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OBJECTIVE@#To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.@*METHODS@#MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.@*RESULTS@#The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.@*CONCLUSION@#DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
Subject(s)
Humans , K562 Cells , Patrinia , Methylene Chloride/pharmacology , Apoptosis , Cell Proliferation , Cell DifferentiationABSTRACT
Objective: To establish the HPLC fingerprint of herbs of Patriniae Herba. Methods: The fingerprints of Patriniae Herba were built using Orca C18 column and acetonitrile-0.05% phosphoric acid as mobile phase. The flow rate was 1.0 mL/min. The detecting wavelength was set at 230 nm. The temperature of column was at 35℃. Results: Under the selected spectrum conditions, HPLC fingerprint of herbs of Patriniae Herba was established, and eight public peaks were shown in the HPLC fingerprint. The methodological study met the required standards. Cluster analysis showed that class I medical BJ1, BJ2, BJ3, BJ4, BJ5, BJ6, BJ7, BJ8, BJ9, both, BJ13, BJ14 each batch Patriniae Herba and control the similarity between fingerprints was 0.987-0.907, Indicating that there are good consistency between batches of medicinal materials; class II medical BJ11, BJ12, and BJ15 group of Patriniae Herba and control fingerprint differences larger. Conclusion: The method is accurate and reliable, simple and efficient, and can be used as the evaluation of Patriniae Herba from different origins and with different varieties.
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OBJECTIVE: To extract and analyze the chemical constituents from Patrinia scabiosaefolia Fisch investigate the antibacterial activities and minimum inhibitory concentration (MIC) of the chemical constituents. METHODS: The chemical constituents from Patrinia scabiosaefolia Fisch were extracted by SFE-CO2 and analyzed by GC-MS. The antimicrobial activities of the chemical constituents were tested by the agar plate diffusion method. The MIC was obtained by microplate method. RESULTS: A total of twenty-four constituents representing 95.407% of the total constituents were identified. The chemical components showed promising antibacterial activity against Staphylococcus aureus, Shigella flexneri and Salmonella spp. The MIC for Staphyloccocus aureus, Shigella flexneri is 4 mg·L-1, and for Salmonella spp, the MIC is 5 mg·L-1. CONCLUSION: The experiment provides a scientific basis for further development and utilization of Patrinia scabiosaefolia Fisch.
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Objective: To study the chemical constituents of Patrinia scabiosaefolia. Methods: The constituents were extracted by methanol and isolated by silica gel chromatography, Sephadex LH-20 gel column chromatography, medium pressure column chromatography, and semi-preparative high performance liquid chromatography. Their structures were elucidated by chemical properties and spectroscopic analysis. Results: Fourteen compounds were isolated and their structures were identified to be 3-hydroxy-olean-11-oxo-12-en-28-oic acid (1), 3, 11-dioxoolean-12-en-28-oic acid (2), 3-O-β-D-xylopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl ester (3), 29-hydroxy-3-oxo-olean-12-en-28-oic acid (4), 3β, 12α-dihydroxy-oleanan-13β, 28-olide (5), 3-O-rhamnopanosyl-(1→2)-xylopyanosyl-oleanolic acid-28-O-glucopyanosyl ester (6), guaia-6-en-4, 10-diol (7), 3-epi-ursoloic acid (8), stigmasterol (9), ergost-6, 22-dien-3β, 5α, 8α-triol (10), oleanolic acid 3-O-β-D-xylopyranoside (11), oleanolic acid 3-α-L-rhamnopyanosyl- (1→2)-β-D-xylopyranoside (12), 3-O-β-D-xylopyanosyl-(1→3)- α-L-rahmnopyanosyl-(1→2)-α-L-arabinopyanosyl oleanolic acid 28-O-β-D-glucopyranosyl ester (13), and oleanolic acid 28-O-β-D- glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (14). Conclusion: Compounds 1-8 are obtained from the plants of Patrinia Juss. for the first time.