"Component-target-efficacy" network analysis and experimental verification of Qingkailing Oral Preparation / 中国中药杂志

This study aims to explore the mechanism of Qingkailing(QKL) Oral Preparation's heat-clearing, detoxifying, mind-tranquilizing effects based on "component-target-efficacy" network. To be specific, the potential targets of the 23 major components in QKL Oral Preparation were predicted by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The target genes were obtained based on UniProt. OmicsBean and STRING 10 were used for Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the targets. Cytoscape 3.8.2 was employed for visualization and construction of "component-target-pathway-pharmacological effect-efficacy" network, followed by molecular docking between the 23 main active components and 15 key targets. Finally, the lipopolysaccharide(LPS)-induced RAW264.7 cells were adopted to verify the anti-inflammatory effect of six monomer components in QKL Oral Preparation. It was found that the 23 compounds affected 33 key signaling pathways through 236 related targets, such as arachidonic acid metabolism, tumor necrosis factor α(TNF-α) signaling pathway, inflammatory mediator regulation of TRP channels, cAMP signaling pathway, cGMP-PKG signaling pathway, Th17 cell differentiation, interleukin-17(IL-17) signaling pathway, neuroactive ligand-receptor intera-ction, calcium signaling pathway, and GABAergic synapse. They were involved in the anti-inflammation, immune regulation, antipyretic effect, and anti-convulsion of the prescription. The "component-target-pathway-pharmacological effect-efficacy" network of QKL Oral Preparation was constructed. Molecular docking showed that the main active components had high binding affinity to the key targets. In vitro cell experiment indicated that the six components in the prescription(hyodeoxycholic acid, baicalin, chlorogenic acid, isochlorogenic acid C, epigoitrin, geniposide) can reduce the expression of nitric oxide(NO), TNF-α, and interleukin-6(IL-6) in cell supernatant(P<0.05). Thus, the above six components may be the key pharmacodynamic substances of QKL Oral Preparation. The major components in QKL Oral Prescription, including hyodeoxycholic acid, baicalin, chlorogenic acid, isochlorogenic acid C, epigoitrin, geniposide, cholic acid, isochlorogenic acid A, and γ-aminobutyric acid, may interfere with multiple biological processes related to inflammation, immune regulation, fever, and convulsion by acting on the key protein targets such as IL-6, TNF, prostaglandin-endoperoxide synthase 2(PTGS2), arachidonate 5-lipoxygenase(ALOX5), vascular cell adhesion molecule 1(VCAM1), nitric oxide synthase 2(NOS2), prostaglandin E2 receptor EP2 subtype(PTGER2), gamma-aminobutyric acid receptor subunit alpha(GABRA), gamma-aminobutyric acid type B receptor subunit 1(GABBR1), and 4-aminobutyrate aminotransferase(ABAT). This study reveals the effective components and mechanism of QKL Oral Prescription.

Identification of Q-markers for Cistanches Herba based on HPLC-Q-TOF-MS/MS and network pharmacology / 中国中药杂志

This study aims to establish a method for analyzing the chemical constituents in Cistanches Herba by high performance liquid chromatography(HPLC) and quadrupole-time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS), and to reveal the pharmacological mechanism based on network pharmacology for mining the quality markers(Q-markers) of Cistanches Herba. The chemical constituents of Cistanche deserticola and C. tubulosa were analyzed via HPLC-Q-TOF-MS/MS. The potential targets and pathways of Cistanches Herba were predicted via SwissTargetPrediction and DAVID. The compound-target-pathway-pharmacological action-efficacy network was constructed via Cytoscape. A total of 47 chemical constituents were identified, involving 95 targets and 56 signaling pathways. We preliminarily elucidated the pharmacological mechanisms of echinacoside, acteoside, isoacteoside, cistanoside F, 2'-acetylacteoside, cistanoside A, campneoside Ⅱ, salidroside, tubuloside B, 6-deoxycatalpol, 8-epi-loganic acid, ajugol, bartsioside, geniposidic acid, and pinoresinol 4-O-β-D-glucopyranoside, and predicted them to be the Q-markers of Cistanches Herba. This study identified the chemical constituents of Cistanches Herba, explained the pharmacological mechanism of the traditional efficacy of Cistanches Herba based on network pharmacology, and introduced the core concept of Q-markers to improve the quality evaluation of Cistanches Herba.

Study on critical quality attributes of Qingjin Huatan Decoction based on serum pharmacochemistry / 中国中药杂志

Qingjin Huatan Decoction is a classic prescription with the effects of clearing heat, moistening lung, resolving phlegm, and relieving cough. In order to explore the critical quality attributes of Qingjin Huatan Decoction, we identified the blood components of Qingjin Huatan Decoction by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS) under the following conditions, chromatographic column: Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm); mobile phase: 0.1% formic acid acetonitrile(A)-0.1% formic acid in water(B); gradient elution; flow rate: 0.2 mL·min~(-1); column temperature: 30 ℃; injection volume: 5 μL. The electrospray ionization(ESI) source was used to collect data in both positive and negative ion modes under the following conditions, capillary voltage: 3 kV for the positive ion mode and 2 kV for the negative ion mode; ion source temperature: 110 ℃; cone voltage: 30 V; cone gas flow rate: 50 L·h~(-1); nitrogen degassing temperature: 350 ℃; degassing volume flow rate: 800 L·h~(-1); scanning range: m/z 50-2 000. In this experiment, a total of 66 related components of Qingjin Huatan Decoction were identified, including 22 prototype components and 44 metabolites. The results of this study preliminarily revealed the pharmacodynamic material basis of Qingjin Huatan Decoction in vivo, which has provided an experimental basis for the determination of quality markers of Qingjin Huatan Decoction and the development of new drugs.

Pharmacodynamic material basis and mechanism of Fufang Yuxingcao Mixture for the treatment of heat-clearing and detoxification based on network pharmacology / 药学学报

We explored the pharmacodynamic material basis and network regulatory mechanism of Fufang Yuxingcao Mixture (FYM) for the treatment of fever and inflammation. Targets of the 25 compounds in FYM were predicted according to the reverse pharmacophore method and TCMSP, UniProt database. Gene ontology (GO) function enrichment and pathway analysis of the targets was analyzed by Omicsbean software and the Kyoto Gene and Genome Encyclopedia (KEGG) database. A "compound-target-pathway-pharmacological action-effect" network was established with Cytoscape 3.6.1 software. The lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model was used to verify the anti-inflammatory effects of FYM and its 10 important components. The network pharmacology experiment showed that 25 compounds affected 97 pathways through 211 targets, of which 15 key targets [including RAC-alpha serine/threonine-protein kinase (AKT1), insulin (INS), vascular endothelial growth factor A (VEGFA), interleukin-6 (IL-6), cellular tumor antigen p53 (TP53), tumor necrosis factor (TNF), transcription factor AP-1 (JUN), caspase-3 (CASP3), matrix metalloproteinase-9 (MMP9), interleukin-8 (IL-8), prostaglandin G/H synthase 2 (PTGS2), proto-oncogene c-Fos (FOS), tyrosine-protein kinase SRC (SRC), c-Jun N-terminal kinase 1 (MAPK8), estrogen receptor 1 (ESR1)] and 46 pathways (including NF-kappa B signaling pathway, Toll-like receptor signaling pathway, MAPK signaling pathway, IL-17 signaling pathway, arachidonic acid metabolism, cAMP signaling pathway, T cell receptor signaling pathway, calcium signaling pathway, inflammatory mediator regulation of TRP channels, chemokine signaling pathway, Th1 and Th2 cell differentiation, natural killer cell mediated cytotoxicity, etc.) were related to anti-inflammatory, antipyretic, immune regulation, and analgesia. In vitro cell experiments showed that FYM and the 10 components (including isoquercitrin, luteoloside, baicalein, wogonin, wogonoside, phillyrin, forsythoside A, chlorogenic acid, isochlorogenic acid A, and sweroside) could significantly reduce the expression of nitric oxide (NO), TNF-α and IL-6 in cell supernatants, indicating that the above 10 components may be the key pharmacodynamic material basis of FYM.

Mechanism of Tanreqing Capsule on treatment of coronavirus disease 2019 based on network pharmacology / 中草药

Objective: To explore the network regulation mechanism of Tanreqing Capsule (TRQC) on the treatment of coronavirus disease 2019 (COVID-19). Methods: Potential targets of the 19 major constituents in TRQC were predicted by the Swiss Target Prediction server and TCMSP database. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by Omicsbean analytic system and String 10 database. Finally, Cytoscape 3.6.1 software was used to construct the network pharmacology map. Results: A total of 19 compounds affected 68 pathways such as IL-17 signaling pathway, T cell receptor signaling pathway, arachidonic acid metabolism, cAMP signaling pathway, PI3K-Akt signaling pathway, influenza A, etc, by acting on 163 related targets, which associated with anti-inflammation, immune regulation, antipyresis, eliminating phlegm, relieving cough and asthma, analgesia, antibacterial and antiviral, and sedation. The network of "compound-target-pathway-pharmacological action-efficacy" was also constructed. Conclusion: The major constituents in TRQC, including scutellarin, baicalin, oroxylin-7-O-glucuronide, chrysin-7-O-glucuronide, forsythin, forsythiaside E, forsythiaside D, chlorogenic acid, caffeic acid, isochlorogenic acid A, ursodeoxycholic acid and chenodeoxycholic acid, may interfere with efficacy-related biological processes associated with antipyresis, eliminating phlegm and removing toxin by acting on key protein like TNF, EGFR, NOS3, PTGS2, IL2, GABBR1, MAPK14, ADRB2, REN, VCAM1, ACHE, PTPRC, etc.

Study on mechanism of anti-inflammation and immunoregulation of Shufeng Jiedu Capsule based on network pharmacology / 中草药

Objective: To explore the network regulation mechanism of anti-inflammation and immunoregulation of Shufeng Jiedu Capsule (SJC). Methods: Targets of the 28 compounds in SJC were predicted according to the reverse pharmacophore method. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by MAS 3.0 software and Kyoto Gene and Genome Encyclopedia (KEGG) database. Finally, Cytoscape software was used to construct the network pharmacology map. Results: A total of 28 compounds affected 112 pathways through 127 targets, of which 41 targets and 22 pathways were associated with anti-inflammation and immunoregulation. The network of compound-target-pathway-pathology process was also constructed. Conclusion: SJC interferes with multiple biological processes related to anti-inflammation and immunoregulation by acting on several key proteins such as HRAS, MAP2K1, AKT1, MME, and PTPN1. The mechanism of SJC was preliminarily revealed.

Study on compatibility rationality of Shufeng Jiedu Capsule based on network pharmacology / 中草药

Objective: To explore the compatibility rationality of Shufeng Jiedu Capsule (SJC) for the treatment of upper respiratory infection (URI). Methods: The potential targets and pathways of 32 compounds selected from ‘heat-clearing and detoxicating group’ (HCDG), ‘relieving exterior syndrome group’ (RESG), and ‘Glycyrrhiza uralensis group (GUG)’ of SJC were predicted by the methods of network pharmacology. The mechanism and compatibility rationality of SJC were analyzed by data integration and analysis. Results: The in silico prediction results showed that 32 compounds of SJC affected 34 related pathways through 94 target proteins which mainly involved with inflammation, lipopolysaccharide and bacterium response, immunoreaction and so on. The three groups of HCDG, RESG, and GUG not only showed common targets and pathways but also had their own emphasis to exert synergistic effects. Conclusion: HCDG plays a therapeutic role by intervening in related physiological process, such as inflammatory response, lipopolysaccharide and bacterium response, defense response and immune response. In addition to providing synergistic effects with HCDG, RESG can also intervene in the process of the sweating and antipyretic through multiple pathways. GUG showed auxiliary therapeutic effect by participating in processes of anti-inflammatory and enhancing the body immunity.

Spectrum-effect relationship of wind-dispersing and exterior syndrome-relieving of Shufeng Jiedu Capsules / 中草药

Objective: To explore the potential effective compounds of ‘wind-dispersing and exterior syndrome-relieving’ of Shufeng Jiedu Capsule (SJC). Methods: The fingerprints of 22 samples with different proportions of SJC were established by LC-MS. The stimulation rates of 22 samples to the M3 receptor were measured by Chinese hamster ovarian cancer cells with high expression of M3 acetylcholine receptor which related to ‘wind-dispersing and exterior syndrome-relieving’. Finally, the spectrum-effect relationship of relative peak areas of characteristic peaks and stimulation rates was analyzed by BP-neural network. Results: According to the established optimal neural network model, 16 characteristic peaks were calculated and found to be significantly correlated with the activity of SJC on the M3 receptor stimulation. Conclusion: Through the spectrum-effect study, it is speculated that the effective components of SJC on ‘wind-dispersing and exterior syndrome-relieving’ may be the 16 compounds including dihydrodylate glycoside, emodin, rhein, phillyrin, forsythoside E, saikosaponin d, polydatin, verbascoside, etc, which provides a basis for the determination of quality markers.

Spectrum-effect relationship of anti-inflammatory activity of Shufeng Jiedu Capsule based on neural network analysis / 中草药

Objective: To establish the spectrum-effect relationship network model of Shufeng Jiedu Capsule (SJC) for anti- inflammatory activity and explore potential effective compounds of its anti-inflammatory effects. Methods: On the basis of the establishment of fingerprint analysis method for SJC, the uniform design method was used to divide eight herbs in SJC into groups of different proportions to determine their inhibition ratios of TNF-α and IL-6 released by LPS-induced RAW264.7 cells. Pharmacodynamic data and chemical information of MS fingerprints of each group were analyzed with the BP artificial neural network to get the anti-inflammatory effect of each peak to establish the chromatography-efficacy relationship. Results: According to the optimal neural network model, 14 characteristic peaks were calculated and found to be significantly correlated with the anti- inflammatory activity of SJC. Conclusion: Through the spectrum-effect study, it is speculated that the anti-inflammatory components of SJC may be the 14 compounds including rhein, emodin, verbenalin, verbascoside, pinoresinol-β-D-glucoside, forsythoside A, polydatin, etc., which provides a reference for the quality control and determination of effective components.

Approaches and methods of property-flavour material basis of Chinese materia medica based on molecular docking technology of taste and olfactory receptors / 中草药

The theory on the property of Chinese materia medica (CMM) is one of the core contents of CMM theory, among which five flavors theory is the core content of CMM theory. The objective characterization of the five flavors of CMM is the key to the theoretical study of CMM, which is an important way to explain the scientific connotation of CMM. Based on the basic definition of five tastes medicinal properties and the interaction of drug molecules-taste, olfactory receptors, authors put forward the research methods using molecular docking technology and the research approach. In this study, the expression of five tastes drug property was systematically discussed from the aspects of biological mechanisms of taste, olfactory perception, molecular docking, homology modeling, and receptor selection, which provides a reference method for the study of drug property theory from the perspective of taste and smell attribute of five tastes.

Identification of chemical constituents of Yuanhu Zhitong prescription by HPLC-QTOF/MS / 药学学报

This study was designed to clarify the chemical constituents in Yuanhu Zhitong prescription (YHZT), a rapid high performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry (HPLC-QTOF/MS) method was established. Based on the high resolution MS spectra data, fragmentation ion information, reference standards data and literature reports, 51 peaks including 28 alkaloid compounds and 23 coumarin compounds were identified. The chemical constituents in YHZT were rapidly, accurately, systematically analyzed. The results lay a foundation for the quality control of effective compounds of YHZT.

Study on compatibility rationality of Yuanhu Zhitong Dropping Pills for treatment of primary dysmenorrhea based on network pharmacology / 中草药

Objective: To explore the compatibility rationality of Yuanhu Zhitong Dropping Pills (YZDP) for the treatment of primary dysmenorrhea. Methods: The potential targets and pathways of tetrahydropalmatine, palmatine, glaucine, protopine, imperatorin, and isoimperatorin, selected as the representative compounds of alkaloid and coumarin from Corydalis Rhizoma and Angelicae Dahuricae Radix, were predicted by the methods of network pharmacology. The results were verified by docking and the mechanism and compatibility regularity of YZDP were analyzed by integrating all the data. Results: The in silico prediction results showed that the six compounds of YZDP affected 19 related pathways through 23 target proteins. The pathways were involved in central analgesia, hormonal regulation, spasmolysis, vasodilatation, inflammation, and immunoregulation. The alkaloid and coumarin showed common targets and pathways but also had their own emphasis. The pathways were connected by the common targets, showing the synergistic effect of different compounds by acting on multi-targets and multi-pathways. Conclusion: Corydalis Rhizoma acts as monarch drug, showing the effect of analgesia, regulating qi, and activating blood by acting on thromboxane receptors, angiotensin receptors, and targets related with central analgesia and spasmolysis. Angelicae Dahuricae Radix, showing the effect of regulating qi, and activating blood by regulating the pathways of spasm, inflammation, and so on, acts as ministerial drug to assist monarch drug to enhance treatment which reflected the compatibility rationality of Corydalis Rhizoma and Angelicae dahuricae Radix.

Chemical onstituents and mechanism of Corydalis Rhizoma based on HPLC-QTOF/MS and G protein-coupled receptor analysis / 药学学报

The chemical constituents of Corydalis Rhizoma were identified in the 60% ethanol extract using high performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry (HPLC-QTOF/MS). The stimulation and inhibition effects of Corydalis Rhizoma and its representative compounds (protopine, palmatine, tetrahydropalmatine) on G protein-coupled receptor (GPCR), including 5-hydroxytryptamine 1A receptor (5-HT1A), μ opioid receptor (OPRM1), β2 adrenergic receptor (ADRB2), dopamine receptor (D2), acetylcholine receptor (M2) and thromboxane-prostaglandin receptor (TP) were explored using the fluorescence assay of intracellular calcium ion. As a result, 31 compounds were obtained and 28 alkaloid compounds were identified. The results of GPCR experiments showed that Corydalis Rhizoma could activate 5-HT1A, OPRM1, ADRB2 receptors and block D2 receptor. Protopine showed antagonism on D2 and M2 receptors, tetrahydropalmatine could agitate ADRB2 receptor and antagonize D2 and TP receptors, while palmatine showed no significant biological activity on the 6 GPCRs. In conclusion, Corydalis Rhizoma may exert biological activity by multi-components acting on multi-targets.

Network pharmacology-based study on mechanism of Yuanhu Zhitong Dropping Pills in the treatment of primary dysmenorrhea / 药学学报

This study was designed to explore the mechanism of Yuanhu Zhitong Dropping Pills (YHZT) in the treatment of primary dysmenorrhea by pharmacological network technology and establish a research approach of "Compound-Target-Pathway-Disease" network. Twenty-eight compounds absorbed into blood including 22 prototype and 6 metabolites of YHZT were submitted to PharmMapper and Kyoto Encyclopedia of Genes and Genomes (KEGG) bioinformatics softwares to predict the target proteins and related pathways respectively. The network of "Compound-Target-Pathway-Disease" was constructed and analyzed using Cytoscape software. The in silico prediction results showed that the 28 constituents of YHZT affected 111 pathways through 109 target proteins. Among them, a total of 52 proteins and 31 pathways were related to the primary dysmenorrhea. The effect of YHZT on primary dysmenorrhea may be dependent on regulation of the proteins and pathways related with hormonal regulation, central analgesia, spasmolysis, inflammation and immunoregulation.

Study on anti-inflammatory components in Rosae Rugosae Flos based on bioactivity-integrated UPLC-Q/TOF method / 中草药

Objective: To investigate the anti-inflammatory effect of Rosae Rugosae Flos (RRF) and screen its bioactive components on nuclear factor-κappa B (NF-κB) inhibition by ultra-performance liquid chromatography coupled with quadrupole/time of flight mass spectrometry (UPLC-Q/TOF MS). Methods: The anti-inflammatory effect of RRF was assessed in Pseudomonas aeruginosa strain-induced acute lung infection mouse model. UPLC-Q/TOF MS coupled with NF-κB activity luciferase reporter assay system was applied to detect the potential anti-inflammatory components in RRF extract. Results: The administration of RRF extract could ameliorate the neutrophil infiltration and diminish the levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interlukin-6 (IL-6), and IL-8 in P. aeruginosa induced mice. Seven components were screened to have potential NF-κB inhibitory effects based on the bioactivity-integrated UPLC-Q/TOF assay system. Conclusion: RRF can contribute to the alleviation of inflammation in mice with acute lung infection induced by P. aeruginosa strain. Seven potentially active ingredients, such as gallic acid, 4-O-galloylquinic acid, methylgallate, centaureidin, strictinin, casuariin, and ellagic acid are found to have anti-inflammatory effects as NF-κB inhibitors.