OBJECTIVES: To explore the therapeutic mechanism of Arctium lappa extract for treatment of Post-Viral Pneumonia Pulmonary Fibrosis (PPF). METHODS: The chemical constituents of Arctium lappa extracts were identified using UHPLC-Q-TOF-MS/MS. Mouse models of pulmonary fibrosis established by tracheal instillation of bleomycin were treated with Arctium lappa extract, and body weight changes were recorded and lung tissue pathology was examined using HE and Masson staining. Metabolomics analysis was used to identify the differential metabolites and the associated metabolic pathways in the treated mice. The common targets of viral pneumonia and pulmonary fibrosis were acquired from the publicly available databases, and the core targets and active constituents were screened using the protein-protein interaction (PPI) network, GO and KEGG enrichment analyses, and molecular docking, and a "gene-metabolite" regulatory network was constructed. The expressions of the core targets were detected in the lung tissues of the treated mice using Western blotting. RESULTS: Fifty-three chemical constituents were identified from Arctium lappa extract. In the mouse models of pulmonary fibrosis, treatment with Arctium lappa extract significantly improved weight loss and ameliorated lung inflammation and fibrosis. The differential metabolites in the treated mice were enriched in energy metabolism pathways involving citrate cycle, pentose phosphate pathway, glycolysis, tryptophan metabolism, glutamate metabolism and glutathione metabolism, which regulated the production of energy metabolism intermediates. Twenty-three key active compounds (mostly lignans and phenolic acids) and 82 core targets were screened, which were associated with the non-canonical Smad signaling pathways (including PI3K/AKT, HIF-1, MAPK, and Foxo) that participated in the regulation of energy metabolism. Arctium lappa extract also regulated the expressions of epithelial-mesenchymal transition (EMT)‑related proteins (fibronectin, vimentim, and Snail, etc.) and inhibited MAPK signaling pathway activation. CONCLUSIONS Preliminary findings suggest that Arctium lappa treats fibrosis by regulating metabolism to inhibit EMT and involves the modulation of non-canonical Smad signaling pathways, such as MAPK providing theoretical support for its clinical application and further research in treating PPF.
Arctium/chemistry* ; Animals ; Pulmonary Fibrosis/metabolism* ; Mice ; Metabolomics ; Network Pharmacology ; Plant Extracts/pharmacology* ; Signal Transduction ; Drugs, Chinese Herbal/pharmacology* ; Molecular Docking Simulation

Chemical constituents were isolated and purified from ethyl acetate fraction of Arctium lappa leaves by silica gel, ODS, MCI, and Sephadex LH-20 column chromatography. Their structures were identified with multiple spectroscopical methods including NMR, MS, IR, UV, and X-ray diffraction combined with literature data. Twenty compounds(1-20) were identified and their structures were determined as arctanol(1), citroside A(2), melitensin 15-O-β-D-glucoside(3), 11β,13-dihydroonopordopicrin(4), 11β,13-dihydrosalonitenolide(5), 8α-hydroxy-β-eudesmol(6), syringin(7), dihydrosyringin(8), 3,4,3',4'-tetrahydroxy-δ-truxinate(9),(+)-pinoresinol(10), phillygenin(11), syringaresinol(12), kaeperferol(13), quercetin(14), luteolin(15), hyperin(16), 4,5-O-dicaffeoylquinic acid(17), 1H-indole-3-carboxaldehyde(18), benzyl-β-D-glucopyranoside(19), and N-(2'-phenylethyl) isobutyramide(20). Among them, compound 1 is a new norsesquiterpenoid, and compounds 2-5, 7-8, and 18-20 are isolated from this plant for the first time.
Arctium/chemistry* ; Magnetic Resonance Spectroscopy ; Luteolin/analysis* ; Plant Leaves/chemistry*

Fructus Arctii is a traditional Chinese medicine. The main counterfeit species are the seeds of Arctium tomentosum, Onopordum acanthium, Silybum marianum, Saussurea costus, Amorpha fruticosa. Traditional identification methods or molecular barcoding techniques can identify Fructus Arctii and its counterfeit species. However, the identification of the mixture of it and its spurious species is rarely reported. In this paper, we sequenced the ITS2 sequences of Fructus Arctii and 5 kinds of spurious species mix powder by high-throughput sequencing to identify the mixed powder species and providing new ideas for the identification of Fructus Arctii mix powder. The total DNA in mixed powder was extracted, and the ITS2 sequences in total DNA was amplified. Paired-end sequencing was performed on the DNA fragment of the community using the Illumina MiSeq platform. The sequence was analyzed by the software FLASH, QIIME and GraPhlAn etc. The results showed that the high quality ITS2 sequences of 39910 mix samples were obtained from the mixed samples, of which the total ITS2 sequence of the samples genus was 34 935. Phylogenetic analysis showed that the samples contained Fructus Arctii, A. tomentosum, O. acanthium, S. marianum, S. costus and A. fruticosa. Using ITS2 sequences as DNA barcodes, high-throughput sequencing technology can be used to detect the Fructus Arctii and its spurious specie in mixed powder, which can provide reference for the quality control, safe use of medicinal materials of Fructus Arctii and the identification of mixed powder of traditional Chinese medicine.
Arctium ; chemistry ; classification ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; Drugs, Chinese Herbal ; standards ; Fabaceae ; Fruit ; High-Throughput Nucleotide Sequencing ; Milk Thistle ; Onopordum ; Phylogeny ; Saussurea

The present study was designed to optimize the processing of Fructus Arctii by response surface methodology (RSM). Based on single factor studies, a three-variable, three-level Box-Behnken design (BBD) was used to monitor the effects of independent variables, including processing temperature and time, on the dependent variables. Response surfaces and contour plots of the contents of total lignans, chlorogenic acid, arctiin, and arctigenin were obtained through ultraviolet and visible (UV-Vis) monitoring and high performance liquid chromatography (HPLC). Fructus Arctii should be processed under heating in a pot at 311 °C, medicine at 119 °C for 123s with flipping frequently. The experimental values under the optimized processing technology were consistent with the predicted values. In conclusion, RSM is an effective method to optimize the processing of traditional Chinese medicine (TCM).
Arctium ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Furans ; analysis ; Glucosides ; analysis ; Hot Temperature ; Lignans ; analysis ; Surface Properties ; Technology, Pharmaceutical ; methods

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Arctium ; chemistry ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Furans ; isolation & purification ; pharmacology ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Ionomycin ; pharmacology ; Lectins, C-Type ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; T-Lymphocytes ; cytology ; drug effects ; immunology ; Tetradecanoylphorbol Acetate ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the correlation between ITS sequence of Arctium lappa and Fructus Arctii quality of different origin.

METHODThe samples of Fructu arctii materials were collected from 26 different producing areas. Their ITS sequence were determined after polymerase chain reaction (PCR) and quality were evaluated through the determination of arctiin content by HPLC. Genetic diversity, genotype and correlation were analyzed by ClustalX (1.81), Mage 4.0, SPSS 13.0 statistical software.

RESULTITS sequence of A. was obtained from 26 samples, and was registered in the GenBank. Corresponding arctiin content of Fructus arctii and 1000-grain weight were determined. A. lappa genotype correlated with Fructus arctii quality by statistical analysis.

CONCLUSIONThe research provided a foundation for revealing the molecular mechanism of Fructus arctii geoherbs.

Arctium ; chemistry ; genetics ; Computational Biology ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; standards ; Furans ; analysis ; Genotype ; Glucosides ; analysis ; Polymorphism, Genetic ; genetics ; Quality Control ; Sequence Analysis, DNA ; Software

OBJECTIVETo investigate the reactive oxygen species (ROS) content and nuclear transcription factor-kappa B expression (NF-kappaB) in renal tissue of diabetic rats and the effect of Astragalus and Arctium in combination on them.

METHODSStreptozotocin (STZ)-induced diabetic model rats were established. Astragalus and Arctium in different dosages (low, moderate, high) were combined after orthogonal design and administered respectively to the model rats via gastrogavage for 4 or 8 weeks. Content of ROS in the renal tissue was detected by flow cytometry and expression of functional NF-kappaB p65 was assessed by Western blot at the 4th and 8th weekends (4th week and 8th week).

RESULTSROS content of renal tissue in the model rats was 36.55 +/- 7.46% at the 4th week and 31.91 +/- 5.83% at the 8th week, NF-kappaB p65 expression was 165.00 +/- 3.14 at the 4th week and 214.00 +/- 5.11 at the 8th week, all higher than those in normal rats (6.21 +/- 1.83% and 129.00 +/- 1.58 at the 4th week, 6.95 +/- 1.41% and 148.00 +/- 2.32 at the 8th week) respectively. The combined use of Astragalus and Arctium showed decreasing effects on both indexes significantly, and the decreasing effects of the combination with moderate and high dose Astragalus were better than those with low dose, with the details as follows: those of ROS at the 4th week were 11.43 +/- 2.42%, 18.37 +/- 7.58% and 22.10 +/- 4.71% for high, moderate and low dose Astragalus combination respectively (same hereinafter), at the 8th week 12.55 +/-4.40%, 19.15 +/- 6.64% and 23.48 +/- 3.13%; and for NF-kappaB p65 expression at the 8th week, 185.00 +/- 6.99, 183.00 +/- 3.89 and 194.00 +/- 4.98 respectively.

CONCLUSIONCombined use of Astragalus and Arctium may ameliorate the condition of diabetic nephropathy by inhibiting the activation of the ROS-NF-kappaB signal passage.

Animals ; Arctium ; chemistry ; Astragalus Plant ; chemistry ; Diabetic Nephropathies ; chemically induced ; drug therapy ; metabolism ; Disease Models, Animal ; Drug Therapy, Combination ; Gene Expression ; drug effects ; Humans ; NF-kappa B ; genetics ; metabolism ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Streptozocin ; adverse effects

This study investigated the effect of arctigenin (ARG) on the induction of apoptosis and the putative pathways of its action in HL-60 and K562 cells. MTT assay was used to detect the cytotoxic effect of ARG in HL-60 and K562 cells. The apoptosis was observed by Hoechst 33258 fluorescence staining and DNA agarose gel electrophoresis. Caspase-3 enzyme activity was measured by caspase-3 enzyme activity detection kit. The expression of related protein was analyzed by Western blotting and the vascular endothelial growth factor (VEGF) level was determined by enzyme-linked immunosorbent assay (ELISA). ARG-treated HL-60 cells and K562 cells exhibited growth inhibition and displayed several features of apoptosis, including DNA fragmentation and DNA laddering by agarose gel electrophoresis. It was observed that poly-(ADP-ribose) polymeras (PARP) were cleaved to smaller molecules and ARG induced upregulation of bax and downregulation of bcl-2 protein expression. However, it had no effect on VEGF levels. Taken together, this study demonstrated that ARG is a potent inducer of apoptosis and this was accompanied by caspase-3 activation and upregulation of bax/bcl-2, which offers a potential mechanism for the apoptosis-inducing activity of ARG.
Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Arctium ; chemistry ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; DNA Fragmentation ; drug effects ; Fruit ; chemistry ; Furans ; isolation & purification ; pharmacology ; HL-60 Cells ; Humans ; K562 Cells ; Lignans ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

HPLC and LC-MS/MS were used to establish a comprehensive HPLC analytical method of Yinqiao decoction and identify the chemical constituents of the whole and individual herbs of Yinqiao decoction. YWG-C18 (250 mm x4. 6 mm ID, 10 microm) column was used; the mobile phase was composed of acetonitrile (A) and water ( B, with 3% acetic acid) with gradient elution; the flow rate was 1. 0 mL x min(-1) and the column temperature was set up at 25 degrees C. The detection wavelength was 280 nm. The chromatographic fingerprints of Yinqiao Decoction showed 30 main peaks. Peak 2, 14, 15, 17 were from Lonicera japonica Thunb, peak 3, 12, 13, 24 were from Fosythia suspense (Thunb) Vahl, peak 19, 25, 26, 27 were from Arctium lappa L. , peak 5, 6, 8, 9, 10, 11, 18, 28 were from Glycyrrhiza uralensis Fisch, peak 20, 21 were from Mentha haplocalyx Briq. , peak 22, 23 were from Schizonepeta tenuifolia Briq. , peak 1 presented in the chromatograms of Lonicera japonica Thunb, Fosythia suspense (Thunb) Vahl, Mentha haplocalyx Briq. , Schizonepeta tenuifolia Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 7 presented in the chromatograms of Fosythia suspense (Thunb) Vahl and Glycine max (L. ) Merr. , peak 16 presented in the chromatograms of Mentha haplocalyx Briq. and Schizonepeta tenuifolia Briq. , peak 29 presented in the chromatograms of the herbs except Mentha haplocalyx Briq. and Platycodon grandiflorum (Jacq. ) A. DC. , peak 30 presented in the chromatograms of the herbs except Platycodon grandiflorum (Jacq. ) A. DC. , peak 4 was not identified, maybe it was a new constituent produced during decoction. By comparison of the standards isolated and MS spectra, 14 peaks were identified as 2 ( chlorogenic acid) , 9 ( liquiritin ) , 10 ( 4'-O-[ beta-D-apiofuranosyl (1--> 2 ) -beta-D-glucopyranosyl] liquiritigenin), 12 (forsythiaside), 13 (rutin), 14 (4,5-O-dicaffeoylquiniic acid), 15 (3, 5-O-dicaffeoylquiniic acid ), 16 ( 4-0- [ beta-D-apiofuranosyl ( 1 -->2 ) -beta-D-glucopyranosyl ] isoliquiritigenin) , 17 ( 3, 4-O-dicaffeoylquiniic acid) , 18 (2'-O-[ beta-D-apiofuranosyl (1 -->2 ) -beta-D-glucopyranosyl] isoliquiritigenin) , 19 (arctiin) , 20 (linarin) , 25 (genistein) , 28 ( isoliquiritigenin) . The method could be used to identify the characteristics of Yinqiao decoction, and it could be used to evaluate the quality and quantity of Yinqiao decoction.
Arctium ; chemistry ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavanones ; analysis ; Forsythia ; chemistry ; Furans ; analysis ; Glucosides ; analysis ; Glycosides ; analysis ; Glycyrrhiza uralensis ; chemistry ; Lonicera ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; instrumentation ; methods

OBJECTIVETo study the different manufacturers Vitmin C (Vc) Yinqiao tablets, and the quality of the analysis of the problem, to provide a theoretical basis for the correct evaluation of the quality of medicines and improving the standard drugs.

METHOD11 manufacturers of 18 batches of samples for determination of the weight of the core tablets, powder samples were observed with microscope, determination of Vc, and the establishment of the Vc Yinqiao tablets HPLC method for determination of chlorogenic acid and arctigenin, chlorogenic acid and arctigenin in the samples were measured and compared.

RESULTThere is a big difference of microscope and various measured results in different manufacturers products.

CONCLUSIONBecause different manufacturers to produce the same, there are big differences in the quality of the products.

Arctium ; chemistry ; Ascorbic Acid ; chemistry ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Furans ; analysis ; Lignans ; analysis ; Lonicera ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Tablets