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OBJECTIVE To investigate the effect of arctiin （ARC）relieving lipopolysaccharide （LPS）induced inflammatory injury of human nasopharyngeal epithelial cells NP- 69. METHODS The effects of 24 h treatment of 0.000 1，0.001，0.01，0.1， 1.0，10 μmol/L ARC on the proliferation of NP-69 were determined by MTS method. After 0.01，0.1，and 1.0 μmol/L ARC was applied to NP- 69 for 24 h and NP- 69 was pre-treated with 0.01，0.1 and 1.0 μmol/L ARC for 24 h，and then stimulated with 1.0 μg/mL LPS for 24 h，scratch tests were used to detect cell migration in both experiments. LPS stimulated NP- 69 to establish an inflammation injury model. The levels of nitric oxide （NO），tumor necrosis factor α（TNF-α）interleukin-6（IL-6），and IL- 1β in cell supernatants were detected ，and mRNA and protein expression of zonula oecludens protein 1（ZO-1），β-defensin 3（BD3）， Janus kinase 1 （JAK1），signal transducer and activator of transcription 3 （STAT3） in cell supernatant were also detected. RESULTS Compared with normal group ，0.000 1，0.001，0.01，0.1，1.0，10 μmol/L ARC had no effect on the proliferation of NP-69 after 24 h treatment （P＞0.05）. ARC （0.1，1.0 μmol/L）could significantly promote the rate of cell migration （P＜0.05）. For the inflammatory injure of NP- 69 cells stimulated by LPS ，ARC（1.0 μmol/L）could significantly reduce the release of NO ， TNF-α and IL-6（P＜0.05），significantly increased mRNA and protein expression of ZO- 1 and BD 3 but decreased mRNA and protein expression of STAT 3（P＜0.01 or P＜0.05）. CONCLUSIONS ARC has the effect of reducing the inflammatory injury of NP-69 cells induced by LPS ，promoting the physical and immune defense ability of the nasal mucosa epithelial barrierunder inflammatory environment. The mechanism of action may be related to inhibiting IL- 6/JAK1/STAT3 signaling pathway.
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OBJECTIVE@#To investigate the protective effect of arctiin with anti-inflammatory bioactivity against triptolide-induced nephrotoxicity in rats and explore the underlying mechanism.@*METHODS@#Forty SD rats were divided into 4 groups for gastric lavage of normal saline, arctiin (500 mg/kg), triptolide (500 μg/kg), or both arctiin (500 mg/kg) and triptolide (500 μg/kg). Blood samples were collected for analysis of biochemical renal parameters, and the renal tissues were harvested for determining the kidney index and for pathological evaluation with HE staining. In the @*RESULTS@#In SD rats, arctiin significantly antagonized triptolide-induced elevation of BUN, Scr and kidney index (@*CONCLUSIONS@#Arctiin can protect the kidney from triptolide-induced damages in rats possibly through the anti-inflammatory pathway.
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Animals , Rats , Anti-Inflammatory Agents , Diterpenes/toxicity , Epoxy Compounds/toxicity , Furans , Glucosides , Kidney/drug effects , Phenanthrenes/toxicity , Rats, Sprague-DawleyABSTRACT
In this study, we accurately collected the embryonic parenchyma cells and endocarp stone cells of Arctii Fructus at five different growth stages by laser microdissection. Quantitative analyse of caffeic acid, arctiin and arctigenin in these cells were performed using ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS). The results showed that a large amount of arctiin was produced and accumulated in embryonic parenchyma cells from the late flowering stage to mature stage, while much lower content of arctiin was produced and accumulated in endocarp stone cells at these stages. It suggested that the biosynthetic pathways of arctiin were different in embryonic parenchyma cells from endocarp stone cells of Arctii Fructus. Arctigenin was found to be produced and accumulated in both embryonic parenchyma cells and in endocarp stone cells from the late flowering stage to mature stage, but it reached a peak in endocarp stone cells at late flowering stage, then decreased slowly. The concentration of arctigenin was far less than that of arctiin regardless of embryonic parenchyma cells or endocarp stone cells. These results have validated the new method for analysis of dynamic accumulation of arctiin in Arctii Fructus by UFLC-MS/MS with frozen sections and microdissection.
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OBJECTIVE:To establish a method for the simultaneous determination of rutin ,hesperidin ,arctiin ,liquiritin and glycyrrhizinate in Sangxing oral liquid. METHODS:RP-HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.1% phosphoric acid solution(gradient elution) at a flow rate of 1.0 mL/min,the detection wavelength was 276, 348 nm ,column temperature was 30 ℃,and the injection volume was 5 μL. RESULTS:The linear range was 1.068-10.68 μg/mL (r=0.9993)for rutin,9.320-93.20 μg/mL(r=0.9992)for hesperidin,46.91-469.1 μg/mL(r=0.9994)for arctiin,0.5110-5.110μg/mL(r=0.9992) for liquiritin;RSDs of precision , stability and reproducibility tests were less than 3%;recoveries were 95.10% -101.90%(RSD=2.1%,n=9),96.75%-101.80%(RSD=1.4%,n=9),95.69%-100.00%(RSD=1.5%,n=9), 98.22%-104.60%(RSD=2.1%,n=9),respectively. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the simultaneous determination of rutin,hesperidin,arctiin and liquiritin in Sangxing oral liquid.
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OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.
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Objective: To establish a method for determining the contents of hesperidin and arctiin in the root of Arctium lappa L.from different habitats.Methods: The chromatographic separation was performed on an Acquity UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm).The mobile phase consisted of acetonitrile (A)-0.1% formic acid (B) with gradient elution at the flow rate of 0.3 ml·min-1.The detection wavelength was 280 nm, and the temperature of column was 25℃.Results: The linear range of hesperidin and arctiin was 4.950-29.700 μg·ml-1 and 6.250-37.500 μg·ml-1(r=0.999 9), respectively.The average recovery of hesperidin and arctiin was 99.0% and 98.5%, respectively.Conclusion: The developed method is sensitive, rapid and accurate, which can be used for the quality control of the root,of Arctium lappa L.
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Objective: To establish HPLC fingerprint for Yinqiao Qingre Tablet (YQT), and analyze the chemical composition of YQT by ESI-Q-TOF MS. Methods: The Kromasil C18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of acetonitrile-0.1% phosphoric acid in gradient elution, the flow rate was 1.0 mL/min, the column temperature was 25℃, and the detection wavelength was 230 nm. Ultra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was used for qualitative analysis in the positive and negative ion modes. Results: The HPLC fingerprint for 11 batches of YQT and 28 common peaks were obtained, belonging to six medicinal herbs. Among them, seven common peaks came from Forsythiae Fructus, nine common peaks came from Lonicerae Japonicae Flos, six common peaks came from Puerariae Lobatae Radix, three common peaks came from Arctii Fructus, one common peak (peak 2) came from Forsythiae Fructus and Cimicifuga Rhizoma, another two common peaks came from Anemarrhenae Rhizoma and Cimicifuga Rhizoma, respectively. The similarity of 11 batches of YQT was over 0.95.Totally 42 chemical components were identified by UPLC-Q-TOF/MS, 16 of which were identified by references, such as puerarin, daidzin, daidzein, neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, secoxyloganin, rutin, phillyrin, forsythoside A, arctiin, mangiferin, and timosaponin B II, respectively. Conclusion: This study perfects the system of quality standard and provides the reference for the study of substance basis and quality control of the same type of compound preparations.
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Objective: To study the difficult extraction technology of Arctii Fructus in separation of arctigenin industrialization, which provides a practical preparation technology suitable for arctigenin industrialization in order to promote the industrialization development of arctigenin. Methods: Arctii Fructus was hydrolyzed by acid hydrolysis and alcohol extraction. Then the crude product was separated by ethyl acetate extraction. Finally the pure product of arctigenin was obtained through ethanol crystallization. Results: We obtained the crude product which the purity was more than 75.0% by simple extraction and separation. The finished product was crystallized with anhydrous ethanol until the purity of arctigenin is more than 99.0%. Conclusion: This study obtains a suitable process for arctigenin industrialization preparation.
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This study was aimed to establish a simultaneous determination method of chlorogenic acid,liquiritin,rosmarinic acid,arctiin,glycyrrhizic acid,schisandrin,menthone in Guo-Ming-Xing Bi-Yan(GMXBY) granules by HPLC under multiple UV wavelengths.Waters Symmetry C18 column (4.6 mm× 250 mm,5μm) was used as the chromatographic column.Acetonitrile-0.1% phosphoric acid water solution was used as the mobile phase with gradient elution.The detection wavelength of chlorogenic acid and rosmarinic acid was 327 nm; that of liquiritin and arctiin was 280 nm; that of glycyrrhizic acid,schisandrin and menthone was 250 nm.The column temperature was 25℃.The injection volume was 10μL.Chlorogenic acid showed a good linear relationship in the range of 1.19-59.50μg?mL-1 (r = 0.999 8).The average recovery rate was 100.95%.Liquiritin showed a good linear relationship in the range of 1.51-150.70μg?mL-1 (r = 0.999 1).The average recovery rate was 100.38%.Rosmarinic acid showed a good linear relationship in the range of 3.40-68.08 μg?mL-1 (r = 0.999 9).The average recovery rate was 101.02%.Arctiin showed a good linear relationship in the range of 56.15-1 123.00μg?mL-1 (r =0.999 9).The average recovery rate was 100.39%.Glycyrrhizic acid showed a good linear relationship in the range of 21.54-430.80μg?mL-1 (r = 0.999 8).The average recovery rate was 97.09%.Schisandrin showed a good linear relationship in the range of 2.57-51.34μg?mL-1 (r = 0.999 9).The average recovery rate was 99.19%.Menthone showed a good linear relationship in the range of 0.50-10.00μg?mL-1 (r = 0.999 9).The average recovery rate was 100.35%.This established method was simple and reliable with good reproducibility,which can be used as the determination method of active components in GMXBY granules.
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This study was aimed to optimize the extraction process of double-marker components for Arctium lappa L. The central composite design and response surface methodology was used. According to 3 main factors, the extraction rates of arctiin and arctigenin was used as evaluation indexes. Multiple linear regression and two-order polynomial equation were used. The binomial fitting model was performed in the optimization of arctiin and arctigenin extraction technology. The results showed that the indentified optimized extraction technology of arctiin and arctigenin was 70% ethanol, 24-fold, ultrasonic solvent extraction for 15 minutes. It was concluded that this technology was able to extract large amount of arctiin and arctigenin, which provided experiment evidences for arctiin and arctigenin preparation. It also provided references for the development and utilization of arctiin and arctigenin.
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Objective:To investigate the hpyerglycemic action of arctiin in db/db mice with spontaneous diabetes and the underly-ing mechanism. Methods:Totally 40 db/db mice were randomly divided into five groups: the model control group, arctiin group re-spectively with the dose of 75, 150, 300 mg·kg-1 , 300 mg·kg-1 metformin group. The age-matched db/m mice were selected as the normal control group. The mice were administered with corresponding drugs or solvent by gavage for 4 weeks. The oral glucose tol-erance test was carried out at the end of the 3rd week. After the 4-week treatment, all the mice were fasted overnight (12h), and then the body weight and fasting blood glucose ( FBG ) were determined. The concentration of insulin ( INS ) , glycated serum protein ( GSP) , triglyceride ( TG) , total cholesterol ( TC) and adiponection ( APN) were detected. Results:Arctiin could significantly lower the body weight and FBG, improve the glucose tolerance, decrease the serum concentration of INS, GSP, TG, TC and APN(P<0. 05 or 0. 01). Conclusion:Arctiin has benefit effects against glucose/lipid metabolism disorder and insulin resistance in db/db diabetic mice. The mechanism may be related to up-regulating the expression of adiponection.
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Objective: To establish a high performance liquid chromatographic method for simultaneous determination of rutin, hesperidin, and arctiin in Fupo Ganmao Granules. Methods: HPLC was applied with the chromatographic condition as follows: The chromatographic column was Agilent Zorbax SB-C18 (250 mm × 4.6 mm, 5 μm) at 30°C; methanol-water as the mobile phase, gradient elution; flow rate was 0.8 mL/min and the detection wavelength was 280 nm. Results: Linear relationships were 5.204-130.1 μg/mL (r = 0.9990), 1.819-145.5 μg/mL (r = 1.0000), and 43.22-778.0 μg/mL (r = 0.9999), respectively. The average recoveries were 98.77% (RSD = 1.69%), 99.52% (RSD = 1.01%), and 100.84% (RSD = 1.59%). Conclusion: The results show that the method is simple, accurate, and repeatable, and it is suitable for the simultaneous determination of rutin, hesperidin, and arctiin in Fupo Ganmao Granules.
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BACKGROUND: Accumulating evidence indicates that reactive oxygen species (ROS) are an important etiological factor for the induction of dermal papilla cell senescence and hair loss, which is also known alopecia. Arctiin is an active lignin isolated from Arctium lappa and has anti-inflammation, anti-microbial, and anti-carcinogenic effects. In the present study, we found that arctiin exerts anti-oxidative effects on human hair dermal papilla cells (HHDPCs). RESULTS: To better understand the mechanism, we analyzed the level of hydrogen peroxide (H2O2)-induced cytotoxicity, cell death, ROS production and senescence after arctiin pretreatment of HHDPCs. The results showed that arctiin pretreatment significantly inhibited the H2O2-induced reduction in cell viability. Moreover, H2O2-induced sub-G1 phase accumulation and G2 cell cycle arrest were also downregulated by arctiin pretreatment. Interestingly, the increase in intracellular ROS mediated by H2O2 was drastically decreased in HHDPCs cultured in the presence of arctiin. This effect was confirmed by senescence associated-beta galactosidase (SA-ß-gal) assay results; we found that arctiin pretreatment impaired H2O2-induced senescence in HHDPCs. Using microRNA (miRNA) microarray and bioinformatic analysis, we showed that this anti-oxidative effect of arctiin in HHDPCs was related with mitogen-activated protein kinase (MAPK) and Wnt signaling pathways. CONCLUSIONS: Taken together, our data suggest that arctiin has a protective effect on ROS-induced cell dysfunction in HHDPCs and may therefore be useful for alopecia prevention and treatment strategies.
Subject(s)
Humans , Aging/metabolism , Reactive Oxygen Species/antagonists & inhibitors , Hair Follicle/drug effects , MicroRNAs/metabolism , Furans/pharmacology , Glucosides/pharmacology , Aging/drug effects , Down-Regulation/drug effects , Up-Regulation/drug effects , Cell Line , Cell Survival/drug effects , Cell Death/drug effects , beta-Galactosidase/analysis , Hair Follicle/cytology , Hair Follicle/metabolism , Dermis/cytology , Dermis/drug effects , Dermis/metabolism , Oligonucleotide Array Sequence Analysis , MicroRNAs/drug effects , Cell Cycle Checkpoints/drug effects , Hydrogen Peroxide/pharmacologyABSTRACT
BACKGROUND/OBJECTIVES: The purpose of this study was to examine the effects and associated mechanisms of arctiin, a lignan compound found in burdock, on adipogenesis in 3T3-L1 cells. Also, the effects of arctiin supplementation in obese mice fed a high-fat diet on adiposity were examined. MATERIALS/METHODS: 3T3-L1 cells were treated with arctiin (12.5 to 100 microM) during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining and intracellular triglyceride contents. The expressions of genes related to adipogenesis were measured by real-time RT-PCR and Western blot analyses. For in vivo study, C57BL/6J mice were first fed either a control diet (CON) or high-fat diet (HF) to induce obesity, and then fed CON, HF, or HF with 500 mg/kg BW arctiin (HF + AC) for four weeks. RESULTS: Arctiin treatment to 3T3-L1 pre-adipocytes markedly decreased adipogenesis in a dose-dependent manner. The arctiin treatment significantly decreased the protein levels of the key adipogenic regulators PPARgamma and C/EBPalpha, and also significantly inhibited the expression of SREBP-1c, fatty acid synthase, fatty acid-binding protein and lipoprotein lipase. Also, arctiin greatly increased the phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target phosphorylated-acetyl CoA carboxylase. Furthermore, administration of arctiin significantly decreased the body weight in obese mice fed with the high-fat diet. The epididymal, perirenal or total visceral adipose tissue weights of mice were all significantly lower in the HF + AC than in the HF. Arctiin administration also decreased the sizes of lipid droplets in the epididymal adipose tissue. CONCLUSIONS: Arctiin inhibited adipogenesis in 3T3-L1 adipocytes through the inhibition of PPARgamma and C/EBPalpha and the activation of AMPK signaling pathways. These findings suggest that arctiin has a potential benefit in preventing obesity.
Subject(s)
Animals , Mice , 3T3-L1 Cells , Adenylate Kinase , Adipocytes , Adipogenesis , Adipose Tissue , Adiposity , AMP-Activated Protein Kinases , Blotting, Western , Body Weight , Diet , Diet, High-Fat , Intra-Abdominal Fat , Lipoprotein Lipase , Mice, Obese , Obesity , Phosphorylation , PPAR gamma , Sterol Regulatory Element Binding Protein 1 , Triglycerides , Weights and MeasuresABSTRACT
OBJECTIVE: To establish a method for the determination of chlorogenic acid, phillyrin, arctiin, liquiritin and glycyrrhizin in Yinqiao Powder. METHODS: High performance liquid chromatography was performed on a Kromasil C18 column (4.6 mm × 250 mm, 5 μm) at 30°C. The mobile phase was acetonitrile -0.4% phosphoric acid solution with gradient elution. The flow rate was 1.0 mL · min-1 and the detection wavelength was set at 230 nm. RESULTS: Chlorogenic acid, phillyrin, arctiin, liquiritin and glycyrrhizin had good liner relationship in the ranges of 0.0394-0.788, 0.0205-0.410, 0.068-1.36, 0.0348-0.697 and 0.0376-0.752 μg, respectively. The correlation coefficients were all more than 0.9996. The average recoveries were 101.3% (RSD=1.5%), 99.5% (RSD=1.2%), 98.7% (RSD=1.9%), 96.7% (RSD=2.3%) and 98.2% (RSD=2.0%), respectively. CONCLUSION: This method is convenient, rapid, accurate, reliable and has good reproducibility, which can be used for the determination of chlorogenic acid, phillyrin, arctiin, liquiritin and glycyrrhizin in Yinqiao Powder. This study provides scientific basis for the quality evaluation of Yinqiao Powder and preparations of the same kind.
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Objective: To investigate the in vivo absorptive characteristic of arctiin in small intestine of rats. Methods: The intestine absorption model of arctiin was established by in vivo intestinal perfusion method. The absorption rate constants (Ka), absorption half time (t1/2), absorption percent of per unit of time (P), and apparent permeability coefficient (Papp) were calculated and compared to investigate the absorptive dynamics of arctiin in intestine of rats. Results: At the concentration range of 10-50 μg/mL, Ka, t1/2, P, and Papp showed no significant difference, and had no connection with concentration of arctiin. Conclusion: Arctiin complies with the first-order absorptive kinetics and the absorptive mechanism in intestine may be passive transport.
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Ultrasound-assisted extraction is evaluated as a simpler and more effective alternative to conventional extraction methods for the extraction of bioactive compounds from natural product. This study investigated the use of ultrasound-assisted extraction to extract three dibenzylbutyrolactone lignans, including tracheloside, hemislienoside, and arctiin from Hemistepta lyrata. Factors such as extraction solvent, solvent concentration, solvent to material ratio, and extraction time were examined. High-performance liquid chromatography with photodiode array detection was used for simultaneous determination of the target compounds in the corresponding extracts. Results showed that the optimal parameters to extract the target compounds from H. lyrata were as follows: extraction solvent: 70% aqueous ethanol; solvent to material ratio: 20:1 (v/w, ml/g); extraction time: 20 min under the conditions: ultrasonic frequency: 40 Hz; extraction temperature: 30 ˚C. With all these merits, ultrasound-assisted extraction should be considered for wider application in the extraction of tracheloside, hemislienoside, and arctiin from other medicinal plants.
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Objective To establish a method for the determination of arctiin in Pifu Zhiyang Tablets by SPE-HPLC . Methods SPE-HPLC method was performed on a Kromasil C18 column(150 mm?4.6 mm ,5 ?m),the mobile phase consisted of methanol -1 %acetic acid (35 ∶65) ,the wavelength was 283nm . Results The linear range of arctiin was 1.5～4.5 ?g(r= 0. 999 8) ,the average recovery was 98.18 %with RSD of 2.08 %. Conclusion This method is accurate ,reliable and reproducible. It can be used for the determination of arctiin in Pifu Zhiyang Tablets .
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AIM: To investigate and optimize the technology of ultrasonic-assisted extraction of arctiin and arctigenin from Fructus arctii.METHODS: The effects of ultrasonic power,particle size,ratio of liquid to solid,extraction time,extraction temperature and duty factor on the extraction rates of arctiin and arctigenin were investigated,and optimized with orthogonal experiments.RESULTS: The results showed that the optimal extraction conditions were as follows: particle 80 ～ 100 mesh 14 mL/g,ratio of liquid to solid 400 W ultrasonic power,50 ℃ of extraction temperature,and 10 min of extraction time.CONCLUSION: The optimized conditions are reasonable and reliable.
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AIM: To optimize the parameters of processing for Arctium Lappa.METHODS: The contents of arctiin and arctigenin in Arctium Lappa were determined with HPLC,and the total yields of them were chosen as in-dex.The orthogonal test was adopted to acquire the optimum technology for processing Arctium Lappa.RESULTS: The study showed that the best processing parameters for Arctium Lappa consisted of 300 ℃ of the drying tempera-ture and 4-5 minutes of processing time.CONCLUSION: The processing method is reasonable,stable and reliable,and can be used in commercial production.