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OBJECTIVE@#To determine whether monotropein has an anticancer effect and explore its potential mechanisms against colorectal cancer (CRC) through network pharmacology and molecular docking combined with experimental verification.@*METHODS@#Network pharmacology and molecular docking were used to predict potential targets of monotropein against CRC. Cell counting kit assay, plate monoclonal assay and microscopic observation were used to investigate the antiproliferative effects of monotropein on CRC cells HCT116, HT29 and LoVo. Flow cytometry and scratch assay were used to analyze apoptosis and cell cycle, as well as cell migration, respectively in HCT116, HT29, and LoVo cells. Western blotting was used to detect the expression of proteins related to apoptosis, cell cycle, and cell migration, and the expression of proteins key to the Akt pathway.@*RESULTS@#The Gene Ontology and Reactome enrichment analyses indicated that the anticancer potential of monotropein against CRC might be involved in multiple cancer-related signaling pathways. Among these pathways, RAC-beta serine/threonine-protein kinase (Akt1, Akt2), cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-9 (MMP9), epidermal growth factor receptor (EGFR), cell division control protein 42 homolog (CDC42) were shown as the potential anticancer targets of monotropein against CRC. Molecular docking suggested that monotropein may interact with the 6 targets (Akt1, Akt2, CDK6, MMP9, EGFR, CDC42). Subsequently, cell activity of HCT116, HT29 and LoVo cell lines were significantly suppressed by monotropein (P<0.05). Furthermore, our research revealed that monotropein induced cell apoptosis by inhibiting Bcl-2 and increasing Bax, induced G1-S cycle arrest in colorectal cancer by decreasing the expressions of CyclinD1, CDK4 and CDK6, inhibited cell migration by suppressing the expressions of CDC42 and MMP9 (P<0.05), and might play an anticancer role through Akt signaling pathway.@*CONCLUSION@#Monotropein exerts its antitumor effects primarily by arresting the cell cycle, causing cell apoptosis, and inhibiting cell migration. This indicates a high potential for developing novel medication for treating CRC.
Subject(s)
Humans , Proto-Oncogene Proteins c-akt/metabolism , Cell Proliferation , Matrix Metalloproteinase 9 , Molecular Docking Simulation , Cell Cycle , ErbB Receptors , Apoptosis , Colorectal Neoplasms/pathology , Cell Line, TumorABSTRACT
Objective: A method for identification of root cortex and woody core of Morindae Officinalis Radix (MOR) was established based on Ultra Performance Liquid Chromatography (UPLC) characteristic chromatogram and chemical pattern recognition technique. Methods: Using UPLC technique, the characteristic chromatograms of iridoids and oligosaccharides of root cortex and woody core of MOR were established, combined with the similarity analysis, variance analysis, cluster analysis, principal component analysis (PCA) methods for chemical pattern recognition research. Results: The UPLC characteristic chromatograms of iridoids and oligosaccharides of different parts of MOR were established, and 12 and 20 common characteristic peaks were confirmed, respectively. The UPLC characteristic chromatograms of root cortex and woody core of MOR were obviously different. Conclusion: UPLC characteristic chromatograms of iridoids and oligosaccharides of MOR combined with chemical pattern recognition analysis method can reflect the difference of root cortex and woody core of MOR integrally, comprehensively and truly, which provides more sufficient basis for the necessity of removing woody core from MOR.
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Objective To optimize the extraction process of main iridoid glycosides in Damnacanthus officinarum Huang and evaluate the antioxidant activity of Damnacanthus officinarum Huang in vitro. Methods The classical heating-reflux extraction method was selected. The volume fraction of ethanol, the volume of solvent and extraction time were taken as the evaluation factors. The comprehensive score of extraction yield and the monotropein content were used as the evaluation indexes. An orthogonal test was designed to select the best extraction conditions. The total reducing capacity, DPPH clearance rate and hydroxyl radical scavenging rate were measured to determine its antioxidant activity in vitro. Results The optimal extraction process was the reflux with 6 times volume of 60% ethanol for 2 hours. Damnacanthus officinarum Huang has certain antioxidant capacity, and the activity of ethyl acetate part had the best effect. Conclusion The optimized extraction process is stable and feasible, which can be used for extraction of the iridoid glycosides from Damnacanthus officinarum Huang. This study has proved that Damnacanthus officinarum Huang has certain antioxidant activity.
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Abstract A selective and sensitive liquid chromatography tandem with mass spectrometry was developed and validated for accurate determination of monotropein in rat plasma and tissues. All biological samples were prepared by simple protein precipitation method using catalpol as an internal standard. The analyte and internal standard were separated on a C18 analytical column with 2 min of run time, at flow rate of 0.5 ml/min. The detection was performed on a triple-quadrupole tandem mass spectrometer equipped with negative-ion electrospray ionization by selected-reaction monitoring of the transitions at m/z 389→147 for monotropein and m/z 361→169 for the internal standard. The calibration curves for plasma and tissue samples were linear over the concentration range of 4-2000 ng/ml, with a lower limit of quantification of 4 ng/ml. The method was successfully applied to a pharmacokinetic and tissue distribution study of monotropein in rats.
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Objective To establish qualitative and quantitative methods for analyzing monotropein in Liquidambaris Radix. Methods Monotropein in Liquidambaris Radix was identified by using TLC. The contents of monotropein in the samples were determined by HPLC on an Sino Chrom ODS-BP column (4.6 mm × 250 mm, 5 μm) with methanol-0.1％ phosphoric acid solution (1:99) as the mobile phase at the flow rate of 1.0 m L/min; the detection wavelength was 235 nm; the column temperature was 25 ℃; the injection volume was 10 μL. Results Monotropein in Liquidambaris Radix could be identified by TLC. Monotropein showed good linear relation in the range of 0.150 1– 1.501 0 μg (r=0.999 8), and the average recovery rate was 95.93％ (RSD=0.60％). The contents of monotropein in Liquidambaris Radix from 10 different producing areas were among the range of 0.20％–1.15％. Conclusion The method is simple, stable, reliable and reproducible, which can be used for the quality control of Liquidambaris Radix.
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Objective:To optimize the processing technology of processed Morinda Officinalis by taking monotropein content as the index. Methods:The amount of licorice,stir frying time and cooking pot temperature were used as the investigation factors in orthogonal experiments design,and an HPLC method was used to determine the content with he following chromatographic conditions:Kromalsil C18 chromatographic column(200 mm × 4. 6 mm,5μm),mobile phase of methanol-0. 4% phosphoric acid solution(90 ∶10),the column tem-perature at 30℃,the flow rate at 1 ml·min-1 ,the detection wavelength at 233 nm and the sample size of 5μl. Results:The monotropein content of each processed Morinda Officinalis sample after different processing was:0. 535 6, 0. 582 4, 0. 523 4, 0. 589 1, 0. 578 6, 0. 587 8,0. 575 2,0. 609 1 and 0. 558 7 mg·g-1,which showed that the processing technology could affect,the monotropein content. Conclusion:The best processing technology of preparation Morinda Officinalis based on Monotropein content is: Liquorice of 6%,stir time of 10min and boiling pot temperature at 100 ℃.
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Objective To establish a method of quality control for compound Cornu cervi degelatinatum tablets. Methods Thin layer chromatography ( TLC ) was used for qualitative identification of Lonicerae Japonicae Caulis and Pyrola Herba in the preparations. High performance liquid chromatography was used to determine the content of loganin and chiratin in Lonicerae Japonicae Caulis and monotropein in Pyrola Herba. Results The TLC spots were clear and specific in the qualitative identification.HPLC determined the content. In the concentration range, monotropein, loganin and chiratin had a good linear relationship with peak area (r>0.999).The mean recovery was 97.2%, 98.4% and 96.0%;RSD was 1.4%, 0.8% and 0.8%. Conclusion The present study employs TLC for quality control and HPLC for quantity control. The methods are simple and accurate, have good reproducibility, and can effectively control the quality of compound Cornu cervi degelatinatum tablets.
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Objective To determine the content of monotropein in the different parts of Morinda officinalis and in its counterfeit species.Methods The method of HPLC was used with chromatographic conditions as follows:Kromasil C18(150 mm?4.6 mm,5 ?m) column,the mobile phase of methol-0.4 %phosphate solution(5:95→28.8:71.2,15 min),the velocity of flow being 1 mL/min,the detection wavelength at 231 nm and column temperature being 25 ℃.Results The content of monotropein in the leaves and radix of Morinda officinalis and Morinda parvifolis is the highest.Conclusion It is reasonable to replace the root of Morinda officinalis with the leaves of Morinda officinalis and Morinda parvifolis for the monotropein extraction.