Effect of Compatibility of Wujiwan on Pharmacokinetics and Tissue Distribution of Representative Components / 中国实验方剂学杂志

ObjectiveTo study the plasma pharmacokinetics and tissue distribution of five representative components in Wujiwan， and to illustrate the difference of metabolism and tissue distribution before and after compatibility. MethodHealthy male SD rats were divided into four groups， including Wujiwan group（A group， 62.96 g·L-1）， Coptidis Rhizoma group（B group， 38.4 g·L-1）， processed Euodiae Fructus group（C group， 5.88 g·L-1） and fried Paeoniae Radix Alba group（D group， 18.68 g·L-1）， with 65 rats in each group， and were administered the drugs according to the clinical dose of decoction pieces converted into the dose of the extracts. Then plasma， liver， small intestine and brain were taken at pharmacokinetic set time in each group after administration. Ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry was developed for the quantitative analysis of five representative components［berberine（Ber）， palmatine（Pal）， evodiamine（Evo）， rutecarpine（Rut） and paeoniflorin（Pae）］ in Wujiwan， their concentrations in plasma， liver， small intestine and brain were detected at different time， plasma samples were processed by protein precipitation， and tissue samples were pretreated by protein precipitation plus liquid-liquid extraction. Non-atrioventricular model was used to calculate the pharmacokinetic parameters of each component， and the parameters of each group were compared. ResultPharmacokinetic results of A group showed that area under the curve（AUC0-t） of the five representative components were ranked as follows：Ber and Pal were small intestine>liver>blood， Evo and Rut were liver>small intestine>plasma， Pae was small intestine>plasma， which was not detected in the liver， no other components were detected in brain except for Ber. In comparison with plasma and other tissues， peak concentration（Cmax） of Ber， Pal， Evo， and Rut were the highest and time to peak（tmax） were the lowest in the liver of A group. In plasma， the AUC0-t and Cmax of Evo and Rut were increased in A group compared with C group， tmax of Pea was elevated and its Cmax was decreased in A group compared with D group. In the liver， compared with B-D groups， Cmax values of 5 representative components except Pae were elevated， AUC0-t of Pae was decreased and AUC0-t of Evo and Rut were increased in the A group. In the small intestine， half-life（t1/2） of each representative components in A group was elevated and tmax was decreased， and Cmax of each representative ingredient except Pal was decreased， AUC0-t values of Ber and Pal were increased， whereas the AUC0-t values of Evo and Rut were decreased. ConclusionThe small intestine， as the effector organ， is the most distributed， followed by the liver. The pharmacokinetic parameters of the representative components in Wujiwan are changed before and after compatibility， which is more favorable to the exertion of its pharmacodynamic effects.

Comparison of Effect of Euodiae Fructus Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Oxidative Stress and Autophagy in Mouse Liver Under Toxic Dose / 中国实验方剂学杂志

ObjectiveTo investigate the principle and scientific connotation of Euodiae Fructus（EF） processed with Glycyrrhizae Radix et Rhizoma（Gly） by comparing the effects of unprocessed products of EF（UEF） and processed products of EF with the different proportions of Gly（GEFs） at toxic doses on oxidative stress and autophagy in the liver of mice. MethodSeventy mice were randomly divided into 7 groups， namely the control group， the UEF group， the group of the processed products of EF without Gly（PEF） and 4 groups of GEFs（the mass ratios of EF to Gly were 100∶3， 100∶6， 100∶12 and 100∶24， respectively， hereinafter referred to as the processed products of EF with the mass ratios of 100∶3， 100∶6， 100∶12 and 100∶24 of Gly）. The mice were given purified water， the decoction of UEF， PEF and GEFs by gavage at a dose of 30 g·kg-1. PEF and GEFs were prepared according to the method under EF in the 2020 edition of Chinese Pharmacopoeia. Levels of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were determined by ultraviolet-visible spectrophotometry， hematoxylin-eosin（HE） staining was used to evaluate the pathological changes of liver tissue， the level of reactive oxygen species（ROS） was detected by fluorescence method， the mRNA expression of heme oxygenase-1（HO-1）， quinone oxidoreductase-1（NQO1）， glutathione-S-transferase 1（GSTA1）， Kelch-like epichlorohydrin-associated protein 1（Keap1） and p62 were measured by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， western blot was used to detect the protein expression of phosphorylated mammal target of rapamycin（p-mTOR）， phosphorylated ribosomal p70 S6 protein kinase（p-p70S6K）， p62， microtubule-associated protein 1 light chain 3Ⅰ（LC3Ⅰ） and LC3Ⅱ. ResultCompared with the control group， after 7 d of administration， the increase in body mass of mice in the UEF group began to slow down and the difference gradually increased， and the liver body index significantly increased（P<0.01）， pathomorphological observation showed that the structure of hepatic lobules was disordered， and local hepatic sinuses were narrowed or disappeared， and there were inflammatory infiltration and local bleeding， the levels of ALT and AST in serum and ROS in liver tissue were significantly increased（P<0.01）， and the expressions of Keap1， HO-1， NQO1， GSTA1， p62 mRNA and p-mTOR， p-p70S6K， p62 protein in liver tissue were significantly decreased（P<0.01）， and LC3Ⅱ/LC3Ⅰ was significantly increased（P<0.01）. Compared with the UEF group， the body mass of mice increased， and the liver body index， the levels of ALT and AST in serum， and the level of ROS in liver tissue all decreased in the groups of PEF and GEFs. Among these groups， only the liver lobules in GEF（100∶6） group were intact， and the size of liver sinuses was close to that in the control group. The mRNA expressions of Keap1， HO-1， NQO1， GSTA1 and p62 in liver tissue showed an overall upward trend in the groups of PEF and GEFs. Among these groups， only the ones of the above mRNA in the GEF（100∶6） group had a significant increase（P<0.05， P<0.01）. The protein expressions of p-mTOR， p-p70S6K， p62 and LC3Ⅱ/LC3Ⅰ had a callback in the groups of PEF and GEFs， of which the protein expressions of p-mTOR， p-p70S6K and LC3Ⅱ/LC3Ⅰ in the GEF（100∶6） group and the expression of p62 protein in the GEF（100∶24） group had the largest callback. Except for p-mTOR protein， other protein expressions were statistically significant（P<0.05， P<0.01）. ConclusionThe hepatotoxicity of EF is closely related to its ability to induce oxidative stress， which leads to pathological autophagy and hepatocyte damage. This ability can be reduced by the processing with different proportions of Gly， especially the ratio of 100∶6.

Hot water washing processing technology of Euodiae Fructus based on change laws of active components and tastes / 中国中药杂志

In order to establish the standardized processing technology of the hot water washing of Euodiae Fructus, this study, based on the traditional processing method of hot water washing of Euodiae Fructus recorded in ancient works and modern processing specifications of traditional Chinese medicine decoction pieces, took the yield of decoction pieces and the content of main components as the indicators and optimized the processing conditions by orthogonal test based on the results of single factor investigation. At the same time, electronic tongue technology was used to analyze the change law of the taste index of Euodiae Fructus during the hot water washing. The results of the single factor investigation showed that the content of the main components in Euodiae Fructus showed some regular changes during the processing. Specifically, the content of chlorogenic acid, hyperin, isorhamnetin-3-O-rutinoside, isorhamnetin-3-O-galactoside, and dehydroevodiamine decreased significantly, with average decreases of-23.75%,-27.80%,-14.04%,-14.03%, and-13.11%, respectively. The content of limonin increased significantly with an average increase of 19.83%. The content of evodiamine, rutaecarpine, evocarpine, and dihydroevocarpine showed fluctuating changes and generally increased, with average variation amplitudes of 0.54%,-3.78%, 2.69%, and 5.13%, respectively. The orthogonal test results showed that the optimum processing parameters for the hot water washing of Euodiae Fructus were as follows: washing time of 2 min, the solid-to-liquid ratio of 1∶10 g·mL~(-1), washing temperature of 80 ℃, washing once, and drying at 50 ℃. After the hot water washing processing, the average yield of Euodiae Fructus pieces was 94.80%. The content of limonin, evodiamine, and rutaecarpine was higher than those of raw pro-ducts, and the average transfer rates were 102.56%, 103.15%, and 105.16%, respectively. The content of dehydroevodiamine was lower than that of the raw products, and the average transfer rate was 83.04%. The results of taste analysis showed that the hot water washing could significantly reduce the salty, astringent, and bitter tastes of Euodiae Fructus. This study revealed the influence of the hot water washing on the content of main components and taste of Euodiae Fructus, and the processing technology of the hot water was-hing of Euodiae Fructus established in this study was stable, feasible, and suitable for industrial production, which laid a foundation for clarifying its processing principle and improving the quality standard and clinical application value of decoction pieces.

Explanation of Scientific Connotation of Euodiae Fructus Stir-fried with Coptidis Rhizoma Based on UPLC-Q-TOF/MS and Pattern Recognition Technology / 中国实验方剂学杂志

ObjectiveTo analyze changes of the chemical composition in Euodiae Fructus before and after processing with Coptidis Rhizoma decoction， so as to provide scientific basis for elucidating the processing mechanism of this decoction pieces. MethodUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MS） was performed on a Titank C18 column （2.1 mm×100 mm， 1.8 μm）， the mobile phase was 0.1% formic acid aqueous solution-acetonitrile for gradient elution， the column temperature was set at 40 ℃， the flow rate was 0.25 mL·min-1. Electrospray ionization （ESI） was used to scan in positive and negative ion modes， and the scanning range was m/z 50-1 250. The chemical constituents in Euodiae Fructus were identified before and after processing by reference substance comparison， database matching and literature reference， and MarkerView™ 1.2.1 software was used to normalize the obtained data， SIMCA-P 14.1 software was employed to perform principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） on MS data of raw and processed products to screen the differential components before and after processing. ResultA total of 50 compounds were identified， including 48 kinds of stir-fried products with Coptidis Rhizoma decoction and 44 kinds of raw products. After processing， six compounds were added， including danshensu， noroxyhydrastinine， oxyberberine， 13-methylberberrubine， protopine and canadine. However， two kinds of compounds， including （S）-7-hydroxysecorutaecarpine and wuchuyuamide Ⅱ， were not detected after processing. In general， after processing， the overall contents of phenolic acids and flavonoids decreased significantly， the overall content of limonoids increased， and the overall content of alkaloids did not decrease insignificantly. The results of PCA and OPLS-DA showed that there were significant differences in the composition and content of the chemical components of Euodiae Fructus before and after processing， and a total of 12 variables such as quercetin， dihydrorutaecarpine and dehydroevodiamine were obtained by screening. ConclusionEuodiae Fructus stir-fried with Coptidis Rhizoma decoction mainly contains phenolic acids， flavonoids， limonoids and alkaloids. The composition and content of the chemical components have some changes before and after processing. The addition of processing excipients and hot water immersion are the main reasons for the difference， which can provide experimental basis for interpretation of the processing mechanism of this characteristic processed products of Euodiae Fructus.

Analysis of Zuojinwan Based on Ancient Literature / 中国实验方剂学杂志

Zuojinwan originated from Danxi′s Experiential Therapy （《丹溪心法》） in the Yuan dynasty. It is a representative prescription for the treatment of liver fire invading stomach syndrome， and is also one of the typical prescriptions of the anti-adjuvant method of traditional Chinese medicine （TCM）. In this paper， the method of bibliometrics was used to systematically sort out the ancient books of Zuojinwan， and 729 relevant literature data were obtained. After certain retrieval and screening， 57 ancient books of TCM were finally obtained. The statistics and analysis were carried out from the aspects of prescription source， historical evolution， composition， functions， evolution of prescription meaning， prescription dose， and preparation and usage of Zuojinwan. It was found that Zuojinwan was composed of Coptis chinensis rhizoma and Euodia rutaecarpa fructus in a ratio of 6∶1. It was mainly used for the treatment of liver fire invading stomach syndrome. The symptoms included pain in chest and hypochondrium， vomiting and bitter mouth， noisy acid-swallowing， red tongue coating yellow， and pulse string number. Later medical records recorded that Zuojinwan was mostly consistent with the original prescription. It mainly treated various diseases caused by liver fire， including left by liver fire， including left hypochondriac pain， swallowing acid and vomiting acid， tendon hernia and lump， epigastric pain， bitter mouth pulse string， head pain， diarrhea， gonorrhea， cold and hot， abdominal pain， alcohol wet yellowing， silence of oral dysentery and so on. There was little controversy in the analysis of relevant prescriptions. In the past dynasties， pills was mainly used， which was consistent with the original prescription. In modern times， it is mainly water flooding for pills or steamed cakes for pills， warm boiling water to serve 6 g， taking 2-3 g per time， the history is basically the same. In this paper， through the excavation， collation and systematic analysis of the ancient literature of TCM that recorded Zuojinwan， we hope to provide the literature basis for the development， inheritance and utilization of this famous classical formulas.

Research Progress on Compatibility Mechanism of Coptidis Rhizoma-Euodiae Fructus： A Review / 中国实验方剂学杂志

Herb pair， a common form of compounding in Chinese medicinal prescriptions， reflects the experience of pharmacists in clinical medication in the past. It is characterized by simple composition while has the basic characteristics of Chinese medicine compounding. The combination of two medicinal herbs can enhance effect or reduce toxicity. Coptidis Rhizoma （CR） has the effects of clearing heat， drying dampness， purging fire， and removing toxin. Euodiae Fructus （EF） is acrid， bitter， and hot-natured， which can not only warm the Yang Qi in spleen and stomach to dissipate cold and relieve pain， but also descend stomach Qi and prevent vomiting. Furthermore， it can warm the liver and kidney. CR and EF form a typical cold-heat herb pair， which oppose and yet also complement each other. Specifically， their cold and heat natures interact with each other to clear liver fire， harmonize stomach for descending adverse Qi， relieve depression， and dissipate mass. CR clears the intestine and stops dysentery， while EF warms the middle and promotes the circulation of Qi. The combination of them can thus clear heat， dry dampness， and relieve pain. Modern pharmacological studies have demonstrated that CR-EF has not only significant efficacy against digestive system diseases but also good anti-cancer， anti-inflammatory， anti-ulcer， and lipid-lowering activities. Therefore， the article summarized the effect enhancement and toxicity reduction of the herb pair at the levels of cellular molecule， isolated organ， and whole animal， and clarified the mechanism of its pharmacological action. It will provide a theoretical basis for further development and clinical use of the herb pair.

Study on effect of gastrointestinal function and portal vein hemodynamics with abdominal hot compressing with evodiae fructus and crude salt after hepatectomy / 国际中医中药杂志

Objective:To study on the effects of gastrointestinal function and portal vein hemodynamics applicated with abdominal hot compressing with evodiae fructus and crude salt after hepatectomy.Methods:A total of 60 patients who underwent hepatectomy were randomly divided into 2 groups by random number table method, with 30 in each group. The control group was treated with conventional basic western medicine, while the treatment group was treated with medicinal evodiae fructus and crude salt hot compress on abdomen on the basis of the control group. The portal vein diameter (PVD), portal venous flow velocity (PVV), recovery time of main clinical indexes, clinical symptom scores and liver function indexes were dynamically monitored at different observation time points.Results:The scores of abdominal distension, nausea and vomiting in the treatment group were significantly lower than those in the control group 3 days after treatment ( t values were -3.489 and -2.740, respectively, all Ps<0.05). The recovery time of bowel sounds, first exhaust time and first defecation time in the treatment group were significantly earlier than those in the control group ( t values were -3.622, -4.297 and -4.151, respectively, all Ps<0.01). With the extension of treatment time, ALT in 2 groups showed a gradual downward trend ( P<0.05 or P<0.01), DBIL in control group was significantly higher at 3 days after treatment than before ( t=-2.157, P=0.039), and TBIL was significantly lower at 7 days after treatment than before ( t=2.175, P=0.038). The PVD ( t values were 3.528, 2.160) and PVV ( t values were 11.096, 4.264) of the treatment group were significantly higher than those of the control group 3 and 5 days after treatment ( P<0.01 or P<0.05). Conclusion:Early application of abdominal hot compressing with evodiae fructus and crude salt hot compress on abdomen after hepatectomy can improve the portal vein blood circulation and promote the rehabilitation of gastrointestinal function in patients with hepatectomy.

Analysis of Chemical Constituents in Euodiae Fructus by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To analyze the chemical constituents in Euodiae Fructus by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. Method:The chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with acetonitrile （A）-0.1% formic acid aqueous solution （B） as mobile phase （0-3 min， 6%A； 3-4 min， 6%-10%A； 4-7 min， 10%-12%A； 7-8 min， 12%-14%A； 8-13 min， 14%-15%A； 13-15 min， 15%-20%A； 15-18 min， 20%-30%A； 18-21 min， 30%-49%A； 21-25 min， 49%-51%A； 25-27 min， 51%-73%A； 27-30 min， 73%-80%A； 30-31 min， 80%-100%A； 31-32 min， 100%A） for gradient elution. The column temperature was 35 ℃， and the flow rate was 0.4 mL·min^-1. Mass spectrometry was performed using an electrospray ionization and data were collected in positive and negative ion modes， and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 200. The chemical constituents in Euodiae Fructus were identified rapidly and comprehensively based on the accurate relative molecular mass and combined with literature data and reference substances. Result:A total of 92 chemical constituents were speculatively identified from the 70% methanol extract of Euodiae Fructus， including 39 alkaloids， 19 flavonoids， 12 limonoids， 20 phenolic acids and 2 organic acids. Among them， 26 compounds were confirmed by the reference substances. Conclusion:The compound types of Euodiae Fructus are multiple and quite different in polarity. The chemical compositions of Euodiae Fructus from different regions and species are similar. The established method is rapid and accurate， with which the chemical compositions of Euodiae Fructus have been identified comprehensively. Therefore， this study provides an experimental reference for further clarifying active and toxic constituents of Euodiae Fructus.

Investigation of Transdermal Constituents and Molecular Mechanism of Euodiae Fructus in Treatment of Diarrhea by Transdermal Drug Delivery Based on Integrated Pharmacology and UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:To identify the transdermal constituents of Euodiae Fructus and predict its molecular mechanism in treating diarrhea by transdermal drug delivery. Method:Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS） and integrated pharmacology methods were used. The rapid identification of transdermal constituents of Euodiae Fructus was realized by the means of comparison of reference substances， analysis of UNIFI system and mass spectrometry. On this basis， Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0， SymMap， DisGeNET databases and literature were used to collected potential targets of transdermal constituents of Euodiae Fructus and targets for diarrhea-related diseases. The disease targets and drug targets were topologically analyzed to obtain the core targets， which were used for the Gene Ontology （GO） function and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. Finally， Cytoscape 3.6.0 was used to build up a network of transdermal constituents-core targets-key pathways. Result:A total of 19 chemical constituents were speculatively identified from Euodiae Fructus extract， including quinolone alkaloids， limonins， indole alkaloids， organic acids and sterols. A total of 174 core targets of Euodiae Fructus for treating diarrhea were obtained by a topology analysis， signaling pathways of inflammatory response， cell proliferation， nutrient regulation and energy metabolism， signal transduction， bacterial infection were obtained through the analysis of KEGG enrichment. Conclusion:In this study， the transdermal constituents of Euodiae Fructus are identified for the first time， they can participate in the regulation of intestinal inflammation， maintain the integrity of intestinal mucosa， repaire and adjust the metabolism of the body by acting on Rac protein family， phosphatidylinositol 3-kinase， cytochrome P450 enzymes and aldo-keto reductase， respectively. In general， the molecular mechanism of Euodiae Fructus in the treatment of diarrhea is preliminarily elucidated.

Potential targets of Euodiae Fructus in treatment of insomnia based on network pharmacology / 中国中药杂志

The acupoint application of Euodiae Fructus at Yongquan(KI1) can significantly improve the sleep quality of patients with insomnia with berberine as the main effective component for the efficacy. Nineteen active compounds and 203 drug targets were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). After comparison with GeneCards and Online Mendelian Inheritance in Man(OMIM), 24 common genes of diseases and drugs were obtained. STRING 11.0 was used to construct a protein-protein interaction(PPI) network of the overlapping genes, and Matthews correlation coefficient(MCC) was employed to screen the core genes, which were then subjected to enrichment analysis with gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG). The results revealed that the main compounds of Euodiae Fructus, such as berberine and rutaecarpine, participated in the biological processes(such as neurotransmitter receptor activity) by regulating C-reactive protein(CRP), estrogen receptor 1(ESR1), 5-hydroxytryptamine(5-HT) receptor, and interleukin-6(IL-6) to exert sedative, anxiolytic, and antidepressant effects. Sixty 4-week-old SPF mice were randomly divided into a control group, a model group, a positive drug(diazepam tablets) group, and low-, medium-, and high-dose berberine groups. Medication with corresponding drugs was performed for one week. The results demonstrated that berberine was potent in reducing the activities and standing times of mice, down-regulating the levels of CRP and IL-6 mRNA in the hypothalamus, and up-regulating the expression of 5-HT(P<0.01); however, no significant effect on ESR1 was observed. The network of Euodiae Fructus in treating insomnia was constructed by network pharmacology and verified by tests. The findings indicated that the therapeutic efficacy of Euodiae Fructus in treating insomnia was achieved by participating in multiple biological processes, such as neurotransmitter receptor activity, which provided a scientific basis for its clinical application.

Network pharmacology-based study on mechanisms of drug pair of Coptidis Rhizoma-Euodiae Fructus in treating cancer and cardio-cerebrovascular diseases / 中草药

Objective: To clarify the characteristics of drug pair of Coptidis Rhizoma-Euodiae Fructus on multi-components, multi-targets and multi-pathways based on the network pharmacology, and provide a theoretical reference for further study on the pharmacodynamic basis and mechanism of Coptidis Rhizoma-Euodiae Fructus against cancer and cardio-cerebrovascular diseases. Methods: Through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database, the parameters of oral bioavailability (OB ≥ 30%) and drug-likeness (DL ≥ 0.18) combined with the biological function, contents, research hotspot and other factors were used to screen out the active ingredients of the medicine pair of Coptidis Rhizoma - Euodiae Fructus. DrugBank and TCMSP databases were used to predict and analyze the targets, and Uniprot database was used to sort out all relevant genes affecting the targets. Combining the KEGG database for GO analysis and pathway analysis, "ingredients-targets-related diseases" network was constructed using Cytoscape software. Results: A total of 41 compounds and 212 potential target genes, as well as 44 signaling pathways were obtained after screening from Coptidis Rhizoma-Euodiae Fructus, and there were 44 major signaling pathways involved in 30 pathways related to cancer, seven pathways which were related to cardio-cerebrovascular diseases, seven pathways which were related to cancer and cardio-cerebrovascular diseases. Conclusion: The active ingredients of Coptidis Rhizoma-Euodiae Fructus can up-regulate the expression of cancer apoptosis genes through multi-components, multi-targets and multi-pathways, down-regulate the expression of cell cycle and apoptosis-related genes, and produce anti-cancer effect; Protecting vascular cells by inhibiting vascular calcification, and to achieve the effect of treating cardio-cerebrovascular diseases by inhibiting apoptosis and proliferation.

Study on hepatotoxicity mechanism of Euodiae Fructus based on network pharmacology / 中草药

Objective: To screen the main hepatotoxic components, predict the target of active components, and explore the mechanism of liver toxicity of Euodiae Fructus (EF). Methods: According to TCMSP database, PubChem database Pharmmapper server and Uniprot KB database information, active constituents of EF were screened, and targets of hepatotoxicity were predicted. The active component-acting target network of EF was constructed by Cytoscape software, while the function and metabolic pathways of target genes were analyzed by KOBAS 3.0 database. Results: Network analysis results demonstrated that 147 potential hepatotoxic components from EF, accompanied with 49 targets like F2, PIM1, MMP13 and MAOB, connecting with cell metabolism, catalytic activity, stimulate the reaction pathways may be associated with EF’s liver toxic effects. Conclusion: Based on network pharmacology methodology, this paper disclosed that many potential toxic components in EF may interact with cell metabolism, catalytic activity and other pathways through multiple targets, leading to produce hepatotoxicity in vivo as a result, which can provide new clues for further researches of the hepatotoxicity mechanism study of EF.

Chemical components and pharmacological action for Euodiae Fructus and predictive analysis on its Q-marker / 中草药

Euodiae Fructus is one of the most commonly used traditional Chinese medicinal materials with rich resource and wide distribution. It has broad prospects for development and utilization. The chemical constituents of Euodiae Fructus are abundant, including alkaloids, bitter principles, volatile oils, flavonoids and so on. Traditionally, the alkaloids and bitter principles are considered as its main pharmacodynamic components. Based on the review of its chemical composition and pharmacological action of herbal medicine, according to the theory of quality marker, the quality maker of Euodiae Fructus were predicted from the aspects of source, chemical composition and traditional efficacy, traditional medicinal properties, new clinical uses, and toxicity, measurable ingredients, blood-entering ingredients and different compatible environments, which provide a basis for defining quality markers and formulating scientific quality standards of Euodiae Fructus.

Molecular Mechanism of Coptidis Rhizoma-Euodiae Fructus for Gastric Cancer Based on Network Pharmacology / 中国实验方剂学杂志

Objective::To identify the main active components of Coptidis Rhizoma and Euodiae Fructus for the treatment of gastric cancer, predict the targets of the common active components in these two herbs, establish the network of active drug components-target genes, and further explore the potential mechanism and effect of Coptidis Rhizoma-Euodiae Fructus for the treatment of gastric cancer. Method::The active components of Coptidis Rhizoma-Euodiae Fructus were screened by Traditional Chinese Medicine Systems Pharmacology Database (TCMSP), gastric cancer targets were predicted and screened by Genecard database platform, " drug-active ingredient-disease-targets" networks were built by Cytoscape (3.7.1) software, and protein interaction networks were built by String database platform. Finally, Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed by using Bioconductor platform and R language. Result::The 14 potential active components of Coptidis Rhizoma and 15 potential active components of Euodiae Fructus were obtained, involving 127 targets related to gastric cancer. There were 33 common targets for Coptidis Rhizoma-Euodiae Fructus-gastric cancer, which played a therapeutic role in gastric cancer mainly by regulating target genes such as PTGS2, PTGS1, AR, RXRA and NOS2, as well as cell apoptosis, p53 signaling pathway and interleukin (IL)-17 signaling pathway. Conclusion::The therapeutic mechanism of Coptidis Rhizoma-Euodiae Fructus reflects the multi-component, multi-target and multi-pathway characteristics of traditional Chinese medicines and provides the scientific basis for further study and the material basis of Coptidis Rhizoma-Euodiae Fructus against gastric cancer.

Effect of Three Chinese Herbs Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Pharmacokinetics of Dapsone in Rats / 中国实验方剂学杂志

Objective::To investigate the processing purpose of Morindae Officinalis Radix (MO), Euodiae Fructus (EF) and Polygalae Radix (PR) processed by Glycyrrhizae Radix et Rhizoma (Gly). Method::The content of dapsone in rat plasma was determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A)-water (B) for gradient elution (0-5 min, 10%-25%A; 5-20 min, 25%A) and detection wavelength was set at 292 nm. PK Solution 2.0 software was used to simulate pharmacokinetic parameters. Result::Within 300 min after dapsone was administrated, compared with the control (CTL) group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of MO (UMO) group. The elimination of dapsone was accelerated and its peak concentration (Cmax) was decreased in the processed products of MO with Gly (GMO) groups, and they had positive correlation with proportion of Gly in GMO. Compared with the CTL group, the elimination of dapsone was slowed down, and its plasma concentration was increased and its peak time (Tmax) was postponed in the unprocessed product of EF (UEF) group, while their Cmax and Tmax were changed in the processed products of EF with Gly (GEF) groups. Compared with the CTL group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of PR (UPR) group, while the elimination was accelerated and its plasma concentration was decreased in the processed products of PR with Gly (GPR) groups. Conclusion::The elimination of dapsone is slowed down in rats administered with UMO, UEF and UPR, while its elimination is accelerated in rats administered with the processed products of these three herbs with different proportions of Gly. Among the proportions, effect of processed products of these three herbs with 100∶6 (ratio of unprocessed product-Gly) on pharmacokinetics of dapsone is not significant.

UPLC fingerprints and multi-component chemical pattern recognition of Euodiae Fructus in different years / 中草药

Objective: To establish fingerprints of Euodiae Fructus in different years by UPLC, analyze them with chemical pattern recognition technology, and determine the content of four quality difference markers, so as to provide new ideas for the research of Euodiae Fructus “Chen Jiuzhiliang” and provide basis for its quality evaluation. Methods: The fingerprints of Euodiae Fructus samples in three different years were established based on UPLC method. The similarity was calculated by the Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine (2012A edition), and the pattern recognition was carried out by principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). Results: UPLC fingerprints of 47 batches of Euodiae Fructus samples (similarity > 0.997) were established, 40 common peaks were identified and 12 of them were identified. PCA and OPLS-DA analysis were used to effectively distinguish three different years of Euodiae Fructus. A total of 22 quality differential markers were screened, and the content of neochlorogenic acid, chlorogenic acid, evodiamine and rutaecarpine were determined. Conclusion: The combination of UPLC fingerprint of Euodiae Fructus and multi-component chemical pattern recognition technology can effectively distinguish Euodiae Fructus in different years and provide reference for “the older, the better” and quality control of Euodiae Fructus.

Confirmation study of hepatotoxic quality marker in Euodiae Fructus decoction / 中草药

Objective: To confirm the hepatotoxic Q-marker of Euodiae Fructus decoction. Methods: The hepatotoxicity of target compounds was studied by hepatotoxicity index and flow cytometry. The targets and target organs were predicted by network pharmacology. The qualitative absorption analysis of 3-O-trans-caffeoylgluconic acid (1), 4-O-trans-caffeoylgluconic acid (2), 2-O-trans-caffeoylgluconic acid (3) was performed by LC-MS. Through the systematic study of Q-marker properties, the confirmation study of hepatotoxicity Q-marker in Euodiae Fructus decoction was performed. Results: Compounds 1, 2, and 3 inhibited the growth of L02 cells in G0/G1 phase and had direct toxicity to L02 cells, which led to hepatotoxicity. The target organ may be the liver by the network pharmacology analysis. After drug administration, compounds 1, 2, and 3 could be absorbed into the blood by prototype. Compounds 1, 2, and 3 had properties of structural confirmation, toxicity confirmation, measurability, and transmissibility. Conclusion: Compounds 1, 2, and 3 could be confirmed as hepatotoxic Q-marker of Euodiae Fructus decoction.

Analysis of major bioactive components and their metabolites in rats' plasma, urine and feces after oral administration of the aqueous extract of Euodiae Fructus by UHPLC-QTOF-MS / 药学学报

An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF/MS)method has been established to analyze the bioactive components of the aqueous extract of Euodiae Fructusand detect the metabolites in rats after oral administration of the aqueous extract. Compounds were detected by analyzing their retention times, high resolution mass data, mass spectra and comparing with the reference substances. As a result, 27 compounds were characterized from the aqueous extract of Euodiae Fructus, and 25 compounds were identified on the basis of the reference substances. 16 parent compounds and 35 metabolites were detected in rats' plasma, urine and feces samples, and all of the parent compounds were determined unambiguously through comparison with the reference substances. Four classes of compounds were identified from the aqueous extract of Euodiae Fructus, including phenolic acids, limonins, flavonoid glycosides and alkaloids. Phenolic acids, flavonoid glycosides and alkaloids were detected in rats' plasma, urine and feces, while limonins were detected in the urine and feces. The main metabolism pathways of these compounds in rats might include hydroxyl, hydrogenation, methylation, sulfating, glucuronidation, and so on. The results of this study provide references in the material basis of Euodiae Fructus in vivo.

Toxic Q-Marker identification based on conventional application of traditional Chinese medicine: Taking Euodiae Fructus as an example / 中草药

Objective: To establish a method for the toxic Q-Marker identification of traditional Chinese medicine (TCM), based on the scientific analysis on the conventional application of TCM and taking Euodiae Fructus as a case study. Methods: Taking the literature research of the conventional application of "Jiu Jian" and "Tang Xi" as the breakthrough point, the components of Euodiae Fructus decoction were characterized by fingerprint and mass spectrometry technology, and the hepatotoxicity of Euodiae Fructus decoction was evaluated by the growth inhibition rate, lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) of normal human liver cells (L02). The "spectrum-toxicity" relationship analysis was performed by grey relational analysis. Results: The hepatotoxicity of Euodiae Fructus decoction first increased and then decreased with the prolongation of boiling time. Through the "spectrum-toxicity" relationship analysis, it was found that the polar components could be the hepatotoxic Q-Marker in Euodiae Fructus decoction, and the polar components were characterized as isomers of caffeoyl gluconic acid. The scientific connotation of "Jiu Jian" and "Tang Xi" was revealed and the hepatotoxic Q-Marker identification method of Euodiae Fructus was established. Conclusion: It is feasible to take the scientific analysis of conventional application as the breakthrough point for the toxic Q-Marker identification of TCM.

Investigation on Resources of Medicinal Plant of Euodiae Fructus / 中国中医药信息杂志

Objective To understand the distribution condition of resources of medicinal plant of Euodiae Fructus;To provide references for resource conservation and standardized plantation of Euodiae Fructus. Methods Literature search, interview survey and field survey were conducted to investigate the resource conditions of the main producing areas of Euodiae Fructus. Samples were collected and TKW of the medicinal plant was weighted. Results Currently, the main producing areas of Euodiae Fructus are provinces of Jiangxi, Guizhou, Hubei, Hunan and part of provinces and places of Guangxi, Anhui, Zhejiang, Shanxi, and Chongqing. Commercial Euodiae Fructus mainly comes from private investors. The wild resources of Euodiae Fructus are few. The TKW of the medicinal plant is within 5.34-45.62 g, with great differences. The national annual output of cultivated Euodiae Fructus is around 3500 t. The production and marketing is in balance. The price for Euodiae Fructus is relatively stable. Conclusion The plantation of Euodiae Fructus is dispersing, with small scale, low degree of standardization and intensification, and great quality differences. Standardized, large-scale, and industrialized plantation base should be established, and the assessment system for medicine quality should be perfected, with a purpose to ensure the qualified and stable production of Euodiae Fructus. Meanwhile, wild resource production should be enhanced to maintain the biological diversity and promote the sustainable development of Euodiae Fructus.