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Objective To observe the effects of amarogentinon liver cancer stem cells (LCSCs) after insufficient thermal ablation and its mechanism. Methods A insufficient thermal ablation model of HepG2 cells was established by water bath method.The percentage of CD133-positive LCSCs and the mRNA and protein levels of CD133 were detected by flow cytometry, qRT-PCR and Western blot.The insufficient thermal ablation model of HepG2 cells was treated with variable doses of amarogentin for 24 h; the percentage of CD133-positive LCSCs, the proliferation and apoptosis of liver cancer cells, and the mRNA and protein levels of CD133, TBC1D15, and p53were detected by flow cytometry, qRT-PCR and Western blot. Results The percentage of CD133-positive HepG2 cells and the mRNA and protein levels of CD133 and TBC1D15in the insufficient thermal ablation model were significantly higher than those in the normal HepG2 cells.Amarogentin then markedly decreased the percentage of CD133-positive LCSCs, the proliferation rate of HepG2 cells, and the mRNA and protein levels of CD133 and TBC1D15 in the insufficient thermal ablationresidual model (all P < 0.05);inversely, the apoptosis rate of HepG2 cells and the phosphorylated levels of p53 in the insufficient thermal ablation model were significantly increased (all P < 0.05). Conclusion Amarogentin could reduce the proportion of LCSCs after insufficient thermal ablation, inhibit the proliferation, and promote the apoptosis of LCSCs, which maybe associated with increasing the phosphorylation of p53 and inhibiting the expression of TBC1D15.
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Objective To study the chemical constituents from the roots of Veratrilla baillonii. Methods Compounds were purified by normal and reversed column chromatographic techniques, and isolated by high performance liquid chromatography. Their structures were identified on the basis of spectral data including MS and NMR. Results Twenty-four compounds (including 10 xanthone glycosides, 8 xanthones, and 6 iridoids) were isolated from the ethyl acetate extracts of the roots of V. baillonii. They were identified as 1-hydroxy-2,3,4-trimethoxyxanthone-7-O-β-D-glucopyranoside (1), tripteroside (2), 1-hydroxy-2,7-dimethoxyxanthone-3-O-β-D- glucopyranoside (3), 1-hydroxy-3,4-dimethoxyxanthone-7-O-β-D-glucopyranoside (4), secamonoide B (5), tetrasweroside A (6), veratriloside B (7), 2,3,4,5-tetramethoxyxanthone-1-O-β-D-glucopyranoxyl-(1→6)-β-D-glucopyranoside (8), 2,3,4,7-tetra- methoxyxanthone-1-O-β-D-xylopyranoxyl-(1→6)-β-D-glucopyranoside (9), 2,3,5-trimethoxy-xanthone-1-O-β-D-xylopyranoxyl- (1→6)-β-D-glucopyranoside (10), 1,3-dihydroxy-4,7-dimethoxyxanthone (11), 1,7-dihydroxy-2,3,4-trimethoxyxanthone (12), 1,7- dihydroxy-3,4-dimethoxyxanthone (13), 1,7-dihydroxy-3-methoxyxanthone (14), 1-hydroxy-2,3,4,5-tetramethoxyxanthone (15), 1-hydroxy-2,3,4,7-tetramethoxyxanthone (16), 1-hydroxy-2,3,5-trimethoxyxanthone (17), 1-hydroxy-2,3,7-trimethoxyxanthone (18), sweroside (19), gentiopicrin (20), swertiamarin (21), deacetylcentapicrin (22), amaronitidin (23), and amarogentin (24). Conclusion Compound 1 is a new compound named 2-methoveratriloside, and compounds 2, 3, 5, 6, 8, 10-12, 14, and 22-24 are isolated from the Veratrilla genus for the first time.
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Amarogentin is an efficacious Chinese herbal medicine and a component of the bitter apricot kernel. It is commonly used as an expectorant and supplementary anti-cancer drug. β-Glucosidase is an enzyme that hydrolyzes the glycosidic bond between aryl and saccharide groups to release glucose. Upon their interaction, β-glucosidase catalyzes amarogentin to produce considerable amounts of hydrocyanic acid, which inhibits cytochrome C oxidase, the terminal enzyme in the mitochondrial respiration chain, and suspends adenosine triphosphate synthesis, resulting in cell death. Hydrocyanic acid is a cell-cycle-stage-nonspecific agent that kills cancer cells. Thus, β-glucosidase can be coupled with a tumor-specific monoclonal antibody. β-Glucosidase can combine with cancer-cell-surface antigens and specifically convert amarogentin to an active drug that acts on cancer cells and the surrounding antibodies to achieve a killing effect. β-Glucosidase is injected intravenously and recognizes cancer-cell-surface antigens with the help of an antibody. The prodrug amarogentin is infused after β-glucosidase has reached the target position. Coupling of cell membrane peptides with β-glucosidase allows the enzyme to penetrate capillary endothelial cells and clear extracellular deep solid tumors to kill the cells therein. The Chinese medicine amarogentin and β-glucosidase will become an important treatment for various tumors when an appropriate monoclonal antibody is developed.
Subject(s)
Humans , Amygdalin , Therapeutic Uses , Antibodies, Monoclonal , Therapeutic Uses , Antineoplastic Agents , Therapeutic Uses , Cell-Penetrating Peptides , Therapeutic Uses , Iridoids , Therapeutic Uses , Prodrugs , Therapeutic Uses , beta-Glucosidase , Therapeutic UsesABSTRACT
Objective: To investigate the chemical constituents of Swertia chirayita. Methods: Column chromatography, such as silica gel, MCI, Sephadex LH-20 were used to isolate and purify the compounds. Physicochemical properties and spectroscopic methods were used to elucidate their structures. Results: Twelve compounds, including 2 xanthones, 4 triterpenoids, 3 secoiridoids, and 3 other compounds, the chemical constituents were isolated from the ethyl acetate fraction from 85% ethanol extract of S. chirayita, and identified as bellidifolin (1), norbellidifolin (2), oleanolic acid (3), 4-epi-hederagenin (4), 2-epi-corosolic acid (5), ursolic acid (6), amarogentin (7), swerimilegenin I (8), erythrocentaurin (9), pyrocatechol (10), syringic acid (11), and 4-hydroxy-3-methoxybenzoic acid (12). Conclusion: Compounds 4, 5, and 11 are isolated from genus Swertia for the first time, compounds 8 and 9 are found from S. chirayita for the first time.
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OBJECTIVE: To explore the effect of amarogentin on the induction of liver cancer cell line Huh-7 apoptosis and the regulation of PKA/C. METHODS: Liver cancer cell line Huh-7 were divided into 4 groups, control, amarogentin, amarogentin+H89 and amarogentin+H7 group (n=8). The cells were treated with amarogentin (30 mmolL-1) for 6 h besides control group. The amarogentin+H89 and amarogentin+H7 group cells were treated with corresponding compounds at the last 3 h (H89 at 10 mmolL-1 and H7 10 mmolL-1). The apoptotic proteins and MEK/ERK signaling pathway related proteins were detected by Western blotting. The Caspase 3 and Caspase 9 were also be assayed by immune-cytochemistry. At the meaning time, the apoptosis state was assayed by DAPI. RESULTS: The results showed that the Bax, Caspase 3 and Caspase 9 were increased (P>0.05) while the Bcl2 were decreased (P>0.05) expressed greatly after the medication of amarogentin when compared with control. At the same time, the expression of Ras, Rsf, MEK and ERK1/2 were increased (P>0.05) greatly after the medication of amarogentin when compared with control. Those abnormalities were normalized greatly by the medication of H89 (P>0.05) but not H7(P>0.05). CONCLUSION: Amarogentin could promote the apoptosis of liver cancer cell line Huh-7 which is mediated by PKA.
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Objective To improve quality standard of Fufang Shenghua granules.Methods TLC was used to identify chief components in the preparation, Radix et Rhizoma Glycyrrhizae and Salvia Miltiorrhiza.HPLC was applied to identify Amarogentin and to determine the content of Salvianolic acid B.Salvianolic acid B assay was performed on Agilent HC-C18(4.6 mm×250 mm, 5 μm) column with Acetonitrile-0.1% phosphoric acid (23∶77)as mobile phase.The flow rate was 1.0 ml/min.The column temperature was 30 ℃.The detection wavelength was set at 286 nm.Results The spots on TLC were fairly clear with good separation.There was no interference from the negative control samples.However, HPLC was a more accurate, reliable and objective method for qualitative identification.Salvianolic acid B showed a good linear correlation in the range of 1.56~49.92 μg/ml (r=0.999 9).The average recovery was 100.07%, RSD 1.61% (n=9).Conclusion A simple, accurate and reliable method was developed for the quality control of Fufang Shenghua granules.