcl-2 in rheumatoid arthritis fibroblast-like synoviocytes / 中国中药杂志

The aim of this paper was to discuss the effect of swertiamarin, gentiopicrin and sweroside on rheumatoid arthritis fibroblast-like synoviocytes(RA-FLSs) and B-cell lymphoma-2(Bcl-2) and their mechanisms. ZINC database and RCSB PDB database were retrieved for 3 D chemical structures of swertiamarin, gentiopicrin and sweroside and 3 D target protein structures. AutoDock Mgltools 1.5.6, AutoDockVina 1.1.2 and pyMOL 2.2.0 were applied for molecular docking to analyze the relationship between Bcl-2(1 GJH) target protein and important ingredients. The cell apoptosis of RA-FLSs was tested by Annexin V-FITC. The Bcl-2 protein expression of RA-FLSs treated with different ingredients was tested by Western blot. The Bcl-2 mRNA expression of RA-FLSs treated with different ingredients was tested by RT-PCR. Swertiamarin, gentiopicrin and sweroside were docked well with Bcl-2(1 GJH). The binding energy of swertiamarin was-6.9 kcal·mol~(-1), the binding energy of gentiopicrin was-6.7 kcal·mol~(-1) and the binding energy of sweroside was-6.4 kcal·mol~(-1). Compared with the blank group, the Bcl-2 protein expression of each group were reduced, while that of the gentiopicrin group was the highest(P<0.01). Compared with the blank group, the Bcl-2 mRNA expression of each groups were reduced. Gentiopicrin can reduce the Bcl-2 protein expression and the Bcl-2 mRNA expression, so as to promote the RA-FLSs apoptosis.

Terpenoids from aerial parts of Gendarussa vulgaris / 中草药

Objective: To isolate and identify the terpenoids from the aerial parts of Gendarussa vulgaris. Methods: The 95% EtOH extract of the aerial parts of G. vulgaris were isolated and purified by silica gel, Sephadex LH-20, reversed-phase ODS, macroporous adsorption resin AB-8 and semi-preparative high performance liquid chromatography. The compound structures were identified by physicochemical properties and spectroscopic data. Results: Ten terpenoids were identified as gvterpennoid A (1), 4,4,14α- trimethylpregn-8-en-3β,20α-diol (2), betulone (3), ergosterol endoperoxide (4), ursolic acid (5), oleanolic acid (6), 3β-hydroxyl- 11α,12α-epoxy olean-28,13β-lactone (7), sweroside (8), loganin (9), and dehydromorroniaglycone (10). Conclusion: Compound 1 is a new triterpene, named gvterpennoid A. Compound 2 is a new natural product, and its 13C- and 1H-NMR chemical shifts were first completely assigned on the basis of 1D and 2D NMR spectroscopic evidence. Compounds 3-5, 7-10 are isolated from Gendarussa genus for the first time.

Comparative Study on Glycosides Content in Liuwei Dihuang Decoction and Cornus Officinalis - Cortex Moutan Couple / 中国中医药信息杂志

Objective To establish an HPLC method for the content determination of morroniside, sweroside, paeoniflorin and loganin of Liuwei Dihuang Decoction and its Cornus Officinalis-Cortex Moutan couple; To discuss the relationship between the whole prescription and the couple of main pharmacodynamic components. Methods The HPLC method was used at Hypersile C18 column (4.6 mm × 250 mm, 5 μm); the mobile phase consisted of methanol-water (24:76); the detective wavelength was 236 nm; the flow rate was 1.0 mL/min; the column temperature was 30 ℃. Results The linear ranges of morroniside, sweroside, paeoniflorin and loganin were among 0.480–7.680 μg (r=0.999 3), 0.103–1.650 μg (r=0.999 5), 0.120–1.920 μg (r=0.999 1) and 0.227–3.630 μg (r=0.999 7), respectively. The average recovery rates and RSD were 102.79%, 102.29%, 100.99%, 102.48%, and 1.73%, 1.48%, 1.32%, 0.75%, respectively. The contents of morroniside, sweroside and paeoniflorin in Liuwei Dihuang Decoction were slightly higher than that in Cornus Officinalis - Cortex Moutan couple, and the contents of loganin were almost the same. Conclusion The method is simple, stable, accurate and reproducible. It can be used for content determinate of glycosides in Liuwei Dihuang Decoction and Cornus Officinalis-Cortex Moutan couple. Cornus Officinalis-Cortex Moutan couple has the glycosides with tonifying kidney effect of Liuwei Dihuang Decoction.

HPLC fingerprint of Zishen Yutai pills and simultaneous determination of 5 index components / 中国中药杂志

To establish the HPLC fingerprint and determine five index components (loganic acid， chlorogenic acid， loganin， sweroside and asperosaponin Ⅵ) of Zishen Yutai pills by high performance liquid chromatography, and provide a scientific basis for its quality control. The fingerprint chromatogram was analysed by the chromatographic fingerprint similarity evaluation system for tradition Chinese medicine (2012), fifteen common peaks were obtained at the wavelength of 254 nm. Different batches of Zishen Yutai pills showed a similarity of above 0.90 in HPLC fingerprint profiles. For the quantitive analysis method, The separation of five components showed good regression (>0.999 2) with linear ranges, and the mean recoveries were in the range of 97.62%-101.9%, with the RSD (=9) less than 3%. The established fingerprint and quantitative analysis methods are highly specific, simple and accurate, which can reflect the quality of Zishen Yutai pills more comprehensively, and can be used for its quality control.

UPLC fingerprint for three species of folk medicine in Swertia L / 中草药

Objective: To accelerate the development of nation medicine, ultra-high performance liquid chromatography (UPLC) combined with content analysis of index compound, cluster analysis (HCA), and principal component analysis (PCA) was used to evaluate and discriminate three kinds of national folk medicinal plants, Swertia davidi, S. punicea, and S. angustifolia. Methods: The chromatograms of 27 samples from three kinds of Swertia L. were collected. The chromatograms were imported in Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2004A to obtain data retention time and peak area of samples. Three sets of samples of similarity were analyzed. The peak area data were imported in SPSS and SIMCA-P+ software for cluster analysis and principal component analysis, dendrogram and principal component scores were obtained, and the clustering effect was observed. Results: The line relationship of this way was good (R2 > 0.9993), with high precision instrument (RSD of 0.19%-2.45%), the method had good reproducibility (RSD of 0-1.77%), recovery was between 95.8% and 102.3% (RSD of 0.45%-2.81%). Three kinds of Swertia L. medicinal plants were different in the contents of index compounds, and the contents of index compounds in the same species were different. Swertiamarin: S. davidi (0.54-10.05 mg/g), S. angustifolia (undetectable), S. punicea (1.76-15.62 mg/g); gentiopicroside: S. davidi (4.27-11.18 mg/g), S. angustifolia (0.26-3.58 mg/g), S. punicea (1.13-16.11 mg/g); sweroside: S. davidi (0.02-0.28 mg/g), S. angustifolia (0.02-0.14 mg/g), S. punicea (0.11-44.52 mg/g); mangiferin: S. davidi (0.14-0.55 mg/g), S. angustifolia (0.03-0.04 mg/g), S. punicea (0.01-13.49 mg/g). The accuracy of dendrogram classification was 85.2%. The contents of index compounds of three false anomalies were less than the remaining nine samples of S. punicea. The effect of discrimination by PCA scores plot of three Swertia L. plants was good, and S. punicea with dispersive distribution indicated that the individuals were significant difference. Conclusion: The method to evaluate and discriminate three Swertia L. medicinal plants is feasible by UPLC combined with analysis of index compound HCA and PCA. In this article, S. punicea is more valuable compared S. davidi and S. angustifolia.

Contents of seven indicative constituents in seven kinds of Gentiana macrophylla by UPLC / 中草药

Objective: An ultra performance liquid chromatographic (UPLC) method was developed for simultaneously determining seven constituents, such as loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and isovitexin, from the medicinal plants of Gentiana macrophylla. Methods: The separation was performed on an Acquity UPLC® BEH C18 column (50 mm ×2.1 mm, 1.7 μm) through a gradient elution of methanol-0.04% aqueous phosphorite at a flow rate of 0.3 mL/min. The detection wavelength was 242 nm, and the column temperature was set at 30℃. Results: For the seven analytes, loganic acid, swertiamarin, 6'-O-β-D-glucosyl gentiopicroside, gentiopicroside, sweroside, isoorientin, and sovitexin, a good linearity (r≥0.9995) was obtained in the range of 2.100-537.100, 1.050-270.000, 0.920-236.000, 11.100-2830.000, 0.750-192.000, 0.167-102.000, and 0.216-52.800 μg, respectively. Their average recoveries (n=6) were 97.83%-100.08%, respevtively, with RSD values less than or equal to 3.76%. Conclusion: The UPLC method is simpler and more effective than HPLC, and can be used for the simultaneous determination of seven indicative constituents in medicinal plants of G. macrophylla.

One new iridoid from aerial parts of Triosteum pinnatifidum / 中草药

Objective: To study the chemical constituents in the aerial parts of Triosteum pinnatifidum. Methods: The compounds were separated and purified by solvent extraction, thin-layer chromatography and silica gel, Sephadex LH-20, and ODS column chromatography. Their structures were elucidated by extensive spectroscopic methods including 1D and 2D NMR experiments (1H- NMR, 13C-NMR, COSY, HSQC, HMBC, and NOESY) along with HR-ESI-MS analyses and comparison with reference substances. Results: Five iridoids were isolated from the butanol extraction fraction of ethanol extract from the aerial parts of T. pinnatifidum, and identified as triohima C 10-O-β-D-glucopyranoside (1), sweroside (2), loganic acid (3), loganin (4), and grandifloroside (5). Conclusion: Compound 1 is a new compound named triopinoside.

Simultaneous Determination of Five Active Chemical Components in Tibetan Medicine Pterocephalus Hookeri Hoeck by UFLC-PDA / 世界科学技术-中医药现代化

This study was aimed to establish an Ultra Fast Liquid Chromatography-Photo Diode Array (UFLC-PDA) method for the simultaneous determination of five chemical components, which included chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, in Pterocephalus hookeri h eck. Agilent Poroshell 120 SB-C18 (100 mm í 4.6 mm, 2.7 μm) was adopted, with acetonitrile-0.2% phosphoric acid solution in gradient elution as the mobile phase at the flow rate of 1.0 mL·min-1. And the injection volume was 0.4 μL. The detection wavelength was set up at 237 nm and 325 nm. And the column temperature was 30℃. The results showed that the calibration curve was linear within the range of 8.72~218.0, 1.52~38.0, 2.44~61.0, 29.36~734.0, 3.00~75.0μg·mL-1 (r > 0.999 6, n=9) for chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, respectively. The average recovery rates were 99.46%, 99.41%, 100.14%, 98.89%, and 99.42%, respectively. The RSD was 0.69%, 0.66%, 0.60%, 1.21%, and 0.64%, respectively (n = 9). It was concluded that this method was simple, accurate and reproducible, which can be used for the simultaneous determination of the content of five chemical components in P. hookeri.

Chemical constituents from roots of Sambucus williamsii (I) / 中草药

Objective: To study the chemical constituents from the roots of Sambucus williamsii. Methods: Silica gel, ODS, and preparative HPLC were used to isolate the compounds. Their chemical structures were elucidated on the basis of spectral data. Results: Eighteen compounds were isolated, and they were identified as 7α-O-ethylmorroniside (1), 7β-O-ethylmorroniside (2), dehydromorroniside (3), loganin (4), 7-dehydrologanin (5), 7-formyloysecologanin (6), sweroside (7), coniferyl alcohol 9-O-β-D-glucopyranoside (8), 3-methoxy-4-(2-glycerol)-phenylpropanol (9), (7R, 8R)-7, 8-dihydro-9'-hydroxyl-3'-methoxyl-8- hydroxymethyl-7-(4-hydroxyl-3-methoxyphenyl)- 1'-benzofuranpropanol-9'-O-β-D-glucopyranoside (10), (7R, 8R)-4, 7, 9, 9'- tetrahydroxy-3-methoxy-8-O-4'- neoligan-3'-O-β-D-glucopyranoside (threo) (11), (7R, 8R)-3-methoxy-8, 4'-oxyneoligna-3', 4, 7, 9, 9'- pentol (threo) (12), 5-(1'-hydroxyethyl)-methyl nicotinate (13), 3-(hydroxyacetyl) indole (14), 4'-hydroxy-N-(4-hydroxy-3- methoxybenzoyl)-3', 5'-dimethoxybenzamide (15), 3-methoxyl-1H-pyrrole (16), (1S, 3S)-1-methyl-1, 2, 3, 4-tetrahydro-β-carboline-3- carboxylic acid (17), and syringic acid-4-O-α-L-rhamnopyranoside (18). Conclusion: Compounds 8-10 and 13-17 are firstly isolated from the plants in Caprofoliaceae, and furthermore, compounds 1-4, 6, 11, and 18 are isolated from the plants of Sambucus L. for the first time.

Two new secoiridoids from Chelonanthus alatus (Aubl) Pulle (Gentianaceae) / Dos nuevos secoiridoides de Chelonanthus alatus (Aubl) Pulle (Gentianaceae)

The species Chelananthus alatus is an herbaceous plant with known ethno botanical and medicinal properties used in control of fever, especially those produced by malaria. From dried leaves (1.11 Kg), the crude alcoholic extract was fractionated by liquid-liquid partition with different polarity solvents. From the sec-butyl alcohol soluble fraction, by successive application of chromatographic methods, four compounds type iridoid were isolated and identified by spectroscopic techniques. Compound 1 is a new secoiridoid which was identified as sweroside 7-isobutyryloxy, and it is reported here for the first time in the Gentianaceae family; the other secoiridoids which were isolated are known as vogeloside (2), dihydro-chelonanthoside (3) and sweroside (4); vogeloside was identified for the first time in this plant (C. alatus). From the isopropyl acetate extract, in conjunction with the sweroside 7- isobutyryloxy (1), chelonanthoside (5) and sweroside (4), were identified, along with the sweroside 7-isovaleryloxy-(6) as a new side chain isomeric ester of dihydrochelonanthoside (3) . This work presents the spectroscopic analysis of the new structures and some bioactivity data.

La especie Chelonanthus alatus (Gentianaceae) es una hierba de aplicaciones ethnobotánicas reconocidas en medicina tradicional, especialmente en el control de la fiebre producida por la malaria. De las hojas secas (1,11 Kg) se realizó el extracto crudo en alcohol etílico, el cual se fraccionó por partición líquido-líquido (L-L) con disolventes de diferente polaridad. De la fracción soluble en alcohol sec-butílico, se aislaron cuatro compuestos tipo seco-iridoide por aplicación sucesiva de diversos métodos cromatográficos los cuales se identificaron por técnicas espectroscópicas. El compuesto 1 es un nuevo secoiridoide identificado como de 7- isobutiriloxi-swerosido, y se reporta por primera vez en la familia Gentianaceae; los otros tres secoiridoides aislados se conocen como vogelósido (2), dihidrochelonanthosido (3) y swerósido (4); el vogelósido se identificó por primera vez en C. alatus. De la fracción soluble en acetato de isopropilo además del 7-isobutiriloxi-swerosido (1) y el swerosido se aislaron e identificaron, el chelonanthosido (5) y el isovaleriloxi-swerosido (6), el cual es un nuevo isómero del dihidrochelonanthosido. En este trabajo se presenta el análisis espectroscópico que llevó a la elucidación estructural de los compuestos novedosos y algunos datos de bioactividad.

Chemical constituents of Swertia macrosperma / 中草药

Objective: To study the chemical constituents from Swertia macrosperma. Methods: HPLC, column chromatography, and recrystallization techniques were used for the separation and purification of the compounds. Their structures were elucidated by physiochemical properties and spectral analyses. The inhibitory activity against tobacco mosaic virus (TMV) was screened by half-leaf blight spot assay. Results: Eight compounds were isolated from S. macrosperma and their structures were identified as 9, 10-dihydroxyl-sweroside (1), 3'-O-(3-hydroxybenzoyl)-swertiamarin (2), mangiferin (3), 1-O-β-glucopyranosyl-2, 6, 8-trihydroxyl-xanthone (4), campestroside (5), bellidifolin (6), 4, 4'-dihydroxy-Z-stilbene (7), and 4, 4'-dihydroxy-E-stilbene (8). Conclusion: Compound 1 is a new compound named macrospermaoside A, and exhibits weak anti-TMV activity. Compounds 2, 7, and 8 are isolated from the plants of this genus for the first time.

Bioactive glycosides from Chinese medicines

Glycosides are the bioactive components of many famous Chinese medicines. Here reported are some bioactive glycosides we discovered from Chinese medicines in recent years. (1) Pheolic glycosides from Chinese medicines: Gastrodia elata, acontium austroynanense and Helicia erratica, three bioactive phenolic glycosides were discovered and two of them have been developed into new drugs. (2) Terpenoidal glycosides: a) Monoterpenoid: the sweroside from Swertia mollensis has been developed intro an anti-hepatitis drug; b) Diterpenoid: Phlomis betonicoides contains sweet glycoides; c) Triterpenoid: many biologically active triterpenoid glycosides were isolated from Panax plants and Siraitia grosvenorii. (3) Steroidal glycosides: a) C21-steroid: Cynanchum otophyllum and C. atratrum contain anti-epilepsy and-tumor glycosides; b) C27-steroid Hemostatic saponins were found in Paris polyphylla.