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ObjectiveTo analyze and predict the potential quality markers （Q-Marker） in the Genuine medicinal materials Jiangxi Aurantii Fructus based on fingerprints and network pharmacology. MethodUltra-high performance liquid chromatography （UPLC） and gas chromatography-mass spectrometry （GC-MS） fingerprints were established for 18 batches of Jiangxi Aurantii Fructus ，combined with chemometric methods to screen out candidate Q-Marker components.Use network pharmacology to construct a "core component-target-pathway" network to predict the Q-Marker and core targets of Jiangxi Aurantii Fructus，and then verify the biological activity of Jiangxi Aurantii Fructus Q-Marker by molecular docking method. ResultThe 18 batches of Jiangxi Aurantii Fructus use UPLC，GC-MS fingerprints combined with chemometric analysis，a total of 9 Q-Marker candidate components were screened out.Through network pharmacological analysis，it is predicted that nobiletin，neohesperidin，meranzin，naringin and D-limonene are the Q-Marker of Jiangxi Aurantii Fructus，acting on the core targets transforming protein p21/H-Ras-1（HRAS），cellular tumor antigen p53 （TP53），mitogen-activated protein kinase 8 （MAPK8），transcription factor AP-1（JUN），glycogen synthase kinase-3 beta（GSK3B），tumor necrosis factor（TNF），cyclin-dependent kinase inhibitor 1（CDKN1A），cAMP-dependent protein kinase catalytic subunit alpha（PRKACA），cysteine aspartate-specific protease-9（Caspase-9），cyclic AMP-responsive element-binding protein 1（CREB1），exerting gastrointestinal motility and antidepressant，anti-inflammatory，anti-tumor，etc.； molecular docking shows that nobiletin，neohesperidin，meranzin，naringin and D-limonene and the selected 10 core targets have good binding ability，reflecting the better biological activity of the Q-Marker of Jiangxi Aurantii Fructus. ConclusionThe Q-Marker of Jiangxi Aurantii Fructus can be comprehensively predicted from the two aspects of volatile and non-volatile components，providing a reference for the quality control of Jiangxi Aurantii Fructus and the further study of its pharmacodynamic mechanism.
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This study was designed to predict the Q-markers of Citri Reticulatae Pericarpium volatile oil and conduct quantitative analysis by GC-MS. The common components of Citri Reticulatae Pericarpium volatile oil were detected by GC-MS. The network pharmacology approaches were utilized for constructing the component-target network and protein-protein interaction(PPI) network, followed by the GO and KEGG pathway enrichment analysis to clarify the pharmacological effects of common components. Molecular docking was conducted to observe the biological activities of common components, thus identifying the Q-markers of Citri Reticulatae Pericarpium volatile oil. The obtained Q-markers were subjected to quantitative analysis by GC-MS. The GC-MS analysis of 19 batches of Citri Reticulatae Pericarpium volatile oil revealed three common components, namely, D-limonene, γ-terpinene, and myrcene. The common components were analyzed based on network pharmacology, and the results showed that Citri Reticulatae Pericarpium volatile oil mainly acted on the core targets GABRA1, GABRA6, GABRA5, GABRA3, and GABRA2 through D-limonene and γ-terpinene, with five important pathways such as nicotine addiction and GABAergic synapse involved. The core targets were mainly distributed in olfactory region, cerebral cortex, cerebellum, basal ganglia, hippocampus, and amygdala to exert the pharmacological effects. As revealed by molecular docking, D-limonene and γ-terpinene exhibited good biological activities, so they were identified as the Q-markers of Citri Reticulatae Pericarpium volatile oil. The results of quantitative analysis showed that the volume fraction of D-limonene was within the range of 0.77-1.03 μL·mL~(-1), and that of γ-terpinene within the range of 0.04-0.13 μL·mL~(-1). The prediction of D-limonene and γ-terpinene as the Q-markers of Citri Reticulatae Pericarpium volatile oil has laid an experimental foundation for the establishment of the quality evaluation standard for Citri Reticulatae Pericarpium volatile oil.
Subject(s)
Citrus , Drugs, Chinese Herbal/pharmacology , Gas Chromatography-Mass Spectrometry , Molecular Docking Simulation , Network Pharmacology , Oils, Volatile/pharmacologyABSTRACT
Objective:To compare volatile components in raw products and steamed products of Citri Sarcodactylis Fructus. Method:Volatile oil in different batch of raw products and steamed products of Citri Sarcodactylis Fructus was extracted by steam distillation, and volatile components were identified by GC-MS.The determination was performed with injector temperature of 280℃, high pure helium(≥ 99.999%) as carrier gas, split ratio of 10:1, and sample size of 3 μL with temperature programming.Mass spectrum conditions included electron bombardment ion source, electron bombardment energy of 70 eV, ion source temperature of 230℃, acceleration voltage of 34.6 V, resolution of 2 500, scanning range of m/z 40-350.Peak matching, cutting and noise filtering were used in analyzing data based on GC-MS combined with orthogonal partial least square-discriminant analysis(OPLS-DA). Result:A total of 58 components were separated and identified from the different batches of raw Citri Sarcodactylis Fructus and their processed products, and 27 differential chemical components were identified by multivariate statistical analysis, among them, 15 common differential components presented different changing laws.The relative contents of α-pinene, β-pinene, p-cymene, β-linalool, L-4-terpinenol, α-terpineol, nerol, β-cyclocitral, geraniol and α-citral in raw products were higher than that of their corresponding processed products.The relative contents of β-caryophyllene, cis-α-bergamotene, γ-muurolene, γ-elemene and β-bisabolene in processed products were higher than that of their corresponding raw products. Conclusion:The content and composition of volatile components in Citri Sarcodactylis Fructus from Guangdong province have changed after being steamed.The results can provide experimental basis for expounding the processing principle and quality evaluation of Citri Sarcodactylis Fructus, and have positive significance for promoting the research and development of this Lingnan characteristic decoction pieces.
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Objective: To preliminarily ascertain the anti-gastric cancer active parts from the roots of Ferula ferulaeoide and to study its GC-MS fingerprint Methods: The inhibitory effects of different extraction from the roots of F. ferulaeoide on the cell proliferation of SGC-7901 cells were determined with MTT colorimetric method. GC-MS fingerprint of in vitro anti-gastric cancer active parts from F. ferulaeoide was investigated and analyzed by GC-MS and principal component analysis (PCA). Results: The chloroform extract showed the best inhibition on the growth of SGC-7901 cells. The method on GC-MS fingerprint of in vitro anti-gastric cancer active parts from F. ferulaeoide was established, showing 28 common characteristic peaks. The PCA demonstrated that the common peaks 1, 2, 4, 6, 7, 8, 9, 12, 21, 22, and 23 were connected closely with the in vitro anti-gastric cancer activity, and the seven compounds were 3-methoxy-1,2-propanediol, D-limonene, L-borneol acetate, terpinyl acetate, 1,5,9-undecatriene, 2,6,10-trimethyl, α-cedrene, and a-bergsmotene, β-cedrene, 8-epi-γ-eudesmol, γ-eudesmol, hinesol. Totally 28 common compounds of 10 batches of samples accounted for over 92% of the volatile contents, and the similarity of the GC-MS fingerprints from the 10 batches of samples was over 0.90. Conclusion: The chloroform extract from the roots of F. ferulaeoides with potential inhibitory effect on gastric cancer SGC-7901 cells is tentatively confirmed, and needs further to verify by animal cells in vivo. The method of GC-MS fingerprinting is rapid, simple, and accurate with good reproducibility and stability, and can be used to control the quality of active parts of F. ferulaeoides.
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Objective To investigate the effects of D-limonene on the proliferation and ras P21 expression in human bladder carcinoma EJ cell.Methods The cell proliferation was measured by XTT assay.The expression of ras mRNA was detected by real-time PCR,and the expression of its protein product was analyzed by Western Blot.Results The proliferation of EJ cell was inhibited by exposure to 2 mmol/L and 4 mmol/L D-limonene for 48h.The optical density (A) was 0.801±0.016 and 0.148±0.008,respectively.There were significant differences between the experimental groups and the control group (1.2181±0.031,P<0.05).The expressions of ras P21 and its protein product of EJ cell were decreased obviously when exposure to 1 mmol/L of D-limonene for 48h.The expres-sions were significantly different compared with the control group(P<0.05).Conclusions D-limo-nene could inhibit human bladder carcinoma EJ cell proliferation,which is in a dose-dependant man-ner.In a lower dose (<1 mmol/L),D-limonene can decrease the expression of ras mRNA and its protein product.
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ObjectiveTo observe the effects of D-limonene on lymphangiogenesis and lymph node metastasis in breast cancer. Methods Human breast cancer cell lines MDA-MB-435 were implanted into the mammary fat pad of 24 female nude mice to establish breast cancer model. The tumor volume and the incidences of lymph node metastasis were detected. Immunohistochemical staining was used to detect the lymphatic microvessel density and VEGF-C expression of the breast cancer tissues. Results In D-limonene group, tumor volume(0.824?0.31) and incidences of lymph node metastasis(25.0%) were significantly inhibited compared with control group (2.178?0.35,87.5%)(all P