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OBJECTIVE To analyze the main components of Chelidonii Herba-Corydalis Rhizoma （CHCR）， and to predict pharmacodynamic substances against estrogen receptor （ER） -positive breast cancer and their potential targets and signaling pathways， followed by verifying experiments. METHODS The ethanol extract of CHCR was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The network pharmacology analysis was performed for the screened components. The network diagram of CHCR “active components-target-pathway” was constructed， and the enrichment pathway in vitro was validated. RESULTS A total of 58 chemical components were identified， including 57 alkaloids and 1 organic acid. A total of 38 active ingredients were screened from the network pharmacology， and 38 core targets were found in the protein-protein interaction network of “component-disease” intersection targets； 258 gene ontology entries and 137 Kyoto encyclopedia of genes and genomics pathways were obtained， mainly including estrogen signal pathway， phosphatidylinositol-3-kinase/protein kinase B （PI3K/Akt） signal pathway， etc. The results of validation test showed that the median inhibitory concentration of CHCR to MCF-7 cells was 693 μg/mL； 150， 300， 600 μg/mL CHCR could significantly reduce the expressions of phosphorylated PI3K， phosphorylated Akt， ERα protein and ESR1 mRNA （P＜0.01）. CONCLUSIONS The anti-ER-positive breast cancer effect of CHCR may be related to the regulation of ER and PI3K/Akt pathways， which has the characteristics of multi-component and multi-target effects.
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A chemical investigation on the aqueous extract of Corydalis yanhusuo tubers led to the isolation and structural elucidation of three pairs of trace enantiomeric hetero-dimeric alkaloids, (+)/(-)-yanhusamides A-C ( 1- 3), featuring an unprecedented 3,8-diazatricylco[5.2.2.02,6]undecane-8,10-diene bridged system. Their structures were exhaustively characterized by X-ray diffraction, comprehensive spectroscopic data analysis, and computational methods. Guided by the hypothetical biosynthetic pathway for 1- 3, a gram-scale biomimetic synthesis of (±)- 1 was achieved in 3 steps using photoenolization/Diels-Alder (PEDA) [4+2] cycloaddition. Compounds 1‒3 exhibited potent inhibition of NO production induced by LPS in RAW264.7 macrophages. The in vivo assay showed that oral administration of 30 mg/kg of (±)- 1 attenuated the severity of rat adjuvant-induced arthritis (AIA). Additionally, (±)- 1 induced a dose-dependent antinociceptive effect in the acetic acid-induced mice writhing assay.
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Deciphering the metabolites of multiple components in herbal medicine has far-reaching significance for revealing pharmacodynamic ingredients.However,most chemical components of herbal medicine are secondary metabolites with low content whose in vivo metabolites are close to trace amounts,making it difficult to achieve comprehensive detection and identification.In this paper,an efficient strategy was proposed:herb-derived metabolites were predicted according to the structural characteristics and metabolic reactions of chemical constituents in Corydalis Rhizoma and chemical structure screening tables for metabolites were conducted.The fragmentation patterns were summarized from represen-tative standards combining with specific cleavage behaviors to deduce structures of metabolites.Ion abundance plays an important role in compound identification,and high ion abundance can improve identification accuracy.The types of metabolites in different biological samples were very similar,but their ion abundance might be different.Therefore,for trace metabolites in biological samples,we used the following two methods to process:metabolites of high dose herbal extract were analyzed to char-acterize those of clinical dose herbal extracts in the same biological samples;cross-mapping of different biological samples was applied to identify trace metabolites based on the fact that a metabolite has different ion abundance in different biological samples.Compared with not using this strategy,44 more metabolites of clinical dose herbal extract were detected.This study improved the depth,breadth,and accuracy of current methods for herb-derived metabolites characterization.
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(±)-Bicoryanhunine B (1), a new dimeric benzylisoquinoline alkaloid was isolated from the dried tubers of Corydalis yanhusuo by various chromatographic methods, including silica gel, Sephadex LH-20, reverse phase C18, and semi-preparative HPLC. Its structure was determined by spectroscopic methods, including UV, IR, ESI-MS, HR-ESI-MS and 1D/2D NMR. (±)-Bicoryanhunine B (1) was a moderate PD-1/PD-L1 interaction inhibitor with an IC50 value of 7.80 ± 0.49 μmol·L-1. In addition, 1 exhibited potent inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages with an IC50 value of 4.83 ± 2.21 μmol·L-1.
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Bidehydrocorydaline A (1), a new dimeric alkaloid of proberberine connected by a methylenedioxy group, was isolated from the vinegar-prepared Corydalis yanhusuo by various chromatographic methods, including column chromatography over macroporous adsorption resin and silica gel, reverse-phase MPLC, and semi-preparative HPLC. Its structure was determined by spectroscopic methods, including UV, IR, HR-ESI-MS, 1D and 2D NMR. Bidehydrocorydaline A (1) showed potent inhibitory activity against LPS-induced NO production in RAW 264.7 macrophages, with an IC50 value of 2.33 ± 0.57 μmol·L-1.
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Objective To investigate the pharmacological components of "Szechwan Chinaberry Fruit-Rhizoma Corydalis" drug combination and its potential molecular mechanism in the treatment of liver cancer based on network pharmacology. Methods Related databases, such as TCMSP, Uniprot, and GeneCard, were used to obtain the effective components of Szechwan Chinaberry Fruit and Rhizoma Corydalis, their corresponding action targets, and the disease targets of liver cancer, and the intersecting targets of drugs and diseases were selected. In addition, STRING and Metascape databases were used to screen out the core targets of drug action and perform GO function and KEGG pathway enrichment analyses. Results There were 9 active components in Szechwan Chinaberry Fruit and 49 active components in Rhizoma Corydalis, with 1 common component between the two drugs; there were 181 action targets of Szechwan Chinaberry Fruit and 1097 action targets of Rhizoma Corydalis, with 143 common targets between the two drugs. There were 162 intersecting targets between the drug combination and liver cancer, and the main genes involved were IL6, TP53, VEGFA, TNF, and CASP3. KEGG analysis showed that the main pathways involved included cancer pathway, AGE-RAGE signaling pathway of diabetes complications, TNF signaling pathway, NF-κB signaling pathway, and thyroid hormone signaling pathway. Conclusion There are many different components in the drug combination of "Szechwan Chinaberry Fruit-Rhizoma Corydalis", which can exert a therapeutic effect on liver cancer by acting on related genes and signaling pathways.
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OBJECTIVE：To establish the fingerprint of crude/vinegar-processed Corydalis yanhusuo decoction pieces and their dispensing granules，and to determine the contents of five alkaloids （protopine，tetrahydropalmatine，corydaline，berberine hydrochloride，palmatine hydrochloride ）. METHODS ：HPLC method was adopted. The determination was performed on Agilent TC-C18 column with mobile phase consisted of 0.1％ phosphoric acid solution-methanol （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm，and the column temperature was 30 ℃. The sample size was 10 μL. Using palmatine hydrochloride as reference ， Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition） was used to establish the fingerprint of 11 059） batches of C. yanhusuo decoction pieces ，7 batches of crude . yanhusuo dispensing granules ， 12 batches of vinegar- processed C. yanhusuo decoction pieces and 11 batches of vinegar-processed C. yanhusuo dispensing granules. The same HPLC method was adopted to determine the contents of protopine， tetrahydropalmatine， corydaline， berberine hydrochloride and palmatine hydrochloride in 41 batches of crude/ vinegar-processed C. yanhusuo decoction pieces and their dispensing granules. RESULTS ：There were 12 and 20 common peaks for crude C. yanhusuo decoction pieces and its dispensing granules ，and 14 and 16 common peaks for vinegar-processed C. yanhusuo decoction pieces and its dispensing granules. The similarity of each batch of same type were 0.529-0.981，0.342-0.985， 0.711-0.999，0.437-0.998，respectively. The linear range of protopine ，tetrahydropalmatine，corydaline，berberine hydrochloride and palmatine hydrochloride were 1.9-38.0，2.0-40.0，2.2-44.0，2.6-52.0，2.3-46.0 μg/mL（R2＞0.999 0）. The recoveries were 100.12％-100.98％（RSD＝1.05％-1.90％，n＝9）. RSDs of precision ，reproducibility，stability（24 h）and durability tests were all lower than 2.0％. The average contents of five alkaloids in different batches of crude/vinegar-processed C. yanhusuo decoction pieces and its dispensing granules were 0.24-0.46，0.37-0.82，0.24-0.58，0.07-0.75，0.24-0.76 mg/g. RSDs were 12.27％-147.48％. CONCLUSIONS：The fingerprint of crude/vinegar-processed C. yanhusuo decoction pieces and its dispensing granules is established successfully. The similarities of fingerprint are different before and after processing with vinegar ，and the contents of five alkaloids in C. yanhusuo decoction pieces and its dispensing granules are greatly different.
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Objective: To analyze the diversity and community structure of endophytic fungi in Corydali yanhusuo tuber and their correlations with tetrahydropalmatine content. Methods The endophytic fungi in C. yanhusuo tuber from eight localities (Pan’an of Zhejiang Province, Dongyang of Zhejiang Province, Jinyun of Zhejiang Province, Sanhe of Shanxi Province, Dongjiaying of Shanxi Province, Kaixian of Chongqing, Xuancheng of Anhui Province and Xinyang of Henan Province) were isolated and identified according to the morphology and ITS sequences, so as to analyze the community structure of endophytic fungi; HPLC was used to determine tetrahydropalmatine content, and Excel and SPSS were used to analyze their correlations and establish multiple linear regression equations at different levels of dominant genera and species. Results: A total of 1 742 endophytic fungi were isolated and divided into 19 taxa (14 were identified to species level and five to genera level), belonging to 3 phyla, 5 classes, 10 orders, 14 families and 15 genera; The tetrahydropalmatine content in C. yanhusuo tuber from eight localities was higher than the standard of Chinese Pharmacopoeia (2015 edition); The richness (S) and diversity index (H’) of endophytic fungi in C. yanhusuo tuber and tetrahydropalmatine content in Zhejiang Province were extremely significant or significantly higher than those in the other localities. The diversity index (H’) was significantly positive correlated with the tetrahydropalmatine content; There was the largest positive correlation between the tetrahydropalmatine content and T83 (Trichoderma sp.) (correlation coefficient: 0.793). Conclusion: There are abundant endophytic fungi resources in C. yanhusuo; T83 as a dominant endophytic fungus probably related to the accumulation of tetrahydropalmatine.
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To establish a DNA molecular markers method for identification of Corydalis yanhusuo,C. turtschaninovii and C. decumbens,the mat K,trn G and psb A-trn H sequences of 56 samples from 14 species of C. yanhusuo,C. turtschaninovii,C. decumbens and their related species were obtained by sequencing. The SNP loci were obtained by Bio Edit 7. 2. 2 software. The primers for AS-PCR identification were designed based on the mutation sites,and the conditions of PCR were optimized to identify C. yanhusuo,C. turtschaninovii,and C. decumbens according to the specific bands. The results showed that the amount of template( 0. 6-1 200 ng)and annealing temperature( 42-60 ℃) had little influence on the amplification results,and the number of cycles had much influence on the amplification results. When the number of cycles was 20,the specific bands of 297 bp( mat K),353 bp( trn G) and 544 bp( mat K) were amplified from C. yanhusuo,C. turtschaninovii and C. decumbens,respectively. The method established in this study had a minimum detection limit of 6 ng for C. yanhusuo,60 ng for C. decumbens and less than 0. 6 ng for C. turtschaninovii. Thus,the allelespecific PCR method established in the research can specifically identify C. yanhusuo,C. turtschaninovii,and C. decumbens.
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Alleles , Corydalis , Classification , Genetics , Genes, Plant , Genetic Markers , Polymerase Chain ReactionABSTRACT
Objective: To establish a spectrum-effect relationship betweent anti-inflammation effects and extracts of Corydalis yanhusuo, in order to provide ideas and methods for study of material basis of efficacy. Methods UPLC-Q-TOF/MS was used to establishe fingerprints of different extracts of C. yanhusuo, and the flurescent enzyme was used as a marker to perform the anti-inflammation activity test. Finally, the relationships between characteristic peaks and anti-inflammation activity was established by partial least squares regression analysis (PLSR) and gray relational analysis (GRA). The anti-inflammatory component obtained by spectral effect analysis was predicted by molecular docking technology, and its anti-inflammatory mechanism was preliminarily studied. Results:The 95% ethanol extract had significant anti-inflammatory activity. The characteristic peaks of No. 5 and 8-11 were significantly affected in PLSR and GRA. Molecular docking results showed that C. yanhusuo exerted anti-inflammatory effects by acting on PKC, ERK2, IKKβ, JAK1, PI3K-α, PI3K-γ, TNF-α, affecting the transmission of inflammatory signals. Conclusion: The anti-inflammatory effect of C. yanhusuo is the result of the combination of various components. The main anti-inflammatory components are coptisine, berberine, palmatine, dihydrogenine, and dehydrocryptine, which exert anti-inflammatory effects by affecting PI3K, JAK, PKC, ERK, IKKβ, and TNF-α signaling pathways.
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Objective: To study the effect of applying diammonium phosphate on the yield and quality of Corydalis yanhusuo. Methods: By designing different gradient diammonium phosphate amount, the main growth indicators at different fertilizer amount were observed and recorded under the same planting condition. The contents of total ash, alcohol soluble extract, and tetrahydropalmatine were measured and analyzed after harvest according to the requirement of 2015 edition Pharmacopeia. Results: The tubers growth of C. yanhusuo was the most flourishing with the highest drying rate of 33.7% at diammonium phosphate application amount of 375 kg/hm2. Tuber diameter was the largest (2.57 cm), the fresh yield was the highest (5 482.2 kg/hm2), and the yield was increased to 38.2% than that without fertilizer cultivation. The content of total ash (3.0%) was the lowest. The contents of alcohol soluble extract and tetrahydropalmatine were 24.8% and 0.128%, respectively. Conclusion: Applying diammonium phosphate could increase the yield and quality of C. yanhusuo.
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Objective:To optimize the extraction technology of tetrahydropalmatine in Corydalis yanhusuo. Methods:An L9 (34 ) orthogonal experiment was performed to investigate the effects of ethanol concentration, ethanol amount, extraction temperature and ex-traction time on the extraction results with the content of tetrahydropalmatine as the evaluation index. Results:The optimum conditions were as follows:the solid-liquid ratio was 1 :10 ( g·ml-1 ) , 60% ethanol was used as the extraction solvent, the extraction tempera-ture was at 80 ° C for three-time extraction, and the extraction duration for each time was 1 h. Conclusion: The optimized extraction technology for tetrahydropalmatine from Corydalis yanhusuo is stable, reasonable, feasible and easy-operated with low energy consump-tion, which can enhance the comprehensive utilization of Corydalis yanhusuo.
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Corydalis yanhusuo and its active ingredients dl-THP have clear efficacy of analgesia, as well as the effect on vasodilatation, cardiac muscle protection, antibiosis, and anti-inflammation, thus they are used for the treatment of various disease. In recent years, C. yanhusuo is widely used for the treatment of drug addiction. In this paper, the research progress in the chemical constituents and pharmacologic actions of C. yanhusuo are reviewed, which provide the scientific basis for its modernization research and comprehensive utilization.
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Objective: To assess the relative contributions of postharvest processing and geographical source to phytochemical variation of Corydalis Rhizoma, and rhizome of Corydalis yanhusuo, and to examine what phytochemical components are the most sensitive to the differences of each factor and how they change. Methods: HPLC fingerprinting and LC-MS coupled with chemometric approaches were applied. Results: The results of principal component analysis (PCA) and hierarchical cluster analysis (HCA) explicitly demonstrated the postharvest processing could produce a greater impact on the phytochemical profiles of Corydalis Rhizoma than geographical source. The contents of most compounds increased after water boiling while decreased after sulphur-fumigation. Protopine, coptisine, and palmatine were the most variable components in processing. Geographical sources also led to a remarkable phytochemical differentiation, in which the environmental variation of the three regions might play a role. Dehydrocorybulbine, coptisine, dehydrocorydaline, and protopine varied most among the three production regions and decreased sequentially in Zhejiang, Shaanxi, and Jiangsu provinces, China. Conclusion: Both postharvest processing and geographical source should be enhanced with the priority for the former in the quality control of Corydalis Rhizoma. The application of boiling is supported but the consistency should be improved in practice. Sulphur-fumigation is strongly suggested to be abandoned. © 2013 Tianjin Press of Chinese Herbal Medicines.
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OBJECTIVE: To predict the anti-diabetes effects of Corydalis yanhusuo alkaloids with pharmacological network technology and verify the results in animal models. METHODS: Targets of Corydalis yanhusuo alkaloids were collected from Pubmed, CNKI, Wangfang and VIP databases, and targets for treatment of diabetes and diabetic complications were searched from OMIM database. The "components-targets-diabetes" network was constructed and analyzed with cytoscape 2.8.2. Diabetes model was established by STZ injection. RESULTS: The targets of Corydalis yanhusuo alkaloids covered many kinds of protein, including ion channel, G coupled protein, and signal proteins such as potassium channel, MAPK/ERK, VEGF and NOS3. In the network, Corydalis yanhusuo alkaloids associated with diabetes and diabetic complications, especially, non-insulin dependent diabetes, obesity, and diabetic complications via modulation of NOS3, VEGF, INSR, KCNJ11 and IRS1, indicating potential effects of Corydalis yanhusuo for diabetes and diabetic complications. Corydalis yanhusuo alkaloids decreased blood glucose in normal and diabetic ICR mice and improved sugar tolerance in insulin-resistant mice, which was in conformity with the prediction. CONCLUSION: Corydalis yanhusuo alkaloids show potential effects on diabetes and diabetic complications.
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OBJECTIVE:To investigate the dissolution of Corydalis yanhusuo fine powder and its ultrafine powder in order to provide theory basis for improving the bioavailability of preparation.METHODS:The paddle method was applied to prepare test sample solution and HPLC was used to determine the content of tetrahydropalmatine.The dissolution of C.yanhusuo fine powder and its ultrafine powder were calculated.RESULTS:The dissolution and dissolution rate of C.yanhusuo ultrafine powder were all higher than its fine powder.CONCLUSION:Micronization contributes to the dissolution of effective components.
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Objective To find out the optimal supercritical CO2 fluid extr ac tion prosess for Tetrahydropalmatine. Methods The extraction process was optimiz ed by orthogonal design with the yield of Tetrahydropalmatine as markers. Three factors including extraction pressure, temperature and the volume of entrainer were observed. Results The optimal condition of supercritical CO2 fluid extrac tion process was: under the pressure of 15Mpa at 40 ℃ , 1.5 fold of 95 % EtO H with Corydalis yanhusuo W.T.Wang as the entrainer. Conclusion The optimal supe rcritical process is feasible, with the advantage of low temperature and energy consumption, short time, high production and no organic residua.