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OBJECTIVE@#To explore the effects of isobavachalcone (IBC) on cell death of human breast cancer MCF-7 cells and explore the possible mechanism.@*METHODS@#MCF-7 cells were treated with different concentrations of IBC, and the changes in cell proliferation were assessed using MTT assay. Apoptosis of MCF-7 cells following treatment with 10, 20, and 40 μmol/L IBC was analyzed using flow cytometry with annexin V-FITC/PI double staining and fluorescence microscopy, and the expressions of apoptosis- and autophagy-related proteins (Bax, Bcl-2, Akt, p-Akt, p62, and LC3) were detected with Western blotting. Electron microscopy was used to observe the changes in submicrostructure of the cells following treatment with 40 μmol/L IBC. JC-1 assay kit, ATP assay kit, and reactive oxygen species (ROS) kit were used to determine the effect of IBC on mitochondrial function of the cells.@*RESULTS@#MTT assay showed that IBC significantly inhibited the proliferation of MCF-7 cells in a concentration- and time-dependent manner, with IC50 values of 38.46, 31.31, and 28.26 μmol/L at 24, 48, and 72 h, respectively. IBC also concentration-dependently induced apoptosis of MCF-7 cells. IBC-induced cell death was inhibited by z-VAD-fmk, a caspase inhibitor (P < 0.05), but not by the necroptosis inhibitor necrostatin-1 (Nec-1). Western blotting showed that IBC-induced MCF-7 cell apoptosis by increasing Bax expression and down-regulating the expressions of Bcl-2, Akt and p-Akt-473 (all P < 0.05). With the increase of IBC concentration, the expression of autophagy-related protein p62 and the LC3-II/I ratio increased progressively. Electron microscopy revealed the presence of autophagic bodies in IBC-treated MCF-7 cells. IBC treatment also resulted in decreased mitochondrial membrane potential and intracellular ATP level and increased ROS accumulation in MCF-7 cells (P < 0.05).@*CONCLUSION@#IBC is capable of inducing both apoptosis and autophagy in MCF-7 cells, suggesting the potential value of IBC as a lead compound in the development of anti-breast cancer agents.
Subject(s)
Humans , Adenosine Triphosphate , Cell Death , Chalcones , MCF-7 Cells , Neoplasms , Proto-Oncogene Proteins c-akt , Proto-Oncogene Proteins c-bcl-2/metabolism , Reactive Oxygen Species/metabolism , bcl-2-Associated X ProteinABSTRACT
Pancreatic lipase (PL), a crucial enzyme in the digestive system of mammals, has been proven as a therapeutic target to prevent and treat obesity. The purpose of this study is to evaluate and characterize the PL inhibition activities of the major constituents from Fructus Psoraleae (FP), one of the most frequently used Chinese herbs with lipid-lowering activity. To this end, a total of eleven major constituents isolated from Fructus Psoraleae have been obtained and their inhibition potentials against PL have been assayed by a fluorescence-based assay. Among all tested compounds, isobavachalcone, bavachalcone and corylifol A displayed strong inhibition on PL (IC < 10 μmol·L). Inhibition kinetic analyses demonstrated that isobavachalcone, bavachalcone and corylifol A acted as mixed inhibitors against PL-mediated 4-methylumbelliferyl oleate (4-MUO) hydrolysis, with the K values of 1.61, 3.77 and 10.16 μmol·L, respectively. Furthermore, docking simulations indicated that two chalcones (isobavachalcone and bavachalcone) could interact with the key residues located in the catalytic cavity of PL via hydrogen binding and hydrophobic interactions. Collectively, these finding provided solid evidence to support that Fructus Psoraleae contained bioactive compounds with lipid-lowering effects via targeting PL, and also suggested that the chalcones in Fructus Psoraleae could be used as ideal leading compounds to develop novel PL inhibitors.
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Objective: To investigate the chemical compounds from the twigs of Morus multicaulis. Methods: The chemical constituents were isolated by various chromatographic methods from 90% ethanol aqueous extract of M. multicaulis and the structures were elucidated by a combined analysis of physicochemical properties, as well as spectroscopic methods including UV, IR, MS ECD and NMR. Results: Six flavonoids were isolated and the structures were identified as moritwigsone A (1), carpachromene (2), isobavachalcone (3), isoliquiritigenin (4), apigenin (5), and quercitrin (6). The absolute configuration of compound 1 was determined by electronic circular dichroism (ECD) calculation. Conclusion: Compound 1 is a new compound, named moritwigsone A, and compounds 2-4 are isolated from this plant for the first time.
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Objective: To establish UPLC method for the simultaneous determination of 10 components, such as psoralen, isopsoralen, psoralidin, bavachinin, isobavaenin, corylifolin, isobavachalcone, corylin, neobavaisoflavone, and bakuchiol in Psoralea Fructus, and to study the effect of processing time on 10 components in stir-frying Psoralea Fructus with salt solution. Methods: The chromatographic separation was achieved on a C18 column (50 mm×4.6 mm, 1.8 μm) with acetonitrile (A)-water (B) as mobile phase at the flow rate of 1.0 mL/min for gradient elution; The column temperature was 30℃. The determination wavelength was 250 nm. Results: The 10 components were well separated within 15 min. The RSD values of reproducibility were less than 3%. The stability was good in 24 h. The linear relationship between the concentration and peak areas of the 10 components was good (r≥0.9970). The average recoveries were 96%-105% and the RSD values were all less than 3%. Conclusion: The method is simple, reliable and accurate, could be used for the quality control of Psoralea Fructus. With processing time prolonged, the content of coumarins showed first decreased, then increased and last decreased. Flavonoids, bakuchiol and the total content of 10 components above were decreased.
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Objective:To observe the effects of isobavachalcone (IBC)on the proliferation and apoptosis of tongue squamous cell carcinoma Tca-8113 cells,and to explore their mechanisms. Methods:The Tca8113 cells cultured in vitro were divided into control group and different doses (0, 10, 20,40, 80 μmol · L-1 )of IBC groups.The inhibitory rates of cell proliferation were detected by MTT method.The apoptosis was detected by flow cytometry.Western blotting method was used to detect the expressions of Akt,p-Akt,Erk,p-Erk,Bax, Bcl-2 and Caspase-3 proteins in the Tca8113 cells in various groups.Results:The MTT results showed that the inhibitory rates of proliferation of Tca8113 cells were increased in a concentration-and time-dependent manner;the IC50 at 12,24 and 48 h were (285.13±8.97), (132.40±7.76),and (58.56±5.93)μmol·L-1 ,respectively;and there were significant differences between different time points (P 0.05 ).The expression levels of Caspase-3 in Tca8113 cells in 40 μmol·L-1 IBC group at 48 h were increased compared with control group (P <0.05).Conclusion:IBC could inhibit the proliferation and induce the apoptosis of Tca8113 cells;Akt and Erk signaling pathway may be the pathway of IBC to induce the apoptosis of tumor cells.
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OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.
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Aim To explore the inhibitory effect of isobavachalcone (IBC)on migration and invasion of tongue squamous cell carcinoma Tca81 1 3 cells and its possible mechanism.Methods Tca81 1 3 cells were treated in different concentrations of IBC in vitro.Cell proliferation was detected by MTT;Wound healing as-say and Transwell chamber assay were used to detect the ability of cell migration and invasion;Western blot was applied to detect the expression of Akt,p-Akt, MMP-2 and MMP-9 proteins.Results IBC could in-hibit the proliferation of Tca81 1 3 cells in a concentra-tion-and time-dependent manner.IBC can reduce cell migration and invasion.Western blot showed that IBC could an decrease the expression of p-Akt,MMP-2 and MMP-9 proteins in a concentration-dependent manner. However,the level of Akt was not affected by the con-centration of IBC treatment.Conclusion IBC could inhibit the proliferation, migration and invasion in Tca81 1 3 cells and its mechanism may be associated with the down-regulation of MMP-2 and MMP-9 pro-teins and the inhibition of phosphorylation of upstream Akt.
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Objective: To screen the small molecular compounds with anti-asthma activity from Psoralea corylifolia monomor and explain its regulation mechanism. Methods: Based on the effect of IL-4 in pathological features of masthma, a high throughput screening system was designed by using 4get mice and flow cytometry to inhibit IL-4 production so as to anti-asthma. Results: Compared with DMSO group, isobavachalcone showed high ability of inhibiting IL-4 production in Th2 cells (P < 0.001), down-regulating GATA-3 expression (P < 0.01), and decreasing the level of STAT6 phosphorylation (P < 0.01). Conclusion: Isobavachalcone could inhibit the IL-4 production through down-regulating the GATA-3 expression by controling the level of STAT6 phosphorylation. Isobavachalcone is a potential drug for the treatment of asthma.