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OBJECTIVE: To investigate the effects of Rhaponticum uniflorum extract (RUE) on angiogenesis and apoptosis of H22 transplanted tumor tissue in mice. METHODS: Mice bearing H22 hepatoma cells were randomly assigned into 5 groups: model, high, medium and low dose RUE, and 5-fluorouracil (5-Fu) group. The intervention was lasted for 10 d. The pathological changes were detected with hematoxylin & eosin (HE) staining, the apoptosis of hepatoma cells were determined by DNA laddering method, the microvessel densities (MVD) were detected using immunohistochemical assay, and the protein expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2) and hypoxia-inducible factor-1α (HIF-1α) were detected by Western blot method. RESULTS: RUE treatment reduced the cell proliferation, aggravated the necrosis of transplanted tumor tissue, reduced DNA fragmentation of H22 hepatoma cells, decreased MVD of tumor tissue, and down-regulated the protein expression of VEGF, VEGFR2 and HIF-1α of the transplanted tumor tissue, as compared with the model group. CONCLUSION: RUE could exhibit anti-angiogenic and pro-apoptotic effects against H22 hepatoma cells in mice, and its anti-angiogenic mechanism is probably related to down-regulation of VEGF, VEGFR2 and HIF-1α proteins.
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Objective: To investigate the chemical constituents from the flowers of Rhaponticum uniflorum. Methods: The chemical constituents were separated and purified by macroporous resin, silica gel, Sephadex LH-20, and MCI column chromatographies. Their structures were determined by physicochemical properties and spectral data. Results: Seventeen compounds were isolated from the ethanol extract in the flowers of R. uniflorum. Among them, eleven flavones were identified as 5,7,4'-trihydroxy-3'-methoxyflavone (1), quercetin-3'-O-methyl ether (2), apigenin (3), kaempferol (4), luteolin (5), quercetin (6), apigenin-7-O-β-D-glycuronate ethyl ester (7), kaempferol-3-O-α-L-rhamnoside (8), quercetin-3-O-α-L-rhamnoside (9), apigenin-7-O-β-D-glucopyranoside (10), and apigenin-6,8-di-C-β-D-glucoside (11); Two liganans were hemislin B glucoside (12) and hemislin B (13); Two phytoecdysones were ecdysterone (14) and turkesterone (15); Others were ursolic acid (16) and 3,5-O-dicaffeoyl quinic acid (17). Conclusion: Compounds 1, 2, 7-10, 12 and 13 are isolated from the plants of Rhaponticum Cass. for the first time, and the compounds 1, 2, and 7-17 are found from the flowers of R. uniflorum for the first time.
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Objective To establish the quality standard for mongolian medicine Rhaponticum uniflorum (L.) DC. with modern analysis methods. Methods Chlorogenic acid and apigenin were identified by thin -layer chromatography (TLC). The chlorogenic acid content was determined by high performance liquid chromatograghy ( HPLC). And HPLC was performed on Kromat Universil C18 ( 4.6 mm × 250 mm, 5 μm) with methanol-water-acetic acid ( v/v/v, 15:85:0.85) as mobile phase, the detection wavelength was 326 nm, the flow rate was 1.0 mL/min, and the column temperature was 20℃. Results The samples and the reference substance shared the same color spots at the same sites. The linear range of chlorogenic acid was 0.044 66-0.446 6μg (r=0.999 8), and the average recovery rate was 99.84%. Conclusion The method is simple, accurate, and with good reproducibility, and can be used for the quality control of Rhaponticum uniflorum ( L. ) DC.
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OBJECTIVE:To establish HPLC fingerprint sectrum of Rhaponticum uniflorum. METHODS:HPLC was performed on the column of Hypersil-ODS with mobile phase of 0.3% phosphoric acid-acetonitrile(gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 30 ℃ and volume injection was 10 μl. The luteolin was refer-ence,17 batches of R. uniflorum from different production places was analyzed and similarity evaluation system for chromatograph-ic fingerprint of TCM (2004 A edition)was adopted for the similarity analysis. RESULTS:There were totally 11 common peaks with similarity degree≥0.900 of 17 batches. According to the verification,the fingerprint spectrum and reference fingerprint spec-trum of R. uniflorum had good consistency. CONCLUSIONS:The established method is specific and stable,and can provide refer-ence for the identification and quality control of R. uniflorum.
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OBJECTIVE: To investigate the effect of Rhaponticum uniflorum water extract (RUWE) on the oxidative stress and DNA damage of liver with an acute liver injury induced by carbon tetrachloride (CCl4) in mice. METHODS: Mice were randomly assigned to the normal control, model, bifendate (BFD), as well as high and low dose groups of RUWE. Animals were treated once daily for 7 d. At the end of the experiment, CCl4 was given intraperitoneally to the mice of groups, then the alanine aminotransferase (ALT), aspartate aminotransferase (AST), lipid hydroperoxide (LOOH), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), Na+-K+-ATPase and Ca2+-Mg2+-ATPase were detected with the colorimetric method, and DNA damage was analyzed using the electrophoretic method. RESULTS: The administration with RUWE reduced the serum activities of ALT and AST, decreased the LOOH and MDA contents of liver, increased the CAT, GSH-Px, total SOD, Mn-SOD activities and GSH level of liver, increased the Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of liver mitochondria, and reduced the DNA damage of hepatocyte in mice with acute liver injury. CONCLUSION: RUWE has protective effect on acute liver injury induced by CCl4 in mice, probably via reduction of the oxidative stress and DNA damage of liver tissue.