Study on Protective Effects of Scoparone on Acute Liver Injury Induced by CCl 4 in Mice / 中国药房

OBJECTIVE：To study the protec tive effects of scoparone on acute liver injury induced by CCl 4 in mice and its potential molecular mechanism. METHODS ：Fifty male Kunming mice were randomly divided into normal control group ，model group，silymarin group （positive control ，120 mg/kg），scoparone high-dose and low-dose groups （60，30 mg/kg），with 10 mice in each group. Administration groups were given relevant medicine intragastrically. Normal control group and model group were given constant volume of 0.5％ sodium carboxymethyl cellulose solution ，once a day ，for 7 days. Two hours after last medication ， except normal control group was intraperitoneally injected constant volume of olive oil ，other groups were intraperitoneally injected 0.1％ CCl4 olive oil solution （10 mL/kg）at one time to establish the acute liver injury model. The pathological changes of liver tissues in mice were observed by HE staining ；the activity of AST ，ALT，SOD and CAT and the contents of IL- 1β，IL-6，TNF-α and MDA in serum were measured by ELISA ；the phosphorylation of nuclear factor κB（NF-κB）pathway related proteins （NF-κB p65，IκBα）in liver tissue were detected by Western blotting assay. RESULTS ：Compared with normal control group ，serum activities of AST and ALT ，the contents of MDA ，IL-1β，IL-6 and TNF-α were significantly increased in model group，the activities of SOD and CAT were decreased significantly （P＜0.05）；obvious pathological changes were observed in liver tissues ； phosphorylation levels of NF-κB p65 and IκBα protein in liver tissues were significantly increased（P＜0.05）. Compared with model group ，the activities or contens of related factors in serum of mice were significantly reversed in silymarin group and scoparone high-dose and low-dose groups （P＜0.05）；the pathological changes of liver tissues were significantly reduced ；the phosphorylation levels of NF-κB p65 and IκBα protein in liver tissues were significantly reduced（P＜0.05）. CONCLUSIONS ： Scoparone has a protective effect on CCl 4-induced acute liver injury in mice ，which is related to reducing oxidative stress levels and blocking the activation of NF-κB pathway，thereby inhibiting inflammatory response.

A new lignan from Artemisiae Annuae Herba / 中草药

Objective: To study the constituents from the dried aboveground part of Artemisia annua. Methods: The chemical constituents were isolated by various chromatographic techniques and their structures were elucidated by spectroscopic analyses and comparison of NMR data with those reported in literatures. Results: Six compounds were obtained and characterized as artelignan (1), scopoletin (2), scoparone (3), chrysosplenol B (4), jaceidin (5) and mikanin (6). Conclusion: Compound 1 is identified as a new compound named artelignan.

Effects of scoparone on the expression of RORγt and related factors in experimental allergic rhinitis rats / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the effects of behavior, pathology, the serum IL-17, IL-23 level, and the expressing of RORγt, IL-17 and IL-23 mRNA in nasal tissues of experimental allergic rhinitis rats after the scoparone treatment. METHODS The animal model were divided into 4 groups: normal control group(group NC), allergic rhinitis group(group AR), artemolactone group(group Sco) and dexamethasone group(group Dxm). The symptom score, HE staining of the nasal mucosa, IL-17 and IL-23 level in serum measured by ELISA, the RORγt, IL-17 and IL-23 mRNA level detected by RTPCR. RESULTS Symptoms and inflammatory pathology were relieved in the experimental group after scoparone treatment. The serum levels of the IL-17, IL-23 in group Sco and group Dxm were little higher than that in group NC. The levels of RORγt, IL-17 and IL-23 mRNA in group AR were significantly higher than that in the other three groups. The levels of RORγt, IL-17 and IL-23 mRNA in group Sco and group Dxm were little higher than that in the group NC. CONCLUSION Sco could significantly inhibited or eliminated the allergy symptoms of AR in rats, and could reduce the severity and inflammatory response of diseases.

Chemical constituents from pericarpium of Metaplexis japonica / 中草药

Objective: To study the chemical constituents from the pericarpium of Metaplexis japonica. Methods: The petroleum ether, dichloromethane and ethyl acetate fractions of 95% ethanol extract from M. japonica were isolated and purified by silica gel, Sephadex LH-20, MPLC and preparative HPLC, etc. The structures of the obtained compounds were identified by physicochemical properties and spectral data. Results: Ten compounds were isolated and identified as (15β,21α)-dihydroxypregna-17,20-trimethylene oxide-4,6-dien-3-one (1), 25-hydroperoxycyc-loart-23-en-3β-ol (2), α-amyrin (3), scoparone (4), aleuritin (5), syringaresinol (6), pergularin-3-O-β-D-oleanodropyranose (7), (20β)-21-dihydroxypregna-4,6-dien-3-one (8), blumenol A (9) and 4-pregnen-20,21- diol-3-one (10). Conclusion: Compound 1 is a new C21 steroidal compound named metajapogenin F, compounds 2 and 10 are isolated from the genus Metaplexis for the first time, and compound 8 is isolated from this plant for the first time.

Solation, Purification and Identification of Antioxidant Compositions in Melilotus officinalis L / 中国药学杂志

OBJECTIVE: To study the chemical constituents from Melilotus officinalis(L.) and their anti-oxidant activities. METHODS: DPPH Assay and β-carotene bleaching assay were used for screening the fraction of anti-oxidant activity in the 70% ethanol extract of Melilotus officinalis(L.). The isolation and purification were performed by silica,Sephadex LH-20 and preparative columns. The structures of the obtained compounds were identified by physicochemical properties and spectral data. Then their anti-oxidant activities were evaluated. RESULTS: The ethyl acetate fraction from Melilotus officinalis(L.) showed the most significant anti-oxidant activity. Thirteen compounds were isolated and identified as kaempferol(1), luteolin(2), quercetin(3), hesperidin(4), rutin(5), apigenin(6), salicylic acid(7), coumarin(8), scoparone(9), 7-methoxycoumarin(10), fumaric acid(11), caffeic acid(12), and p-hydroxybenzoate-4-O-α-D-mannopyranosyl-(1→3)-α-L-rhamnoside(13).Compounds 2, 4, 7 and 12 exhibited strong scavenging activities against DPPH free radical with the IC50 values of 25.89, 14.09, 28.03, and 23.04 μg•mL-1, respectively; compounds 4, 7, 12 and 13 exhibited strong scavenging activities against ABTS free radical with the IC50 values of 10.48, 23.77, 22.79, and 25.20 μg•mL-1, respectively. CONCLUSION: Compounds 3, 7, 9, 10, 11, 12, and 13 are isolated from this plant for the first time. Compound 13 is a new compound not reported in the literature; compounds 2, 4, 7, and 12 have strong ability to scavenge DPPH free radicals; compounds 4, 7, 12, and 13 have strong ability to scavenge ABTS free radicals.

Simultaneous Determination of Four Compounds from Artemisia capillaris using High Performance Liquid Chromatography-Ultraviolet Detector (HPLC-UVD) and Their Quantitative Study in Artemisia Genus

Artemisia capillaris Thunb. (Compositae) is a native herb of East Asian countries and has used for the treatment of jaundice, high liver fever, and digestive diseases for a long time, as well as being developed as the source of herbal preparations until now. The major components from A. capillaris were chlorogenic acid (1) and its derivatives substituted with caffeoyl moieties, such as 3,5-dicaffeoylquinic acid (2) and 4,5-dicaffeoylquinic acid (3), and coumarins, such as scoparone. In the study, four compounds, chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and scoparone (4) in the 70% ethanolic extract of A. capillaris were simultaneously determined by using HPLC-UVD system. This method was validated with the terms of linearity, precious and accuracy according to ICH guidelines. The developed method was successfully applied for the quantitative analysis of Artemisia genus, A. capillaris, A. iwayomogi, A. princeps, and A. argyi, distributed in Korea.

Simultaneous Determination of Six Components in Ganyan Kangfu Pills by HPLC / 中国药师

Objective:To establish a wavelength switching HPLC with gradient elution for the simultaneous determination of six active components, [(R,S)-goitrin, salvianolic acid B, tanshinone IIA, luteolin-7-glucoside, 4-hydroxyacetophenone and scoparone ] in Ganyan Kangfu pills .Methods:An Agilent TC-C18 (250 mm ×4.6 mm, 5 μm) chromatographic column was used with the mobile phase consisting of acetonitrile and 0.1％ phosphoric acid solution with gradient elution .The flow rate was 0.9 ml· min-1 , and the column temperature was 25 ℃.( R,S)-Goitrin was detected at 245 nm, salvianolic acid B and tanshinone IIA were detected at 270 nm, luteolin-7-glucoside was detected at 348 nm, 4-hydroxyacetophenone and scoparone were detected at 278 nm.The sample volume was 10 μl.Results:The linear range of (R,S)-goitrin, salvianolic acid B, tanshinone ⅡA, luteolin-7-glucoside, 4-hydroxyacetophe-none and scoparone was 1.99-49.75 μg· ml-1(r=0.9999), 18.66-466.50 μg· ml-1(r=0.9994), 2.25-56.25 μg· ml-1(r=0.9998), 2.62-65.50 μg· ml-1(r=0.9998), 2.48-62.00 μg· ml-1(r=0.9992) and 2.55-63.75μg· ml-1(r=0.9996), respectively.The average recovery was 98.42％(RSD=0.83％), 99.56％(RSD=1.04％), 97.96％(RSD=1.53％), 96.84％(RSD=0.78％), 98.10％(RSD=1.44％) and 97.82％(RSD=1.34％)(n=6), respectively.Conclusion: The method with good reproducibility is simple and accurate , which provides a new way for the quality evaluation of Ganyan Kangfu pills .

Antifungal activity against colletotrichum acutatum and colletotrichum gloeosporioides of the major constituents from wood sawdust of platymiscium gracile benth / Actividad antifúngica contra colletotrichum acutatum y colletotrichum gloeosporioides de los constituyentes mayoritarios del aserrín de madera de platymiscium gracile benth

The tree tomato (Solanum betaceum Cav., Solanaceae) anthracnose, caused by the fungi Colletotrichum acutatum and Colletotrichum gloeosporioides, is the most important disease of this crop in Colombia for its wide distribution and the losses it causes. In the present work, the in vitro antifungal activity of the soluble fractions in n-hexane, dichloromethane, and ethyl acetate, and their major constituents from the sawdust of timber specie Platymiscium gracile Benth. (Fabaceae) against both fungi was evaluated. The n-hexane-soluble fraction exhibited the greatest inhibitory effect. The metabolites homopterocarpin (a pterocarpan, 0.39 percent dry weight), calycosin (an isoflavone, 2.01 percent) and scoparone (a coumarin, 1.48 percent) were isolated for the first time from wood sawdust of P. gracile. The structure of these compounds was determined by 1H and 13C NMR analyses. The three compounds tested showed significant antifungal activity.

La antracnosis del tomate de árbol (Solanum betaceum Cav., Solanaceae), ocasionada por los hongos Colletotrichum acutatum y Colletotrichum gloeosporioides, es la enfermedad más importante de este cultivo en Colombia por su amplia distribución y las pérdidas que ocasiona. En el presente trabajo se evaluó la actividad antifúngica in vitro de las fracciones solubles en n-hexano, diclorometano y acetato de etilo, y sus componentes mayoritarios, del aserrín de la especie maderable Platymiscium gracile Benth. (Fabaceae), contra ambos hongos. La fracción en n-hexano exhibió el mayor efecto inhibitorio. Los metabolitos homopterocarpina (un pterocarpano; 0.39 por ciento del peso seco de aserrín), calicosin (una isoflavona; 2.01 por ciento) y escoparona (una cumarina; 1.48 por ciento) se aislaron por primera vez desde el aserrín de madera de P. gracile empleando técnicas cromatográficas. La estructura de los compuestos se determinó por análisis de RMN de 1H y 13C. Los tres metabolitos mostraron una actividad antifúngica significativa contra ambos hongos.

Chemical Constituents from Pericarpium Citri Reticulatae / 中草药·英文版

Objective To study the chemical constituents from Pericarpium Citri Reticulatae (Citrus reticulata). Methods The chemical constituents were isolated and purified by silica gel column, Sephadex LH-20, and ODS column chromatography. The structures were identified by spectral data. Results Nineteen compounds were isolated and identified as 4′,5,6,7-tetramethoxyflavone (1), 3,3′,4′,5,6,7,8-heptmethoxyflavone (2), sinensetin (3), 5-O-demethylnobiletin (4), tangeretin (5), nobiletin (6), apigenin (7), 5-O-desmethyltangeretin (8), 5,7-dihydroxy-3,3′,4′,6-tetramethoxyflavone (9), pachypodol (10), 4′,5,6,7-tetramethoxyflavanone (11), 3′,4′,5,7,8-pentamethoxyflavanone (12), agestricin C (13), scoparone (14), isoscopoletin (15), hesperidin (16), didymin (17), methylhesperidine (18), and naringin (19). Conclusion Compounds 9-15 are obtained from this plant for the first time.

Investigation of three coumarin components in thorns of Gleditsia sinensis and their cytotoxic activity to tumor cells / 中草药

Objective: To investigate the chemical constituents from Gleditsiae Spina (the thorns of Gleditsia sinensis). Methods: The compounds were isolated and purified by silica gel, Sephadex LH-20 column chromatographic techniques, and their chemical structures were confirmed on the basis of spectroscopic analysis by the physicochemical properties. Cytotoxic activity using the MTT colorimetry method was performed to measure the inhibitory effect of the compounds on cell proliferation of HepG2, A-549, and EC109. Results: Three coumarins were obtained from the ethyl acetate soluble fraction of the 90% ethanolic extract and their structures were identified as scoparone (1), isoscopoletin (2), and 6-amino-7-methoxycoumarin (3). Conclusion: Compound 3 is a novel coumarin named gledisinmarin A. Compounds 1 and 2 are isolated from this plant for the first time. Compound 2 displays the stronger cytotoxicity against A549 cell with an IC50 value of 34.47 μg/mL, while cisplatin with an IC50 value of 11.50 μg/mL as a positive control.

Chemical constituents and bioactivities of Zanthoxylum avicennae / 中草药

Objective: To study the chemical constituents of the ethyl acetate (EtOAc) extract from Zanthoxylum avicennae and their bioactivities. Methods: The compounds were isolated by Diaion HP-20, Toyopearl HW-40, MCI-Gel CHP-20, Sephadex LH-20, RP18, and silica gel column chromatographic methods. Their structures were identified on the basis of physicochemical and spectroscopic analyses; The antibacterial activity and toxicity of compounds 1-3 were investigated by the lethal-to-prawn larva bioactivity experiment and the slanting test-tube experiment. Results: Nine constituents were isolated from the EtOAc and n-BuOH fractions of Z. avicennae. Their structures were identified as dihydrokaempferide (1), scoparone (2), 1H-indole-3-carboxylic acid (3), 6-oxo-2(4-hydroxy-3,5-dimethoxyphenyl)-3,7-dioxabicyclo-[3.3.0]-octane (4), suberic acid (5), 4,4-dinethyl-1,7-heptanedioic acid (6), methyl syringate (7), β-sitosterol (8), and preskimmianine (9). Compound 2 displayed more stronger antifungal activity against Exserohilum turcicum Pass and Fusarium graminearum Schwabe, with the MIC value of 15.63 μg/mL. Compounds 1-3 had shown more stronger toxicity towards brine shrimp, with LC50 values of 9.8, 2.6, and 4.9 μg/mL, respectively. Conclusion: All the compounds are isolated from Z. avicennae for the first time except preskimmianine. Compound 2 displays more stronger antifungal activity, while compounds 1-3 have shown more stronger cytotoxicity towards brine shrimp.

Chemical constituents from aerial parts of Artemisia dracunculus var. turkestanica / 中草药

Objective: To study the chemical constituents from the aerial parts of Artemisia dracunculus var. turkestanica. Methods: The compounds were isolated and purified by silica gel, RP18, and Sephadex LH-20 column chromatography, and their structures were elucidated on the basis of spectral data and physicochemical properties. Results: Nineteen compounds were obtained from ethyl acetate fraction of the 95% ethanol extract from the aerial parts of A. dracunculus var. turkestanica and identified as 1-monopalmitin (1), α-methyl linolenate (2), 1-monoolein (3), 3-O-acetyl ursolic acid (4), 3β, 22β, 24-trihydroxy-olean-12-en (5), oleic acid (6), palmitic acid (7), 1-monolinolein (8), vanillic acid (9), hesperetin (10), naringenin (11), quercetin (12), rutin (13), scoparone (14), scopoletin (15), 7-hydroxycoumarin (16), daphnetin (17), caffeic acid (18), and chlorogenic acid (19). Conclusion: All the compounds are isolated from A. dracunculus var. turkestanica for the first time. Compounds 1-5 and 8 are firstly reported in the plants of Artemisia L.

Leishmanicidal and cholinesterase inhibiting activities of phenolic compounds of Dimorphandra gardneriana and Platymiscium floribundum, native plants from Caatinga biome / Atividade inibitória Leishmanicida e Colinesterásica de Compostos Fenólicos de Dimorphandra gardneriana e Platymiscium fliribunbum, plantas nativas do bioma Caatinga

In recent years, the Brazilian Health Ministry and the World Health Organization have supported research into new technologies that may contribute to the surveillance, new treatments, and control of visceral leishmaniasis within the country. In light of this, the aim of this study was to isolate compounds from plants of the Caatinga biome, and to investigate their toxicity against promastigote and amastigote forms of Leishmania infantum chagasi, the main responsible parasite for South American visceral leishmaniasis, and evaluate their ability to inhibit acetylcholinesterase enzyme (AChE). A screen assay using luciferase-expressing promastigote form and an in situ ELISA assay were used to measure the viability of promastigote and amastigote forms, respectively, after exposure to these substances. The MTT colorimetric assay was performed to determine the toxicity of these compounds in murine monocytic RAW 264.7 cell line. All compounds were tested in vitro for their anti-cholinesterase properties. A coumarin, scoparone, was isolated from Platymiscium floribundum stems, and the flavonoids rutin and quercetin were isolated from Dimorphandra gardneriana beans. These compounds were purified using silica gel column chromatography, eluted with organic solvents in mixtures of increasing polarity, and identified by spectral analysis. In the leishmanicidal assays, the compounds showed dose-dependent efficacy against the extracellular promastigote forms, with an EC50 for scoporone of 21.4µg/mL, quercetin and rutin 26 and 30.3µg/mL, respectively. The flavonoids presented comparable results to the positive control drug, amphotericin B, against the amastigote forms with EC50 for quercetin and rutin of 10.6 and 43.3µg/mL, respectively. All compounds inhibited AChE with inhibition zones varying from 0.8 to 0.6, indicating a possible mechanism of action for leishmacicidal activity.

Nos últimos anos, o Ministério da Saúde do Brasil e a Organização Mundial da Saúde tem apoiado a investigação de novas tecnologias que possam contribuir para a vigilância, novos tratamentos e controle da leishmaniose visceral no país. Assim, o objetivo deste trabalho foi isolar compostos de plantas do bioma Caatinga, e investigar a toxicidade destes compostos contra as formas promastigotas e amastigotas de Leishmania infantum chagasi, principal parasita responsável pela leishmaniose visceral na América do Sul, e avaliar a sua capacidade para inibir a enzima acetil-colinesterase (AChE). Após a exposição aos compostos em estudo, foram realizados testes utilizando a forma promastigota que expressa luciferase e ELISA in situ para medir a viabilidade das formas promastigotas e amastigota, respectivamente. O ensaio colorimétrico MTT foi realizado para determinar a toxicidade destas substâncias utilizando células monocíticas murina RAW 264.7. Todos os compostos foram testados in vitro para as sua propriedade anti-colinesterásica. Um cumarina, escoparona, foi isolada a partir de hastes de Platymiscium floribundum, e os flavonóides, rutina e quercetina, foram isolados a partir de grãos de Dimorphandra gardneriana. Estes compostos foram purificados, utilizando cromatografia em coluna gel eluída com solventes orgânicos em misturas de polaridade crescente, e identificados por análise espectral. Nos ensaios leishmanicidas, os compostos fenólicos mostraram eficácia contra as formas extracelulares promastigotas, com EC50 para escoporona de 21.4µg/mL e para quercetina e rutina 26 e 30.3µg/mL, respectivamente. Os flavonóides apresentaram resultados comparáveis à droga controle, a anfotericina B, contra as formas amastigotas com EC50 para quercetina e rutina de 10.6 e 43.3µg/mL, respectivamente. Os compostos inibiram a enzima AChE com halos de inibição variando de 0,8 a 0,6cm, indicando um possível mecanismo de ação para a atividade leishmanicida.

Enrichment process of scoparone from Yinchenhao Decoction with macroporous resin / 中草药

Object To study the optimal conditions and parameters for the enrichment process of scoparone in Yinchenhao Decoction (YCHD) with macroporous resin. Methods With the enrichment degree of scoparone in YCHD as an index, the optimal conditions of the enrichment process were investigated. Results The optimal conditions were that the aqueous extract of YCHD was taken into the macroporous resin column, and kept for 30 min, then the resin was washed by five times of distilled water to get rid of impurity and three — four times of 70% ethanol to elute scoparone. Conclusion The process is feasible to enrich effective ingredient scoparone from YCHD.

The effects of scoparone on the content of IL-4 and IFN-r in asthmatic guinea pigs / 中国药理学通报

AIM To study the changes of IgE, IL-4 and IFN-y in serum and pulmonary tissue homogenate of asthmatic guinea pigs and the effects of scoparone on them. METHODS To divide animals into three groups: control, asthma and scoparone treatment groups. Choose the model guinea pigs of asthma sensitized with OA, and observe the changes of IgE, IL-4 and IFN-r in serum and pulmonary tissue homogenate of asthmatic guinea pigs and the effects of scoparone on them by means of chemolumi nescence, radio immunoassay, enzyme-linked immunoabsordent assay. RESULTS IgE and IL-4 in serum and pulmonary tissue homoge-nate of asthmatic guinea pigs obviously increase (P