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Objective: To explore the network regulation mechanism of blood-activating and hemostatic and detumescent and analgesic traditional effects of Panax notoginseng. Methods: Targets of the 12 components of P. notoginseng absorbed in plasma were predicted according to the reverse pharmacophore method. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by Omicsbean online analysis software and String 10 database. Finally, Cytoscape software was used to construct the network pharmacology map. Results: A total of 12 compounds (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rh1, ginsenoside Rg2, ginsenoside Rb1, ginsenoside Rd, ginsenoside F2, ginsenoside Rg3, ginsenoside Rk1, dencichine and quercetin) affected 65 pathways through 65 related targets, which were associated with anti-thrombosis, fibrinolysis, angiogenesis, vasodilation, blood coagulation, anti-inflammation and analgesia. The network of "compound-target-pathway-pharmacological action-efficacy" was also constructed. Conclusion: P. notoginseng interferes with multiple biological processes related to activating blood circulation, hemostasis, detumescence and analgesia by acting on several key proteins such as F2, F10, PLAT, VEGFA, NOS2, IL6, PTGES, OPRD1, etc.
ABSTRACT
Objective: Through network pharmacology, the network relationship between the active component of Sanqi Mixture, the target of hepatic ischemia- reperfusion injury(HIRI), and biological pathway was constructed to explore the key target and mechanism of effect of Sanqi Mixture on HIRI. Method: Through literature research at home and abroad, Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform, Pharm Mapper, Swiss Target Prediction and other servers, oral availability (OB) and drug-likeness (DL) were selected as the limited conditions to collect the relevant targets for Sanqi Mixture for intervention in HIRI. The OMIM database was used to screen and collate HIRI related genes and protein targets. Excel table was used to merge and sort the intersection between disease and targets through Cytoscape3.7.2 software plug-ins Network Analyzer, with topological parameters (degree) ≥ 5 (average degrees of freedom 4.5) for the filter to find the core targets; And the intersection targets were imported to the server STRING, and with Confidence Score of 0.85 or higher for the filter conditions to build the core protein interactions (Hub-PPI) network. The intersection target was introduced into FunRich 3.0 software for biological process and biological pathway analysis, and Cytoscape3.7.2 was used to construct the network of "traditional Chinese medicine-active ingredient-HIRI target-biological pathway". Result: Sanqi mixture could reduce the expression of Aspartate aminotransferase (AST) and glutamate transaminase (ALT) in HIRI mice (P < 0.01). After screening, 45 active components of Sanqi Mixture were obtained, corresponding to 3 273 targets, and the main compounds included ursolic acid, oleanolic acid, brucine, quercetin, ginsenoside F2, paeoniflorin, etc. Among the 196 targets obtained by HIRI, 46 targets were intersected with components, including 11-β-hydroxysteroid dehydrogenase (HSD11B1), adenosine receptor A3 (ADORA3), cyclooxygenase 2 (PTGS2), adenosine receptor A1 (ADORA1), protein kinase C-ε (PKC), etc. With the STRING server setting the qualified condition of Confidence Score ≥ 0.85, the PPI network with high Confidence was obtained and clustered into three categories through cluster processing. Five biological processes including protein metabolism, signal transduction, negative regulation of enzyme activity, inflammatory response and transmembrane receptor protein tyrosine kinase signal pathway were analyzed by FunRich software (P < 0.05). 16 biological pathways including integrin-linked kinase signal, TNF receptor signaling pathway, P38 mitogen-activated protein kinase signaling pathway, and TRAIL signaling pathway (P < 0.01). Conclusion: It is preliminarily discussed that Sanqi Mixture intervenes HIRI through the interaction of multiple components and multiple targets, as well as the regulation of multiple biological pathways and biological processes. However, the key core targets and the specific regulation mechanism still need further experimental verification.
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Objective: To establish an HPLC fingerprint of Qinqi Rheumatism Formula (QRF) for its quality control and effective components determination. Methods: HPLC method was performed on Inert Sustain C18 (150 mm × 4.6 mm, 5 μm) column with gradient elution composed of acetonitrile-aqueous solution containing 0.1% phosphoric acid at the flow rate of 1.0 mL/min. The column temperature was set at 30℃, while the detective wavelength was set at 203 nm. The common mode of HPLC fingerprint for 10 batches of QRF was established with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2004 A edition) and the common peaks were identified by reference compounds. Results: Fingerprints of QRF were established. The similarities of the 10 batches of samples were above 0.99. A total of 19 common peaks were found. Eight mutual peaks were from Panacis Majoris Rhizoma, eight mutual peaks were from Gentianae Macrophyllae Radix, and five mutual peaks were from Corni Fructus (Peak 1 was the common peak from Panacis Majoris Rhizoma, Gentianae Macrophyllae Radix, and Corni Fructus). Based on the retention time of reference substances, eight constituents including loganic acid (peak 2), morroniside (peak 3), gentiopicroside (peak 5), loganin (peak 6), ginsenoside Ro (peak 12), ginsenoside F1 (peak 13), panax japonicus IVa (peak 14), and ginsenoside F2 (peak 17) were indentified. Conclusion: The method is stable, specific, and reproducible, and can be used for the quality control of QRF and the study of its effective components.
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Objective: To evaluate the quality of Maiweishen, a simple and accurate HPLC method for determining the contents of 20 active constituents from Maiweishen was established. Methods: The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm, 5 μm) using a mobile phase made up of acetonitrile and water at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set as 203 nm and 35 ℃, respectively. Results: Sixteen ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, compounds K, Rh2, and protopanaxadiol), three kinds of lignan in Schisandra chinensis (schizandrol A, schizandrin A, B), and ophiopogonin D were separated at baseline with good linearity (r ≥ 0.999 6). The recovery rates were 96%-102% (RSD < 2%). Conclusion: The method is simple, fast, accurate, and could be applied to the quality control of Maiweishen.
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Objective: A high-performance liquid chromatography-tandem mass spectrometric method for the simultaneous determination of 15 ginsenoside compounds from Panacis Majoris Rhizoma (PMR) was developed. Methods: A Waters Sunfire ™ C18 column (150 mm × 4.6 mm, 5 μm) was used for the separation. The mobile phase consisted of A (H2O + 0.05% HCOOH) and B (CH3CN + 0.05% HCOOH) using a gradient elution. For the quantification of ginsenosides, the multiple reaction monitoring (MRM) mode of the mass spectrometer was applied and the declustering potential (DP), collision energy (CE), and collision cell exit potential (CXP) were optimized to perform automatic on-line MS/MS experiments during the chromatographic separation. Results: By using the optimized method, the linearity range of 15 analytes was 0.000 9 to 2 952.592 3 μg/mL with more than 0.999 determination coefficient (r) of linear regressions, the detection limits of the 15 ginsenosides ranged from 0.003 to 626.554 ng/mL, the limits of quantitation ranged from 0.075 to 1 762.150 ng/mL, the recoveries of 15 ginsenosides in the samples were 98.15%-101.12% with relative standard deviation (RSD) that ranged from 0.82% to 2.15%. Conclusion: The proposed LC-MS/MS method is accurate and reproducible in accordance with TCM guidelines, showing high sensitivity, rapidness, and recovery. This method allows the assessment of various ginsenosides in a single analytical run providing an innovative tool to control Panacis Majoris Rhizoma materials quantification.