Arctigenin mitigates vascular endothelial injury in rats with pregnancy-induced hypertension via autophagy-NLRP3 inflammasome pathway / 中国中药杂志

This study aims to investigate the effect and mechanism of arctigenin(ARC) in the treatment of vascular endothelial injury in rats with pregnancy-induced hypertension(PIH). Fifty SD rats pregnant for 12 days were randomly assigned into a control group, a model group, an ARC group, a rapamycin(RAP, autophagy inducer) group, and an ARC+3-methyladenine(3-MA, autophagy inhibitor) group, with 10 rats in each group. The rats in the other groups except the control group were intraperitoneally injected with nitrosyl-L-arginine methyl ester(50 mg·kg~(-1)·d~(-1)) to establish the PIH model on the 13th day of pregnancy. On the 15th day of pregnancy, the rats in ARC, RAP, and ARC+3-MA groups were intraperitoneally injected with ARC(50 mg·kg~(-1)·d~(-1)), RAP(1 mg·kg~(-1)·d~(-1)), and 3-MA(15 mg·kg~(-1)·d~(-1))+ARC(50 mg·kg~(-1)·d~(-1)), respectively. The pregnant rats in the control group and the model group were intraperitoneally injected with the same amount of normal saline. The blood pressure and 24 h urine protein(24 h-UP) of pregnant rats in each group were measured before and after intervention. Cesarean section was performed to terminate pregnancy on day 21, and the body weight and body length of fetal rats were compared among groups. Hematoxylin-eosin(HE) staining was employed to observe the pathological changes of placenta. The expression of endothelin-1(ET-1) and endothelial nitric oxide synthase(eNOS) in placenta was detected by immunohistochemistry. The serum levels of ET-1 and nitric oxide(NO) were determined with corresponding kits. The expression of microtubule-associated protein 1 light chain 3(LC3), Beclin-1, NOD-like receptor protein 3(NLRP3), apoptosis-associated speck-like protein with CARD domain(ASC), caspase-1, interleukin(IL)-1β, and IL-18 was determined by immunofluorescence and Western blot. The level of reactive oxygen species(ROS) in placenta was measured by fluorescence staining. The results showed that on day 12 of pregnancy, the blood pressure and 24 h-UP had no significant differences among groups. On days 15, 19, and 21, the blood pressure and 24 h-UP in the model group were higher than those in the control group(P<0.05). On days 19 and 21, the blood pressure and 24 h-UP in ARC group and RAP group were lower than those in the model group(P<0.05), and they were higher in the ARC+3-MA group than in the ARC group(P<0.05). On day 21, the model group had lower body weight and body length of fetal rats(P<0.05), higher serum level of ET-1, and lower serum level of NO(P<0.05) than the control group. Moreover, the placental tissue showed typical pathological damage, down-regulated expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and eNOS(P<0.05), up-regulated expression of ET-1, NLRP3, ASC, caspase-1, IL-1β, and IL-18(P<0.05), and elevated ROS level. Compared with the model group, ARC and RAP groups showed increased body weight and body length of fetal rats(P<0.05), lowered serum level of ET-1, elevated serum level of NO(P<0.05), reduced pathological damage of placental tissue, up-regulated expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, and eNOS(P<0.05), down-regulated expression of ET-1, NLRP3, ASC, caspase-1, IL-1β, and IL-18(P<0.05), and lowered ROS level. Compared with ARC group, 3-MA reversed the effects of ARC on the above indicators. In conclusion, ARC can inhibit the activation of NLRP3 inflammasome and mitigate vascular endothelial damage in PIH rats by inducing autophagy of vascular endothelial cells.

Arctigenin regulates the effect of miR-21 on hypoxia-reoxygenation-induced pyroptosis of cardiomyocytes / 西安交通大学学报(医学版)

【Objective】 To investigate the effect and mechanism of arctigenin (ARG) on hypoxia-reoxygenation (H/R) induced pyroptosis of cardiomyocytes. 【Methods】 H9C2 cells were cultured in vitro, and underwent hypoxia for 2 hours and reoxygenation for 4 hours to establish H/R cell injury model. The cells were divided into control group (Control), model group (H/R), ARG group, miR-21 simulation group (miR-21 mimic), and ARG+miR-21 inhibitor group (ARG+miR-21 inhibitor). TUNEL staining was used to detect the pyroptosis index of H9C2 cells; the lactate dehydrogenase (LDH) kit was used to detect the release of LDH in each group of cells; the enzyme-linked immunosorbent assay (ELISA) was used to detect the content of interleukin-1β (IL-1β) and interleukin-18 (IL-18). Western blotting was used to detect the expressions of pyroptosis-related proteins (Caspase-1, GSDMD, IL-1β and IL-18) in each group. 【Results】 Compared with those in the control group, the pyroptosis index, the release of LDH, IL-1β and IL-18, and the protein expressions of Caspase-1p20, GSDMD-N, IL-1β and IL-18 in the H/R group were significantly increased (P<0.01). Compared with H/R group, ARG group and miR-21 mimic group had significantly reduced pyroptosis index, LDH, IL-1β and IL-18 release, and protein expressions of Caspase-1 p20, GSDMD-N, IL-1β and IL-18 (P<0.01), and the above-mentioned index changes could be reversed after treatment with +miR-21 inhibitor. 【Conclusion】 ARG can inhibit H/R-induced pyroptosis of cardiomyocytes, and its mechanism is related to the promotion of miR-21 expression.

Arctigenin attenuates paraquat-induced human lung epithelial A549 cell injury by suppressing ROS/p38 mitogen-activated protein kinases-mediated apoptosis / 世界急诊医学杂志（英文）

@#BACKGROUND: Paraquat (PQ)-induced acute lung injury (ALI) and pulmonary fibrosis are common diseases with high mortality but without effective antidotes in emergency medicine. Our previous study has proved that arctigenin suppressed pulmonary fibrosis induced by PQ. We wondered whether arctigenin could also have a protective effect on PQ-induced ALI. METHODS: A PQ-induced A549 cell injury model was used, and the effect of arctigenin was determined by a cell counting kit-8 (CCK-8) cell viability assay. In addition, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) staining assays and mitochondrial membrane potential assays were performed to evaluate the level of cell apoptosis. The generation of reactive oxygen species (ROS) was reflected by dihydroethidium (DHE) staining and a 2’,7’-dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Moreover, immunoblotting studies were used to assess the expression of mitogen-activated protein kinases (MAPKs) and p38 MAPK. RESULTS: Arctigenin attenuated PQ-induced inhibition of A549 cell viability in a dose-dependent manner. Arctigenin also significantly reduced PQ-induced A549 cell apoptosis, as reflected by the TUNEL assay and mitochondrial membrane potential assay, which may result from suppressed ROS/p38 MAPK signaling because we found that arctigenin dramatically suppressed ROS generation and p38 MAPK phosphorylation. CONCLUSION: Arctigenin could attenuate PQ-induced lung epithelial A549 cell injury in vitro by suppressing ROS/p38 MAPK-mediated cell apoptosis, and arctigenin might be considered a potential candidate drug for PQ-induced ALI.

Arctigenin attenuates airway inflammation in asthmatic mice via SIRT1/NLRP3 pathway / 中国药理学通报

Aim To investigate the efficacy of arctigenin on airway inflammation in a mouse model of asthma and the mechanism related to the SIRT1/NLRP3 signaling pathway. Methods Forty female BALB/c mice of clean grade were selected and divided into control group, OVA model group and ATG group (5, 10 and 20 mg · kg

Progress in structural modification of representative lignan compounds / 中草药

Lignans which are polymerized by two or more phenylpropanoids derivatives were mostly found in the xylem and resin of plants. Modern pharmacological studies have shown that lignans have extensive biological activities, such as anti-tumor, anti-HIV, antidiabetics, anti-oxidant, cardiovascular and liver protections and so on. However, owing to the poor solubility of structures, its clinical application is still limited. Therefore, chemical modification methods are usually used by domestic and overseas researchers in order to improve their solubility. In this paper, a series of typical lignans such as arctigenin, podophyllotoxin, schisandrin, magnolol, and honokiol were taken as examples to summarize their structural modification methods and different biological activities, aiming to provide scientific basis for further exploitation and studies of lignans.

Chemical Constituents of Ethyl Acetate Fraction from Cynodon dactylon / 中国药学杂志

OBJECTIVE: To isolate and identify the chemical constituents from Cynodon dactylon. METHODS: The chemical constituents of the alcohol extract of C. dactylon. were isolated and purified by various chromatography methods, and their structures were identified by physical and chemical properties and spectral data. RESULTS: Fourteen compounds were isolated from C. dactylon and their structures were examined by physicochemical characteristics and spectral data and identified as vanillin (1), arctigenin (2), maltol (3), (+)-dehydrovomifoliol (4), (3R, 6R, 7E)-3-hydroxy-4, 7-megastigmadien-9-one (5), (-)-loliolide (6), zhebeiresinol (7), 1-indole-3-carboxaldehyde (8), ferulic acid (9), matairesinol (10), pinoresinol (11), ethyl caffeate (12), traxillagenin (13), and impecylenolide (14). CONCLUSION: Compounds 1-14 are isolated from C. dactylon for the first time.

Chemical constituents from plant of Stelleropsis tianschanica / 中草药

Objective: To study the chemical constituents from the arerial parts of Stelleropsis tianschanica. Methods: The constituents were isolated and purified by silica gel chromatography repeatedly, and the structures were identified by spectra analysis and chemical methods. Results: Thirteen compounds were isolated from S. tianschanica and the structures were identified as (+)-pinoresinol (1), (-)-pinoresinol (2), 3'-desmethylarctigenin (3), arctigenin (4), pluviatolide (5), umbelliferone (6), 4-(3,4- dimethoxybenzyl)-3-(4-hydroxy-3-methoxybenzyl)-tetrahydrofuran-2-ol (7), daphnogitin (8), daphnetone (9), blumenol B (10), loliolide (11), 4'-hydroxyacetophenone (12), and 4'-hydroxybenzoic acid (13). Conclusion: Compounds 1-13 were all obtained for the first time from the plant of S. tianschanica and the genus Stelleropsis Pobed.

A novel arctigenin-containing latex glove prevents latex allergy by inhibiting type I/IV allergic reactions / 中国天然药物

The present study aimed at developing a natural compound with anti-allergic effect and stability under latex glove manufacturing conditions and investigating whether its anti-allergic effect is maintained after its addition into the latex. The effects of nine natural compounds on growth of the RBL-2H3 cells and mouse primary spleen lymphocytes were determined using MTT assay. The compounds included glycyrrhizin, osthole, tetrandrine, tea polyphenol, catechin, arctigenin, oleanolic acid, baicalin and oxymatrine. An ELISA assay was used for the in vitro anti-type I/IV allergy screening; in this process β-hexosaminidase, histamine, and IL-4 released from RBL-2H3 cell lines and IFN-γ and IL-2 released from mouse primary spleen lymphocytes were taken as screening indices. The physical stability of eight natural compounds and the dissolubility of arctigenin, selected based on the in vitro pharnacodynamaic screening and the stability evaluation, were detected by HPLC. The in vivo pharmacodynamic confirmation of arctigenin and final latex product was evaluated with a passive cutaneous anaphylaxis (PCA) model and an allergen-specific skin response model. Nine natural compounds showed minor growth inhibition on RBL-2H3 cells and mouse primary spleen lymphocytes. Baicalin and arctigenin had the best anti-type I and IV allergic effects among the natural compounds based on the in vitro pharmacodynamic screening. Arctigenin and catechin had the best physical stability under different manufacturing conditions. Arctigenin was the selected for further evaluation and proven to have anti-type I and IV allergic effects in vivo in a dose-dependent manner. The final product of the arctigenin-containing latex glove had anti-type I and IV allergic effects in vivo which were mainly attributed to arctigenin as proved from the dissolubility results. Arctigenin showed anti-type I and IV allergic effects in vitro and in vivo, with a good stability under latex glove manufacturing conditions, and a persistent anti-allergic effect after being added into the latex to prevent latex allergy.

Qingfei Xiaoyan Wan, a traditional Chinese medicine formula, ameliorates Pseudomonas aeruginosa-induced acute lung inflammation by regulation of PI3K/AKT and Ras/MAPK pathways

Gram-negative pathogen-induced nosocomial infections and resistance are a most serious menace to global public health. Qingfei Xiaoyan Wan (QF), a traditional Chinese medicine (TCM) formula, has been used clinically in China for the treatment of upper respiratory tract infections, acute or chronic bronchitis and pulmonary infection. In this study, the effects of QF on Pseudomonas aeruginosa-induced acute pneumonia in mice were evaluated. The mechanisms by which four typical anti-inflammatory ingredients from QF, arctigenin (ATG), cholic acid (CLA), chlorogenic acid (CGA) and sinapic acid (SPA), regulate anti-inflammatory signaling pathways and related targets were investigated using molecular biology and molecular docking techniques. The results showed that pretreatment with QF significantly inhibits the release of cytokines (TNF-α and IL-6) and chemokines (IL-8 and RANTES), reduces leukocytes recruitment into inflamed tissues and ameliorates pulmonary edema and necrosis. In addition, ATG was identified as the primary anti-inflammatory agent with action on the PI3K/AKT and Ras/MAPK pathways. CLA and CGA enhanced the actions of ATG and exhibited synergistic NF-κB inactivation effects possibly via the Ras/MAPK signaling pathway. Moreover, CLA is speculated to target FGFR and MEK firstly. Overall, QF regulated the PI3K/AKT and Ras/MAPK pathways to inhibit pathogenic bacterial infections effectively.

Therapeutic effect of arctigenin on carbon tetrachloride-induced liver fibrosis in rats / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of arctigenin(ATG) on liver fibrosis in rats induced by carbon tetrachloride(CCl4)and to explore its underlying mechanism. METHODS Sprague-Dawley rats were randomly divided into six groups:vehicle,ATG 3.0 mg · kg-1 group,CCl4 model group,CCl4+ATG 1.0 and 3.0 mg·kg-1 groups,and CCl4+colchicine(COL)0.1 mg·kg-1(toxicity)group. Liver fibrosis was induced by subcutaneous injection of CCl4 in rats for 8 weeks. ATG and colchicine were administrated ig once a day starting from the fifth week after the CCl4 treatment for 4 weeks subsequent. At the end of the study,glutamic pyruvic transaminase (GPT),glutamic oxaloacetic transaminase(GOT),albumin(ALB),and total bilirubin (TBIL) as well as the contents of hydroxyproline (HYP) in liver tissues were measured. Histopathological changes were observed in the liver tissues using hematoxyline-eosin(HE)and Masson’s trichrome staining. The proliferation of hepatic stellate cells (HSC) and expression of cell cycle-related proteins were assayed by indirect immunofluores?cence staining and Western blotting,respectively. RESULTS Compared with CCl4 model group,ATG 1.0 and 3.0 mg · kg-1 improved the liver function by decreasing serum contents of GPT,GOT and TBIL (P<0.05),and increasing serum content of albumin(P<0.05). Histological results indicated that ATG 1.0 and 3.0 mg · kg-1 alleviated liver damage and reduced the formation of fibrous septa. Moreover, ATG 1.0 and 3.0 mg · kg-1 significantly decreased liver HYP when compared with CCl4 model group(P<0.05). In addition,CCl4-induced proliferation of activated HSC was inhibited by ATG 1.0 and 3.0 mg·kg-1, and this was accompanied by down-regulation of cyclin D1,cyclin-dependent kinase(CDK)2,CDK4, and proliferating cell nuclear antigen (PCNA)(P<0.05),and up-regulation of p27kip1 in activated HSC (P<0.05). CONCLUSION ATG can alleviate hepatic injury and fibrosis induced by CCl4,which is probably associated with suppression of the proliferation of activated HSC.

Research on preparation of arctigenin from Arctii Fructus / 中草药

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.

Chemical constituents from inflorescence bracts of Arctii Fructus / 中草药

Objective: To study the chemical constituents from the inflorescence bracts of Arctii Fructus. Methods: The compounds were isolated and purified by the methods of silica gel column chromatography, HPLC, and recrystallization, and the structures were elucidated by the means of spectral analysis. Results: Twelve compounds were isolated and identified as daucosterol (1), isofouquierol (2), (22E)-5α, 8-epidioxyergosta-6, 22-dien-3β-ol (3), 3β-hydroxy-21, 22-epoxyursa-20(30)-en (4), 3β, 21β-dihydroxy-20(30)-en-taraxastane (5), oleanolic acid (6), arctigenin (7), carthamogenin (8), caffeic acid (9), 4(14)-eudesmene-8α, 11-diol (10), monogynol A (11), and lupeol (12). Conclusion; Compounds 2-3, 5, 6, 10-11 are obtained from the plants of Arctium L. for the first time, and compound 12 is isolated from the inflorescence bracts of Arctii Fructus for the first time.

Optimization of Extraction Process for Arctiin and Arctigenin inArctium lappaL. Based on Central Composite Design and Response Surface Methodology / 世界科学技术-中医药现代化

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.

Effects and primary mechanism of arctigenin in C6 rat glioma / 中国药理学通报

Aim To observe the effect and primary mechanism of arctigenin ( ARG) in C6 rat glioma. At the same time, to investigate the effect of ARG com-bined with temozolomide. Methods C6 glioma rat model was established, and 90 rats were divided into six groups, which were subcutaneously administered with model, low and high ARG (0. 05 and 0. 1 mg· kg-1 , sc) , temozolomide (20 mg·kg-1 , p. o. ) , low ARG combined with temozolomide(TMZ / ARG 0. 05) and high ARG combined with temozolomide ( TMZ /ARG 0. 1 ) . The tumor specimens of brain were col-lected after tumor graft. Proliferating cell nuclear anti-gen ( PCNA ) , glial fibrillary acidic protein ( GFAP ) and CD40 in tumor specimens were determined by im-munohistochemistry. Results ① Compared with the model group, the tumor sizes of rats in the arctigenin treatment groups were decreased ( P significantly decreased PCNA and CD40 expression ( P<0. 05 ) and increased GFAP expression ( P<0. 05 ) .③ Compared with model group, arctigenin combined with temozolomide decreased the tumor sizes ( P <0. 01 ) , and the tumor inhibition rate was higher than that of the arctigenin and temozolomide. At the same time, arctigenin combined with temozolomide de-creased PCNA and CD40 expression ( P <0. 01 ) and increased GFAP expression ( P <0. 05 ) , which was better than arctigenin and temozolomide. Conclusion Arctigenin inhibits rat glioma growth, and synergizes with temozolomide, which may be associated with in-hibiting PCNA and CD40 expression and strengthening GFAP expression.

Pharmacokinetics of arctigenin nanoemulsion in rats / 中国药学杂志

OBJECTIVE: To develop an HPLC method for the determination of arctigenin in rat plasma and study the pharmacokinetics of arctigenin nanoemulsion. METHODS: The plasma samples were extracted by ethyl acetate. The determination was carried out on an Agilent C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase consisting of methanol-0.2% phosphoric acid (52:48, V/V) and the UV detection wavelength was set at 280 nm. Quercetin was selected as internal standard. Arctigenin nanoemulsion was administered to rats by intravenous injection at a dose of 4 mg · kg-1. The plasma concentration of arctigenin at different time points was determined by the established HPLC method. The pharmacokinetic parameters were processed by DAS2.1 software. RESULTS: The calibration curve of arctigenin had acceptable linearity in the range of 0.1-10 mg · L-1 in rat plasma(r=0.9992). The lower limit of quantitation (LLOQ) was estimated to be 0.1 mg · L-1 and the lower limit of detection (LLOD) was estimated to be 0.03 mg · L-1. The mean extraction recovery rates of arctigenin QC samples at low, medium and high concentration levels were all a-bove 85%. The intra-day and inter-day precisions were less than 6%. The pharmacokinetics of arctigenin in rats after intravenous injection were fitted to a two-compartment model and the half-lives of α phase and β phase were (0.134 ± 0.085) and (1.471 ± 0.164) h, respectively. CONCLUSION: The HPLC method is simple, rapid and accurate. It can be used for monitoring the plasma concentration of arctigenin and its pharmacokinetic study. After intravenous administration of arctigenin nanoemulsion, arctigenin is rapidly distributed into tissues, but the elimination is slow.

Effects and its mechanism of Arctigenin on mouse spleen cells / 安徽医科大学学报

Objective To investigate the effects of Arctigenin ( ATG ) on concanavalin ( ConA )-stimulated cell proliferation and cytokine secretion in mouse spleen cells, and its possible mechanism. Methods The toxicity of ATG on mouse spleen cells was determined by MTT assay. The inhibition of proliferation was investigated by tritiat-ed thymidine incorporation method. Secreted cytokines (IFN-γand IL-2) were analyzed by ELISA. The associated proteins and phosphorylation levels of mTOR pathway ( mTOR/P70 S6 K/Akt/AMPK/Raptor ) were detected by Western blot. Results ATG had no significant toxicity to mouse spleen cells. ATG significantly inhibited mouse primary spleen cells proliferation induced by ConA. ATG suppressed IL-2 and IFN-γ production of mouse spleen cells in a concentration-dependent manner. ATG remarkably suppressed the phosphorylation of mTOR and P70S6K, and enhanced the phosphorylation of upstream AMPK and Raptor, while the phosphorylation of Akt did not change significantly. Conclusion ATG markedly suppresses the proliferation of mouse spleen stimulated by ConA cells and secretion of IFN-γand IL-2 , which may be correlated to the abilities of enhancing the phosphoryla-tion of AMPK and Raptor, inhibiting the phosphorylation of mTOR and P70S6K.

Arctigenin Increases Hemeoxygenase-1 Gene Expression by Modulating PI3K/AKT Signaling Pathway in Rat Primary Astrocytes

In the present study, we found that the natural compound arctigenin inhibited hydrogen peroxide-induced reactive oxygen species (ROS) production in rat primary astrocytes. Since hemeoxygenase-1 (HO-1) plays a critical role as an antioxidant defense factor in the brain, we examined the effect of arctigenin on HO-1 expression in rat primary astrocytes. We found that arctigenin increased HO-1 mRNA and protein levels. Arctigenin also increases the nuclear translocation and DNA binding of Nrf2/c-Jun to the antioxidant response element (ARE) on HO-1 promoter. In addition, arctigenin increased ARE-mediated transcriptional activities in rat primary astrocytes. Further mechanistic studies revealed that arctigenin increased the phosphorylation of AKT, a downstream substrate of phosphatidylinositol 3-kinase (PI3K). Treatment of cells with a PI3K-specific inhibitor, LY294002, suppressed the HO-1 expression, Nrf2 DNA binding and ARE-mediated transcriptional activities in arctigenin-treated astrocyte cells. The results collectively suggest that PI3K/AKT signaling pathway is at least partly involved in HO-1 expression by arctigenin via modulation of Nrf2/ARE axis in rat primary astrocytes.

Determination of seven components in Forsythia suspensa by RP-HPLC / 中草药

Objective: To establish an HPLC method for the simultaneous determination of cafferic acid, forsythoside A, forsythoside B, rutin, hyperoside, forsythin, and arctigenin in Forsythia suspensa. Methods: The analysis was carried out on an Inertsil ODS-3 C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase was composed of acetonitrile and 0.2% phosphorie acid aqueous with gradient elution. The detection wavelength was set at 275 nm. The flow rate was 1.0 mL/min at column temperature of 30 °C. Results: Cafferic acid, forsythoside A, forsythoside B, rutin, hyperoside, forsythin, and arctigenin were well separated by this method, and showed a good linearity in the ranges of 18.24-91.20, 5.88-29.40, 132.60-663.00, 8.34-41.70, 1.96-9.80, 7.60-38.00, and 11.34-56.70 μg/mL, respectively. The average recoveries of the seven components were 97.7%, 96.7%, 102.6%, 101.3%, 93.2%, 91.8%, and 96.7% and the RSD values were 2.3%, 1.4%, 2.4%, 2.2%, 1.0%, 1.0%, and 1.3%, respectively. Conclusion: The established method is accurate, reliable, and could be used for the simultaneous determination of the seven components in F. suspense, which provides a scientific basis for the quality evaluation of F. suspense.

Effect of arctigenin on apoptosis of mouse erythroleukemia FBL-3 cells and its mechanism / 中华实用儿科临床杂志

Objective To investigate the effect of arctigenin on the apoptosis of FBL-3 cells and its mechanisms.Methods The mouse erythroleukemia FBL-3 cells were taken as subjects.The untreated FBL-3 parental cells were taken as the control group,and the FBL-3 cells treated with 20 mg/L arctigenin for 24 h were taken as the experimental group.The effects of arctigenin on the apoptosis of the mouse erythroleukemia FBL-3 cells were determined by agarose gel electrophoresis and flow cytometry assay.The changes of apoptosis-related genes were analyzed by reverse transcriptase-polymerase chain reaction(RT-PCR).Results The agarose gel electrophoresis results revealed that the " DNA ladder" was displayed in the experimental group compared to the control group.The flow cytometry data demonstrated that the apoptosis number of FBL-3 cells in the experimental group was markedly increased compared to that in the control group (t =60.681,P =0.000).The RT-PCR assay results suggested that the expressions of Bcl-2 and IAP-1 gene were down-regulated in the experimental group compared to the control group (t =14.732,29.702,all P =0.000),while the expressions of Bax and Smac gene were up-regulated(t =6.721,8.499;P =0.003,0.001),but the expression of Bcl-XL did not change(t =0.209,P =0.844).Conclusions Arctigenin can induce the apoptosis of the mouse erythrolenkemia FBL-3 cells.Apoptosis of the mouse erythrolenkemia FBL-3 cells induced by arctigenin may be correlated to the down-regulation of Bcl-2 and IAP-1 and up-regulation of Bax as well as Smac.

Studies on the Calcium Antagonist Action of Arctigenin / 中草药

The calcium antagonist action of arctigenin (ACT) was studied in order to verify the ef-fect of Fructus Arctii for the relieve of exterior syndrome. Muscular contraction of isolated rat trachea,colon, pulmonary artery and thoracic aorta induced by KC1, that of guinea pig trachea induced by CaCI2,before and after the addition of ACT were assessed and their contraction-response curves drawn and PD'2calculated according to Scott. The inhibition rate of two-phase contraction of guinea pig trachea induced byacetylcholine chloride (Ach) in comparison with verapamil (VER) was calculated. Results of the studyshowed that ACT could non-compatitively antagonize the muscular contraction of the test specimens withPD'2 of 4.01, 5.11, 5.98 and 6.05 respectively. Similar to VER, ACT could non-competitively antagonizethe isolated guinea pig trachea with PD'2 of 4.04 and 5.62 respectively. Both of them could inhibit thefirst phase contraction induced by Ach with inhibition rates of 66.14% and 81.42% respectively. It wasconcluded that ACT, as the active constituent of Fructus Arctii, relaxed smooth muscle contraction byblocking the potential dependant Ca2+ channel and the internal release of Ca2+.