Three new anthraquinones from Prismatomeris tetrandra (Roxb.) K. Schum and their protective effects in neuroblastoma cells / 药学学报

Three new anthraquinones were isolated from the 80% ethanol extract of Prismatomeris tetrandra by silica gel, MCI, ODS column chromatography and high performance preparative liquid chromatography (HPLC). The structures of the new compounds were identified by mass spectrometry, nuclear magnetic resonance and other spectroscopic methods as 6-hydroxy-1,2,3-trimethoxy-7-methylanthracene-9,10-dione (1), 6-(hydroxymethyl)-1,2,3-trimethoxyanthracene-9,10-dione (2) and 7-hydroxy-6-(hydroxymethyl)-1,2-dimethoxyanthracene-9,10-dione (3). Compounds 1, 2 and 3 showed protective effects against monosodium glutamate-induced damage in SH-SY5Y neuroblastoma cells, with the cell survival rates elevated 18.45%, 4.31%, and 7.65%, respectively.

Rubiadin exerts an acute and chronic anti-inflammatory effect in rodents / A Rubiadina exerce um efeito anti-inflamatório agudo e crônico em roedores

Rubiadin is identified as a bioactive anthraquinone that exists in some quinone rich plants. The current research was carried out to evaluate the potential anti-inflammatory impact of Rubiadin in acute and chronic inflammation test models in rodents. The anti-inflammatory activity of Rubiadin was examined in cotton pellet-induced granuloma and carrageenan-induced edema as chronic and acute inflammation models in rats. TNF-α level and histopathological changes were assessed using sampled foot tissue of rat in the acute model. Also, the IL-1β level was assessed in the chronic model. One-way ANOVA (post hoc Tukey’s) analysis was used for comparing the groups. Rubiadin (0.5 mg/kg, i.p.) induced a significant reduction in TNF α level and the paw edema compared to the control group in carrageenan test. Also, it was observed that the anti-inflammatory activity of Rubiadin (0.5 mg/kg, i.p.) is comparable to mefenamic acid (30 mg/kg, i.p.) as the standard drug. Rubiadin was effective in granuloma induced by cotton pellet concerning the granuloma and transudate formation amount. Rubiadin's anti-inflammatory effects were associated with a significant IL-1β decrease in this model. The results suggest that Rubiadin as a natural compound can possess significant peripheral anti-inflammatory impacts.

A rubiadina é identificada como uma antraquinona bioativa que existe em algumas plantas ricas em quinonas. A presente pesquisa foi realizada para avaliar o potencial impacto anti-inflamatório da rubiadina em modelos de teste de inflamação aguda e crônica em roedores. A atividade anti-inflamatória da rubiadina foi examinada em granuloma induzido por pellet de algodão e edema induzido por carragenina como modelos de inflamação crônica e aguda em ratos. O nível de TNF-α e as alterações histopatológicas foram avaliados usando amostra de tecido do pé de rato no modelo agudo. Além disso, o nível de IL-1β foi avaliado no modelo crônico. A análise ANOVA de uma via (post hoc de Tukey) foi usada para comparar os grupos. A rubiadina (0,5 mg / kg, i.p.) induziu uma redução significativa no nível de TNF α e no edema da pata em comparação com o grupo de controle no teste de carragenina. Além disso, foi observado que a atividade anti-inflamatória da rubiadina (0,5 mg / kg, i.p.) é comparável ao ácido mefenâmico (30 mg/kg, i.p.) como o fármaco padrão. A rubiadina foi eficaz no granuloma induzido por pellet de algodão no que diz respeito à quantidade de granuloma e formação de transudato. Os efeitos anti-inflamatórios da rubiadina foram associados a uma redução significativa de IL-1β nesse modelo. Os resultados sugerem que a rubiadina como um composto natural pode ter impactos anti-inflamatórios periféricos significativos.

Rubiadin exerts an acute and chronic anti-inflammatory effect in rodents

Abstract: Rubiadin is identified as a bioactive anthraquinone that exists in some quinone rich plants. The current research was carried out to evaluate the potential anti-inflammatory impact of Rubiadin in acute and chronic inflammation test models in rodents. The anti-inflammatory activity of Rubiadin was examined in cotton pellet-induced granuloma and carrageenan-induced edema as chronic and acute inflammation models in rats. TNF- level and histopathological changes were assessed using sampled foot tissue of rat in the acute model. Also, the IL-1 level was assessed in the chronic model. One-way ANOVA (post hoc Tukeys) analysis was used for comparing the groups. Rubiadin (0.5 mg/kg, i.p.) induced a significant reduction in TNF level and the paw edema compared to the control group in carrageenan test. Also, it was observed that the anti-inflammatory activity of Rubiadin (0.5 mg/kg, i.p.) is comparable to mefenamic acid (30 mg/kg, i.p.) as the standard drug. Rubiadin was effective in granuloma induced by cotton pellet concerning the granuloma and transudate formation amount. Rubiadins anti-inflammatory effects were associated with a significant IL-1 decrease in this model. The results suggest that Rubiadin as a natural compound can possess significant peripheral anti-inflammatory impacts.

Resumo A rubiadina é identificada como uma antraquinona bioativa que existe em algumas plantas ricas em quinonas. A presente pesquisa foi realizada para avaliar o potencial impacto anti-inflamatório da rubiadina em modelos de teste de inflamação aguda e crônica em roedores. A atividade anti-inflamatória da rubiadina foi examinada em granuloma induzido por pellet de algodão e edema induzido por carragenina como modelos de inflamação crônica e aguda em ratos. O nível de TNF- e as alterações histopatológicas foram avaliados usando amostra de tecido do pé de rato no modelo agudo. Além disso, o nível de IL-1 foi avaliado no modelo crônico. A análise ANOVA de uma via (post hoc de Tukey) foi usada para comparar os grupos. A rubiadina (0,5 mg / kg, i.p.) induziu uma redução significativa no nível de TNF e no edema da pata em comparação com o grupo de controle no teste de carragenina. Além disso, foi observado que a atividade anti-inflamatória da rubiadina (0,5 mg / kg, i.p.) é comparável ao ácido mefenâmico (30 mg/kg, i.p.) como o fármaco padrão. A rubiadina foi eficaz no granuloma induzido por pellet de algodão no que diz respeito à quantidade de granuloma e formação de transudato. Os efeitos anti-inflamatórios da rubiadina foram associados a uma redução significativa de IL-1 nesse modelo. Os resultados sugerem que a rubiadina como um composto natural pode ter impactos anti-inflamatórios periféricos significativos.

Spectrum-effect relationship of total anthraquinone extract of Cassia seeds against fluorouracil-induced liver injury in mice / 南方医科大学学报

OBJECTIVE@#To investigate the spectrum-effect relationship between the total anthraquinone extract of Cassia seeds and fluorouracil (5-Fu)-induced liver injury in mice and identify the effective components in the extract.@*METHODS@#A mouse model of liver injury was established by intraperitoneal injection of 5-Fu, with bifendate as the positive control. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and myeloperoxidase (MPO), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in the liver tissue were detected to investigate the effect of the total anthraquinone extract of Cassia seeds (0.4, 0.8 and 1.6 g/kg) on liver injury induced by 5-Fu. HPLC fingerprints of 10 batches of the total anthraquinone extracts were established to analyze the spectrum- effectiveness of the extract against 5- Fu- induced liver injury in mice and screen the effective components using the grey correlation method.@*RESULTS@#The 5- Fu- treated mice showed significant differences in liver function parameters from the normal control mice (P < 0.05), suggesting successful modelling. Compared with those in the model group, serum ALT and AST activities were decreased, SOD and T- AOC activities significantly increased, and MPO level was significantly lowered in the mice treated with the total anthraquinone extract (all P < 0.05). HPLC fingerprints of the 31 components in the total anthraquinone extract of Cassia seeds showed good correlations with the potency index of 5-Fu-induced liver injury but with varying correlation strengths. The top 15 components with known correlations included aurantio-obtusina (peak 6), rhein (peak 11), emodin (peak 22), chrysophanol (peak 29) and physcion (peak 30).@*CONCLUSION@#The effective components in the total anthraquinone extract of Cassia seeds, including aurantio-obtusina, rhein, emodin, chrysophanol, and physcion, are coordinated to produce protective effects against 5-Fu-induced liver injury in mice.

Long-chain alkanol-alkyl carboxylic acid-based low-viscosity hydrophobic deep eutectic solvents for one-pot extraction of anthraquinones from Rhei Radix et Rhizoma / 药物分析学报

Natural long-chain alkanol and alkyl carboxylic acid were used to prepare novel hydrophobic deep eutectic solvents(HDESs).These HDESs are liquid at room temperature and have low viscosity(＜12.26 mPa-s),low polarity(lower than that of methanol,ChCl-based deep eutectic solvents and other reported HDESs),and low density(＜0.928 g/mL).A simple one-pot method based on a novel HDES-water two-phase extraction system was constructed for the extraction of weak-polarity bioactive components,anthraquinones,from Rhei Radix et Rhizoma.This HDES-based new extraction method does not consume hazardous organic solvents and can obtain a total anthraquinone yield of 21.52 mg/g,which is close to that obtained by the Chinese pharmacopoeia method(21.22 mg/g)and considerably higher than those by other reported HDESs-based extraction methods(14.20-20.09 mg/g,p＜0.01).The high extraction yield can be mainly attributed to the severe destruction of the RRR cell walls by the extraction system and the excellent dissolving ability of novel HDESs for anthraquinones.

Study on potential hepatotoxicity of main monomers of Polygonum multiflorum based on liver micro-tissue / 中国中药杂志

In this study, we aimed to establish a rat liver micro-tissue evaluation system to evaluate the hepatotoxicity of the main monomers in Polygonum multiflorum. Rat primary hepatocytes were isolated and purified by two-step in situ perfusion method to prepare hepatic parenchymal cells. The ultra-low adsorption plate and the inverted model were used to establish an in vitro hepatotoxicity evaluation system. After the system was established, the main monomer components(monanthone with emodin type, rhein, emodin, emodin-8-O-β-D-glucopyranoside, physcion) of P. multiflorum were selected for in vitro hepatotoxicity evaluation. This study showed that the primary cells of the liver can form liver micro-tissues in the low adsorption plate method and the mold perfusion method, with good liver structure and function, which can be used to evaluate the hepatotoxicity of the drug to be tested after long-term administration. The five monomers to be tested in P. multiflorum can significantly affect the proliferation of primary liver micro-tissues in rats in a dose-and time-dependent manner. The hepatotoxic effects were as follows: monanthone with emodin type > rhein > emodin > emodin-8-O-β-D-glucopyranoside > physcion. The results suggested that the emodin-type monoterpene and rhein might be the potential hepatotoxic components, while the metabolites of emodin-8-O-β-D-glucoside and emodin methyl ether showed more toxic risks. The rat primary hepatocyte micro-tissue model system established in this experiment could be used to achieve long-term drug administration in vitro, which was consistent with the clinical features of liver injury caused by long-term use of P. multiflorum. The experimental results provided important information and reference on the clinical application and toxic component of P. multiflorum.

Effects of different decoction methods on functional components of rhubarb based on pseudo-targeted metabolomics / 中草药

Objective: The effects of different decoction time and different decoction temperature on its chemical constituents of anthraquinones, anthrone, and tannins were revealed by pseudo-targeted metabolomics, which provided the basis for clinical use of rhubarb. Methods: Using LTQ-Orbitrap-MSn to accurately excavate the characteristic ions of various chemical components of rhubarb, the ion pairs of chemical components were determined as much as possible by characteristic ions to obtain the peak area of various components by using the MRM mode of QQQ-MS. The effects of different decoction methods on the chemical components of rhubarb was compared by multivariate statistical analysis combined with paired t test. Results: Both decoction time and decoction temperature have impacts on the chemical components of rhubarb decoction. Short-term decocting for 15 min was beneficial to the dissolution of dianthrone glycosides and anthraquinone glycosides, while long-term decocting for 60 min was beneficial to the dissolution of tannins; Compared with boiling water maceration, boiling water decoction was more favorable for the dissolution of anthraquinones and tannins. Conclusion: This paper adopts the method of pseudo-targeted metabolomics, and clearly points out that short-term decocting or maceration is conducive to exerting the laxative effect of rhubarb, and long-time boiling decocting is conducive to exerting clearing heat and detoxifying efficacy, which provides a reference for the clinical application of rhubarb.

A systematic evaluation of biopharmaceutics classification system of main efficacy and toxic ingredients in Polygoni Multiflori Radix Praeparata / 中草药

Objective: Biopharmaceutics classification system (BCS) of five main pharmacological/toxic components (gallic acid, emodin, stilbene glycoside, physcion, and emodin-8-O-β-D-glucoside) of Polygoni Multiflori Radix Praeparata (PMRP) were carried out. Methods: The solubility and permeability of each representative component were studied by equilibrium solubility method and everted intestinal sac method, respectively. Using two softwares (Pipeline Pilot 7.5, ChemDraw 7.0) to predict the solubility and permeability parameters of each component. Classical BCS classification of measured and predicted values of representative components was conducted according to Food and Drug Administration (FDA) standards, and their correlation was evaluated. Results: The emodin, emodin-8-O-β-D-glucoside, and physcion in PMRP was preliminary determined as BCS IV drugs. THSG and gallic acid belong to BCS III drugs, and permeability was the main limiting factor in their absorption process. There was software which predicts false positives of anthraquinone in BCS classification studies. Conclusion: In this study, five main pharmacodynamic/toxic components of PMRP were classified by BCS method, which provided data support and technical reference for in vivo absorption prediction and in vitro safety evaluation of traditional Chinese medicine.

Influences of ancient processing method steaming with black bean and drying and pharmacopoeia processing method continuous steaming with black bean decoction on 12 components of Polygoni Multiflori Radix / 中草药

Objective: To explore the influence of ancient processing method black bean "nine cycles of steaming and drying" and modern pharmacopoeia processing method continuous steaming with black bean decoction on the main components of Polygoni Multiflori Radix (PMR). Methods: Simulating the time arrangement of "nine cycles of steaming and drying", samples were prepared using three processing methods: ancient method that raw PMR (rPMR) and black bean were steamed in layers and then dried, modern method that rPMR were steamed with black bean decoction and then dried, the method recorded in Chinese Pharmacopoeia that rPMR were steamed continuously with black bean decoction; The determination method of 12 components in rPMR and processed PMR (pPMR) was established using ultra-performance liquid chromatography with triple quadrupole mass spectrometry (UPLC-QqQ-MS/ MS), and 12 components in all samples processed by different methods were determined; The results was analyzed combining with t test, principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). Results: A reliable UPLC-QqQ-MS/MS method was established for the determination of emodin, physcion, rhein, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, cis-2,3,5,4'-tetrahydroxylstilbene-2-O-β-D-glucoside (cis-THSG), trans-2, 3,5,4'-tetrahydroxylstilbene- 2-O-β-D-glucoside (trans-THSG), polydatin, resveratrol, epicatechin, rutin and hyperoside. With the prolongation of steaming time, the content of 12 effective components changed obviously: the content of free anthraquinone was decreased first and then increased; The content of anthraquinone glycoside, cis-THSG, polydatin and hyperoside was increased first and then decreased; The content of trans-THSG, resveratrol, epicatechin and rutin was decreased; The components content were closely related to the auxiliary materials, steaming operation methods and processing time, the influence of operation methods was greater than that of auxiliary materials on the quality of pPMR. Conclusion: The ancient processing method steaming with black bean and drying could not be equated with the modern pharmacopoeia processing method continuous steaming with black bean decoction in terms of the content of effective components. The result provides experimental basis for inheriting and developing the traditional processing method of PMR.

Effects of pH, Temperature and Agitation on the Decolourisation of Dyes by Laccase-Containing Enzyme Preparation from Pleurotus sajor-caju

(1) Background: In this study, the effects of different pH values ​​(2.4, 3.2, 4.4 and 5.0), temperatures (30, 35, 40, 45 and 50°C) and agitation (100 rpm) on the enzymatic decolourisation of twenty-two dyes belonging to the chromophore groups anthraquinone, azo and triphenylmethane were assessed. (2) Methods: In all conditions, it was used a crude enzyme broth containing 30 U mL-1 laccases produced by Pleurotus sajor-caju PS-2001 in submerged process. (3) Results: Regarding the effects of pH values, the best results were obtained at pH 3.2 and 30°C, in which bleaching was observed for all dyes evaluated. In assays conducted at different temperatures, highest levels of decolourisation were observed at 35°C and pH 3.2 for nineteen of the dyes assessed. Thirteen dyes presented colour reduction exceeding 50% after the enzymatic treatment, including all acid and all disperse dyes evaluated. The reciprocal agitation of 100 rpm promoted negative effect on decolourisation. (4) Conclusion: From the results achieved, one can conclude that the laccase-containing preparation of P. sajor-caju PS-2001 has potential for the decolourisation of some dyes widely used in different industrial sectors, especially in the textile industry, and therefore could be used in future strategies for the biotreatment of coloured wastes.

Chemical constituents of alkaloids part of Cephalotaxus hainanensis / 中草药

Objective To study the alkaloids constituents from Cephalotaxus hainanensis. Methods The column chromatography Sephadex LH-20 and ODS were used to isolate and purify the compounds of alkaloids part of C. hainanensis. The chemical structures were identified on basis of physicochemical properties and spectral data. Results Thirteen compounds were isolated and identified as cephalotaxine (1), isocephalotaxinone (2), cephalotaxinone (3), cephalezomine G (4), (2R,3S,4S,5S)-2,3-dihydroxycephalotaxane (5), acetylcephalotaxine (6), deoxyharringtonine (7), (R)-fortunine (8), (S)-fortunine (9), isomer of 3-epi-schellhammericine (10), 1,4 (PrINH)2-anthraquinone (11), 2-ethyl-n-hexyl benzoate (12), and β-sitosterol (13). Conclusion Compounds 3-6 and 10-13 are isolated from this plant for the first time.

Bacterial degradation of anthraquinone dyes / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Anthraquinone dyes, which contain anthraquinone chromophore groups, are the second largest class of dyes after azo dyes and are used extensively in textile industries. The majority of these dyes are resistant to degradation because of their complex and stable structures; consequently, a large number of anthraquinone dyes find their way into the environment causing serious pollution. At present, the microbiological approach to treating printing and dyeing wastewater is considered to be an economical and feasible method, and reports regarding the bacterial degradation of anthraquinone dyes are increasing. This paper reviews the classification and structures of anthraquinone dyes, summarizes the types of degradative bacteria, and explores the possible mechanisms and influencing factors of bacterial anthraquinone dye degradation. Present research progress and existing problems are further discussed. Finally, future research directions and key points are presented.

Bacterial degradation of anthraquinone dyes / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Anthraquinone dyes, which contain anthraquinone chromophore groups, are the second largest class of dyes after azo dyes and are used extensively in textile industries. The majority of these dyes are resistant to degradation because of their complex and stable structures; consequently, a large number of anthraquinone dyes find their way into the environment causing serious pollution. At present, the microbiological approach to treating printing and dyeing wastewater is considered to be an economical and feasible method, and reports regarding the bacterial degradation of anthraquinone dyes are increasing. This paper reviews the classification and structures of anthraquinone dyes, summarizes the types of degradative bacteria, and explores the possible mechanisms and influencing factors of bacterial anthraquinone dye degradation. Present research progress and existing problems are further discussed. Finally, future research directions and key points are presented.

Potential of natural repellents methylanthranilate and anthraquinone applied on maize seeds and seedlings against house sparrow (Passer domesticus) in captivity / Potencial de repelentes naturais, metilantranilato e antraquinona aplicados em sementes de milho e mudas contra o pardal (Passer domesticus) em cativeiro

Abstract Various bird pests caused severe economic losses to valuable crops and fruit orchards all over the world. Among the birds, house sparrow is also considered to cause heavy plunder, not only to seeds of crops but also seedlings especially in organic farming. In present study two bird repellents, methylanthranilate and anthraquinone tested against house sparrows on maize seeds and seedlings in aviary conditions. Trial group in aviary-I, the treated maize seeds and seedlings with different doses of both bird repellents, control group in aviary-II, untreated seeds and seedlings were provided for three hours in the early morning. In each aviary, two closed circuit cameras were also installed to monitor the behavioral responses against different concentrations of both chemical repellents. Statistical analysis showed that there existed highly significant (P<0.01) variations among the trial and control groups for seeds and seedlings. By comparing both repellents, significant (P<0.05) differences were detected and anthraquinone showed better efficacy when compared to methylanthranilate, but in maize seedlings both repellents equal repellent properties. Non-significant (P>0.05) differences were observed in different grading of both natural chemical repellents for maize seeds while significant (P<0.05) variations were noticed for maize seedlings when provided to sparrows. By videotaped behavior sparrows presented manifest head juddering and feather upsetting activities by consumption of treated seeds and seedlings with higher concentrations of both natural bird repellents.

Resumo Várias pragas de aves causaram graves perdas econômicas para cultivos valiosos e pomares de frutas em todo o mundo. Entre os pássaros, o pardal da casa também é considerado um grande saqueo, não só para as sementes das culturas, mas também para as mudas, especialmente na agricultura orgânica. No presente estudo, dois repelentes de aves, metilantranilato e antraquinona testados contra pardais de casa em sementes de milho e mudas em condições de aviário. O grupo de ensaio em aviary-I, as sementes de milho tratadas e as mudas com diferentes doses de repelentes de aves, grupo de controle em aviary-II, sementes não tratadas e mudas foram fornecidas por três horas no início da manhã. Em cada aviário, duas câmeras de circuito fechado também foram instaladas para monitorar as respostas comportamentais contra diferentes concentrações de ambos os repelentes químicos. A análise estatística mostrou que existiam variações altamente significativas (P<0,01) entre os grupos de teste e controle para sementes e mudas. Ao comparar os dois repelentes, detectaram-se diferenças significativas (P<0,05) e a antraquinona apresentou maior eficácia quando comparada ao metilantranilato, mas em mudas de milho, ambos os repelentes são iguais às propriedades repelentes. As diferenças não significantes (P>0,05) foram observadas em diferentes classificações de repelentes químicos naturais para sementes de milho, enquanto as variações significativas (P<0,05) foram observadas para as mudas de milho quando fornecidas aos pardais. Por um comportamento gravado em video, os pardais apresentaram manifestações de cabeça e vibrações de penas por consumo de sementes tratadas e mudas com maiores concentrações de repelentes de aves naturais.

Decolorization of Cibacron Blue 3G-A Dye by Agaricus bisporus CU13 Laccase - Mediator System: A Statistical Study for Optimization via Response Surface Methodology

Aim: the present study aims to optimize Cibacron Blue 3G-A decolorization as a model dye through laccase ‎enzymatic biocatalysis presenting the role of HBT as a redox mediator via ‎RSM approach.‎ Study Design: RSM using Central Composite Design (CCD) was used in order to determine the most effective variables levels in Cibacron Blue 3G-A decolorization and to investigate their interactions. Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Research Division, National Research Centre (NRC), Cairo, Egypt, between August 2017 and January 2018. Methodology: The evaluation of Cibacron Blue 3G-A decolorization by A. bisporus CU13 crude laccase was conducted through different trials using a 1.5 mL reaction mixture containing different concentration of crude laccase, Cibacron Blue 3G-A, and HBT in 0.1 M sodium citrate buffer (pH 4.5) at room temperature for different incubation periods. Results: Hydroxybenzotriazole (HBT) as a mediator enhanced Cibacron Blue 3G-A decolorization levels significantly, where decolorization percentage caused by laccase enzyme alone were ‎11.92 and ‎23.78%, ‎whereas that caused by laccase HBT mediator system under the same conditions were 43.43 and ‎76.34% after 1 and 22 h of incubation, respectively. HBT concentration, dye concentration, enzyme activity, and incubation time were chosen as study variables to optimize Cibacron Blue 3G-A dye decolorization through RSM approach via central composite design (CCD). The optimum conditions for Cibacron Blue 3G-A decolorization were found to be under using 0.50 U/mL of Agaricus bisporus CU13 laccase, 92.19 ppm of Cibacron Blue 3G-A, and 1 mM of HBT in order to get decolorization percentage of 29.29% in 35 min. Conclusion: Agaricus bisporus CU13 crude laccase was used as a biocatalyst to decolorize Cibacron Blue 3G-A in presence of HBT as a mediator through utilizing the response surface methodology approach. HBT concentration, dye concentration, enzyme activity, ‎and incubation time affects the decolorization levels considerably.

High-throughput transcriptomic sequencing of Rheum palmatum L. seedlings and elucidation of genes in anthraquinone biosynthesis / 药学学报

Anthraquinones are not only the main active constituents but also the index components for the quality control of Rhei Radix et Rhizoma. To study the anthraquinone biosynthesis, Rheum palmatum L. seedlings were subjected to a high-throughput transcriptomic sequencing analysis by Illumina HiSeqTM 2000 150PE. The Illumina sequencing generated a total of 11.04 G clean data resulting in 736 309 74 clean reads, deposited in the sequence read archive (SRA accession SRP160030). Trinity do novo assembly yielded 93 646 unigenes, with an average of 1 108 nt. Functional annotation revealed that all unigenes were successfully annotated in the NR, NT, Swiss-port, PFAM, and KOG databases. GO enrichments showed that 57 subgroups were involved in biological process, cellular component, and molecular function. KEGG analysis indicated that 1 107 unigenes were implicated in 19 standard secondary metabolic pathways. 172 unigenes were analyzed to encode 28 key enzymes during the MVA, MEP, shikimic acid, and polyketide pathways related to anthraquinone biosynthesis. 125 CYP450 and 73 UGTs unigenes were related the modification of secondary metabolites in R. palmatum L. Furthermore, seven unigenes with full length cDNAs were successfully verified by RT-PCR and sequencing analyses. Then, MISA prediction produced a number of 18 885 simple sequence repeats (SSRs). Herein, the transcriptomic gene expression profiles of R. palmatum L. and candidate genes during the anthraquinone biosynthesis pathway were obtained for the first time. The results provided basic information for subsequent gene function characterization, secondary metabolic pathway analysis, and anthraquinone biosynthesis and regulation elucidation in R. palmatum L.

Protective effect and mechanism of total anthraquinone extract of Rhei Radix et Rhizoma on cerebral ischemia-reperfusion injury / 中成药

AIM To investigate the effect and mechanism of total anthraquinone extract of Rhei Radix et Rhizoma on cerebral ischemia-reperfusion injury and to provide relevant data references for its promising use in the management of cerebral ischemia-reperfusion injury.METHODS SD rats randomly assigned to model group,sham operation group,nimodipine group,total anthraquinone extract groups (high,medium and low dose),8 in each group,were orally administered with corresponding drugs daily for a week,with rats of the model group and sham operation group given the same volume of normal saline before the models established by middle cerebral artery occlusion (MCAO).MCAO started thirty minutes after final oral administration,and the induced ischemia went on for 1.5 h for a further reperfusion,24 h after which the neurological function score,brain index,brain water content and cerebral infarct volume were measured.Elisa kits were used to detect superoxide dismutase (SOD),Malondialdehyde (MDA),Nitric oxide (NO),interleukin-1 β (IL-1β),tumor necrosis factor (TNF),interleukin-6 (IL-6).RESULTS The total anthraquinone extract of Rhei Radix et Rhizoma significantly improved the neurological function score,decreased the brain index,brain water content,reduced the cerebral infarct volume (P ＜ 0.05),increased the activity of SOD in brain tissue (P ＜ 0.01),and reduced the levels of MDA and NO in brain tissue (P ＜0.01),and the levels of IL-6,TNF and IL-1β in serum (P ＜0.01) as well.CONCLUSION The obviously protective effect on rats' cerebral ischemia-reperfusion injury by total anthraquinone extract of Rhei Radix et Rhizoma may contribute to its inhibition of inflammatory response,and its existence as an antioxidant as well.

Simultaneous determination of 10 anthraquinone derivatives in Rhei Radix et Rhizoma based on UPLC / 中草药

Objective To establish a simultaneous quantitative analytical method of 10 anthraquinone derivatives in Rheum officinale based on ultra-high performance liquid chromatography (UPLC) for the quality evaluation and control. Methods The UPLC method was performed on a BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with a binary gradient mobile phase consisting of a mixture of methanol and 0.1% aqueous phosphoric acid with column temperature at 34 ℃. Using a gradient elution method; The flow rate was set at 0.20 mL/min; The injection volume was set at 2.0 μL; The detection wavelength was set at 410 nm. This study focused on the influence of analytical parameters on the separation efficiency of five anthraquinone glucosides in R. officinale. To obtain robust chromatographic resolution of the five anthraquinone glucosides, relatively important chromatographic parameters including the retention time, resolution, theoretical plate number, and symmetry factor were investigated by altering the column temperature and flow rate. Results The optimal column temperature was found to be 34 ℃, and the optimal flow rate was 0.20 mL/min. A new UPLC analytical method was developed for simultaneous quantification of 10 anthraquinone derivatives in R. officinale based on these optimal parameters. This UPLC-based analytical method was successfully applied to quantitative determination of 10 anthraquinone derivatives in Rheum tanguticum and Rheum palmatum. The analysis showed that there were significant content differences between aloe-emodin-8-O-β-D-glucoside (more than 60-fold) and rhein-8-O-β-D-glucoside (more than 77-fold) in the two species of rhubarbs tested. Conclusion The UPLC method established for the simultaneous determination of 10 anthraquinone derivatives in R. officinale based on the optimal parameters is accurate and sensitive, which will provide a more comprehensive evaluation for the chemical characterization of medicinal materials of R. officinale from different sources.

Preliminary study on antiplatelet aggregation of 10 anthraquinones in Rhei Radix et Rhizoma based on bioassay / 中草药

Objective To evaluate the ability of the antiplatelet aggregation of ten anthraquinone derivatives in Rhei Radix et Rhizoma by using bioassay method, and to screen for the indicators that can be used to control the quality of rhubarb wine processing product. Methods Platelet aggregation instrument was used to determine the platelet aggregation rate induced by ADP in vitro and calculate the antiplatelet aggregation rates of 10 anthraquinone derivatives (aloe-emodin, rhein, emodin, chrysophanol, physcion, aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside, chrysophanol-8-O-β-D-glucoside, and physcion-8-O-β-D-glucoside) at different concentrations. The biopotency was calculated by bioavailability software. In order to verify the accuracy of the bioassay results, the estimated inhibition constant of rhein and chrysophanol-8-O-β-D-glucoside in P2Y12 protein receptor were determined by molecular docking software. Results The bioassay results showed that the antiplatelet potencies of rhein and emodin were significantly higher than those of aloe-emodin, chrysophanol and physcion. Compared with the antiplatelet biopotency of aspirin, the antiplatelet potencies of rhein and emodin were 5.02 and 5.15 times higher than that of aspirin, which indicated that rhein and emodin have strong inhibitory effect on ADP-induced platelet aggregation. However, the antiplatelet potencies of aloe-emodin, chrysophanol and physcion were equivalent of that of aspirin. The antiplatelet potencies of aloe-emodin-8-O-β-D-glucoside, rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside, chrysophanol-8-O-β-D-glucoside, and physcion-8-O-β-D-glucoside were higher 4.13, 4.46, 9.31, 5.46, and 7.80 times than that of aspirin, respectively, which indicated that the five anthraquinone glucosides had a strong ability to antagonize ADP-induced platelet aggregation. The results of molecular docking showed that P2Y12 protein had different selectivity to 10 anthraquinone derivatives, especially for rhein, chrysophanol-8-O-β-D-glucoside, and the estimated Ki value were 5.73 and 2.51 μmol/L, respectively, indicating that rhein, chrysophanol-8-O-β-D-glucoside produced a strong inhibitory effect on P2Y12 protein at a lower concentration level. Moreover, the activity of chrysophanol-8-O-β-D-glucoside was stronger than rhein, which was consistent with their measured intensity of antiplatelet aggregation. Conclusion All results showed that there were some differences among antiplatelet potencies of 10 anthraquinone derivatives, and finally the screening of rhein, emodin can be used as evaluation indicators to control the quality of rhubarb wine processing product.

Chemical constituents from root of Lasianthus acuminatissimus (II) / 中草药

Objective To study the chemical constituents from the root of Lasianthus acuminatissimus. Methods The compounds were isolated and purified by silica gel column chromatography, Sephadex LH-20, ODS column chromatography, preparative HPLC and so on. Their structures were determined on the basis of physicochemical properties and their spectroscopic data, as well as literature. Results A total of 16 compounds were separated and identified as 4-methoxyphenol (1), 3-acetylbetulinic acid (2), palmitic acid (3), 3,8-dihydroxy-2-ethoxymethyl-1-methoxy-9,10-anthraquinone (4), 1,3-dihydroxy-2-ethoxymethyl-9,10-anthraquinone (5), 1,3-dihydroxy-2-methyl-9,10-anthraquinone (6), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (7), 3-hydroxy-1-methoxy-9,10- anthraquinone (8), 1,3-dihydroxy-2-methoxymethyl-9,10-anthraquinone (9), 4-hydroxymethyl-2-furaldehyde (10), butyl benzyl phthalate (11), lasianthurin A (12), isoscopletin (13), damnacanthol (14), uncargenin A (15), and lasianthuoside A (16). Conclusion Compounds 1-11 are isolated from the genus of Lasianthus for the first time.