Lipidomic investigation of the protective effects of Polygonum perfoliatum against chemical liver injury in mice / 中西医结合学报

OBJECTIVE@#Recent investigations have demonstrated that Polygonum perfoliatum L. can protect against chemical liver injury, but the mechanism behind its efficacy is still unclear. Therefore, we studied the pharmacological mechanism at work in P. perfoliatum protection against chemical liver injury.@*METHODS@#To evaluate the activity of P. perfoliatum against chemical liver injury, levels of alanine transaminase, lactic dehydrogenase, aspartate transaminase, superoxide dismutase, glutathione peroxidase and malondialdehyde were measured, alongside histological assessments of the liver, heart and kidney tissue. A nontargeted lipidomics strategy based on ultra-performance liquid chromatography quadrupole-orbitrap high-resolution mass spectrometry method was used to obtain the lipid profiles of mice with chemical liver injury and following treatment with P. perfoliatum; these profiles were used to understand the possible mechanisms behind P. perfoliatum's protective activity.@*RESULTS@#Lipidomic studies indicated that P. perfoliatum protected against chemical liver injury, and the results were consistent between histological and physiological analyses. By comparing the profiles of liver lipids in model and control mice, we found that the levels of 89 lipids were significantly changed. In animals receiving P. perfoliatum treatment, the levels of 8 lipids were significantly improved, relative to the model animals. The results showed that P. perfoliatum extract could effectively reverse the chemical liver injury and significantly improve the abnormal liver lipid metabolism of mice with chemical liver injury, especially glycerophospholipid metabolism.@*CONCLUSION@#Regulation of enzyme activity related to the glycerophospholipid metabolism pathway may be involved in the mechanism of P. perfoliatum's protection against liver injury. Please cite this article as: Peng L, Chen HG, Zhou X. Lipidomic investigation of the protective effects of Polygonum perfoliatum against chemical liver injury in mice. J Integr Med. 2023; 21(3): 289-301.

Estudio etnobotánico de Cataia (Pimenta pseudocaryophyllus (Gomes) Landrum) en el Parque Nacional de Superagui, Guaraqueçaba/PR/Brasil / Ethnobotany of Cataia (Pimenta pseudocaryophyllus (Gomes) Landrum) at National Park of Superagui, Guaraqueçaba/PR/Brazil

Cataia es una Myrtaceae de gran potencial aromático y medicinal. En la costa de Paraná hay registro de uso comercial creciente de sus hojas en la aromatización de aguardiente. Buscando comprender mejor la explotación de cataia desde una perspectiva histórica, así como elucidar el conocimiento ecológico de los moradores acerca del manejo, se presentan los resultados de una investigación etnobotánica realizada en Barra do Ararapira (PR/Brasil). La recolección de datos ha incluido entrevistas semiestructuradas y acompañamiento a campo de los extractivistas. Se ha revelado potencial de sostenibilidad, denotado por el apoyo institucional, en lo que se refiere a conferir legalidad a la actividad. La baja demanda actual se presenta como punto positivo, con la necesidad de monitoreo continuo de posibles daños, frente al posible aumento de la demanda. Sobre la cadena de valor, la inexistencia de intermediarios y la venta regular a comerciantes, caracterizan un componente de comercialización favorable.

Cataia is a Myrtaceae of great aromatic and medicinal potential. On the coast of Paraná there is a record of increasing commercial use of its leaves in the aromatization of brandy. In order to better understand the exploitation of cataia from a historical perspective, as well as elucidate the ecological knowledge of the inhabitants about the management, the results of an ethnobotanical investigation carried out in Barra do Ararapira are presented. Data collect has involved individual and group interviews and monitoring of all extractor's activities. It was elucidated a sustainability potential, denoted at first instance by institutional support to confer legality to the activity and for their implementation, and the community organizational capacity. As regards to the ecological question, the low demand now existent was considerate a positive aspect, but is highlight the importance of a continuous monitoring of possible damage. Regarding the value chain was evidenced the absence of intermediaries and the regular sale to traders of the region, featuring a favorable marketing component.

Polygonum hydropiper extract attenuates ethanol-induced gastric damage through antioxidant and anti-inflammatory pathways

The present study was conducted to investigate the underlying mechanisms and effective components of Polygonum hydropiper in ethanol-induced acute gastric mucosal lesions. The ethanol extract was purified on an AB-8 macroporous resin column and eluted with 60% ethanol and was then injected into the HPLC system for quantitative analysis. Sprague-Dawley rats were orally pretreated with P. hydropiper extract (PHLE; 50, 100, and 200 mg/kg) for 5 days and then absolute ethanol was administered to induce gastric mucosal damage. One hour after ethanol ingestion, the rats were euthanized and stomach samples were collected for biochemical analysis. Antioxidant enzymes and anti-inflammatory cytokines were quantified. Western blotting was used to detect the expression levels of proteins. Cell proliferation was assayed by CCK-8 assays. The proportion of total flavonoids in the final extract of P. hydropiper was 50.05%, which contained three major bioactive flavonoid constituents, including rutin, quercitrin, and quercetin. PHLE significantly increased cell viability and effectively protected human gastric epithelial cells-1 against alcohol-induced damage in vitro. PHLE pretreatment attenuated gastric mucosal injuries in a dose-dependent manner in rats, and increased the activity of superoxide dismutase, glutathione peroxidase, and glutathione, and decreased the levels of malondialdehyde in gastric tissue. Pretreatment with PHLE also reduced the generation of the pro-inflammatory cytokines tumor necrosis factor-α and interleukin-1β in gastric tissue by downregulating the expression of nuclear factor-kappa B. PHLE exerted protective effects against gastric injury through antioxidant and anti-inflammatory pathways. Flavonoids might be the main effective components of P. hydropiper against gastric mucosal injury.

Pharmacokinetic study of Polygonum orientale extract in H9c2 cells by UPLC-MS/MS / 中国中药杂志

A detection method of ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was established to detect concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in H9 c2 cells and applied to the pharmacokinetic study of Polygonum orientale extract in the cells. H9 c2 cells were treated with 100 μg·mL~(-1) P. orientale extract and then they and the corresponding nuclei, mitochondria and Golgi bodies were collected at the set time. After protein precipitation, UPLC-MS/MS was used to determine concentrations of isoorientin, orientin, quercetin, vitexin and kaempferol-3-O-β-D-glucoside in the whole cells and subcellular structures. Also, related pharmacokinetic parameters were calculated. The results showed that the peak time was 8 h for all these components. Orientin, vitexin, quercetin and isoorientin have high affinities to nuclei and mitochondria, while the affinity of kaempferol-3-O-β-D-glucoside is higher with mitochondria compared to nuclei. It is suggested that these chemical components of P. orientale may mainly act on nuclei or mitochondria to exert pharmacological effects of protecting cardiomyocytes.

Two new sucrose cinnamates from Polygonum lapathifolium var. salicifolium / 中国中药杂志

Two new sucrose cinnamates(1 and 2) along with nine known compounds(3-11) were isolated from ethanol extract of Polygonum lapathifolium var. salicifolium by silica gel column chromatography, ODS column chromatography and semi-preparative HPLC. Their structures were elucidated by extensive spectroscopic methods including 1 D-and 2 D-NMR experiments, as well as HR-ESI-MS analysis. Eleven compounds(7 sucrose cinnamates, 3 phenylpropanoids and 1 lactone) were obtained and their structures were identified as(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside(1),(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-(6-O-acetyl)-α-D-glucopyranoside(2),(3-O-feruloyl)-β-D-fructofuranosyl-(2→1)-(6-O-p-coumaroyl)-α-D-glucopyranoside(3), hydropiperoside(4), vanicoside C(5),(1,3-O-di-p-coumaroyl)-β-D-fructofuranosyl-(2→1)-(6-O-feruloyl)-α-D-glucopyranoside(6), vanicoside B(7),trans-p-hydroxycinnamic acid methyl ester(8), trans-p-hydroxycinnamic acid ethyl ester(9), methyl ferulate(10) and dimethoxydimethylphthalide(11), respectively. Compounds 1 and 2 were two new sucrose cinnamates, and compounds 1-11 were isolated from this plant for the first time. The antioxidant activities of the isolated compounds 1-9 were investigated by an oxygen radical absorbance capacity(ORAC) assay, and all nine compounds were found to show strong antioxidant activities. Among them, compound 6(10 μmol·L~(-1)) was the supreme one in antioxidant activities, with its ORAC value equivalent to(1.60±0.05) times of 50 μmol·L~(-1) Trolox.

Intestinal absorption characteristics of Polygonum orientale extract in normal and isoproterenol-induced myocardial ischemia model rats via everted intestinal sac models / 中国中药杂志

The present study is to investigate the absorption characteristics of the main components in Polygonum orientale extract in normal and isoproterenol-induced myocardial ischemia model rats with everted intestinal sac models. Intestinal sac fluid samples were collected in different part of intestine(duodenum, jejunum, ileum, colon) at different time after administration of different concentration of P. orientale extract(5.0,10.0, 20.0 mg·mL~(-1)). An UPLC-TQD method was employed for the determination of six components including orientin, isoorientin, vitexin, protocatechuic acid, kaempferol-3-O-β-D-glucoside and quercitrin in the intestinal sac samples. The absorption rate and cumulative absorption were calculated to analyze the intestinal absorption characteristics of six components in normal and myocardial ischemia model rats. The P-glycoprotein(P-gp) inhibitor was applied to investigate influence of intestinal absorption of six components in P. orientale extract. The results showed that the main absorption sites were concentrated on the duodenum at low concentration, while they were the colon at the medium concentration and the ileum at high concentration in control groups. In the condition of myocardial ischemia model, the main absorption sites focus on the ileum and jejunum at low concentration; the main absorption sites were in the ileum at the medium concentration and main absorption sites were the duodenum and ileum at high concentration. Compared with the normal group, the absorption rate and cumulative absorption of the six components significantly decreased in the model group. P-gp inhibitor markedly increased the absorption rate and cumulative absorption of six components in the model group, inferring that the 6 components may be the substrates of P-gp, and the mechanism needs further study. In this study, it is revealed that the six components of P. orientale extract can be absorbed into the intestinal sac, and it is an effective method to assess the intestinal absorption characteristics of P. orientale extract through everted intestinal sac model, providing data support for the clinical application and further development of P. orientale.

Similar change laws of content of nine components in four processing methods of Polygoni Multiflori Radix Praeparata / 中国中药杂志

To standardize the processing technology of Polygoni Multiflori Radix Praeparata and stabilize its quality, the similar change laws of Polygoni Multiflori Radix Praeparata with different processing methods and time were analyzed. The processing time of Polygoni Multiflori Radix Praeparata was studied at 24, 32, 40, 48 h, and 4 different processing methods were studied, namely stewed with black bean sauce, steamed, steamed with black bean sauce, and steamed with black bean sauce and rice wine. The content of stilbene glycosides and anthraquinones were determined by HPLC-DAD method. UV method was used to determine the content of polysaccharides, and HPLC-ELSD method was used to determine the monosaccharides and oligosaccharides. The comparative chart of content determination, cluster analysis and entropy weight TOPSIS model was used to find the similar change laws and time interval of different processing methods of Polygoni Multiflori Radix Praeparata. The results demonstrated that around 32 h, the content of nine components in Polygoni Multiflori Radix Praeparata with different processing methods had similar change laws, and the decoction pieces had a high quality, indicating that the four processing methods of Polygoni Multiflori Radix Praeparata are likely to be used as one type of decoction piece with the same name.

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.

Research progress of liver injury induced by Polygoni Mulitiflori Radix / 中国中药杂志

Polygoni Mulitiflori Radix, or dried root tuber of Polygonum multiflorum(PM), is a traditional Chinese tonic medicine, with the effect in nourishing liver and kidney, and benefiting blood essence and hair. It is widely used in clinical and healthcare products. In recent years, more and more reports about adverse reactions of root tuber of P.multiflorum and its preparations have been reported. Fortunately, there is also substantial progress in the experimental study on liver injury induced by PM. According to the literature review, the possible causes of liver injury were found to be the mixture of raw and processed PM and long-term high-dose administration. In addition, the liver injury induced by PM is idiosyncratic liver injury, and individual factors are also the important cause. At the same time, according to the literature reports, the effects of chemical components in different pathological animal models were summarized, finding that 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside was the main component for liver injury; based on the clinical manifestations of liver injury induced by PM, the effects of some chemical components on bilirubin and bile acid metabolism were analyzed. This paper reviews the study progress of liver injury induced by PM.

Isolation of dihydrobenzofuran derivatives from ethnomedicinal species Polygonum barbatum as anticancer compounds

BACKGROUND: Ethnomedicinally, the family Polygonaceae is famous for the management of cancer. Various species of this family have been reported with anticancer potentials. This study was designed to isolate anticancer compounds from ethnomedicinally important species Polygonum barbatum. METHODS: The column chromatography was used for the isolation of compounds from the solvent fraction of P. barbatum. The characterization of isolated compounds was performed by various spectroscopic techniques like UV, IR, mass spectrometry and 1D-2D NMR spectroscopy. Keeping in view the ethnomedicinal importance of the family, genus and species of P barbatum, the isolated compounds (1-3) were screened for anticancer potentials against oral cancer (CAL-27) and lungs cancer (NCI H460) cell lines using MTT assay. Active compound was further investigated for apoptosis by using morphological changes and flow cytometry analysis. In vivo anti-angiogenic study of the isolated compounds was also carried using chorioallantoic membrane assay. Docking studies were carried out to explore the mechanism of anticancer activity. RESULTS: Three dihydrobenzofuran derivatives (1-3) have been isolated from the ethyl acetate fraction of P. barbatum. The structures of isolated compounds were elucidated as methyl (2S,3S)-2-(3,4-dimethoxyphenyl)-4-((E)-3-ethoxy-3-oxoprop-1-en-1-yl)-7-methoxy-2,3-dihydrobenzo-furan-3-carboxylate (1), (E)-3-((2S,3S)-2-(3,4-dimethoxyphenyl)-7-methoxy-3-(methoxy carbonyl)-2,3-dihydrobenzofuran-4-yl)acrylic acid (2) and (2S,3 S)-4-((E)-2-carboxyvinyl)-2-(3,4-dimethoxyphenyl)-7-hydroxy-2,3-dihydrobenzofuran-3-carboxylic acid (3). The compound 1 was found to be more potent with IC50 of 48.52 ± 0.95 and 53.24 ± 1.49 against oral cancer cells as compared to standard drug (IC50 = 97.76 ± 3.44 µM). Both compound also inhibited lung cancer cells but at higher concentrations. Morphological and flow cytometry analysis further confirms that compound 1 induces apoptosis after 24 to 48 h treatment. In antiangiogenesis assay, compounds 1, 2 and 3 exhibited IC50 values of 8.2 ± 1.1,13.4 ± 1.1 and 57.7 ± 0.3 µM respectively. The docking studies revealed that the compounds under study have the potential to target the DNA and thymidylate synthase (TS). CONCLUSION: Based on its overwhelming potency against the tested cell lines and in angiogenesis assay, compound 1 can be further evaluated mechanistically and can be developed as anticancer drug candidate.

UPLC-Q-TOF-MS/MS analysis on prototypes components and metabolites of effective fractions of Polygonum orientale flower in rat serum and urine / 中国中药杂志

Ultra performance liquid chromatography coupled with time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) method was applied to analyze the prototypes and metabolites of the effective components of Polygonum orientale in SD rat serum and urine. The separation was performed on Agilent Eclipse Plus C_(18) column( 2. 1 mm×100 mm,1. 8 μm),with 0. 1% formic acid solution( A)-acetonitrile( B) as the mobile phase for gradient elution. Mass spectrometry data of biological samples were obtained under positive and negative electrospray ion mode. By comparing chromatogram differences between blank samples and drug treatment samples,prototype components and metabolites of the effective components of P. orientale extract were identified. The results showed that 12 metabolites were detected in serum and 26 metabolites in urine( including cross-components) of rats. The main metabolic pathways included hydrogenation,hydroxylation,glucuronidation,sulfation reaction,and methylation-glucuronidation,etc. The method established in this study was reliable and effective for studying the metabolic characteristics of the effective components of P. orientale in rats,and it can provide a reference for further studies on therapeutic material basis of this herb.

Effectual components of Polygonum ciliinerve protects against Staphylococcus aureus infection with immunomodulatory functions in C57BL/6 mice

Abstract Purpose: To investigate the efficacy and mechanisms of root tuber of Polygonum ciliinerve (Nakai) ohwi (rPC) which has been used to treat bacterial infection in traditional Chinese medicine. Methods: With the mouse model of Staphylococcus aureus (S. aureus) pneumonia, the phenotype of rPC treated mice, including body weight, mortality, lung slices and bacterial burden were evaluated. Furthermore, inflammatory factors in bronchoalveolar lavage (BAL) were determined by ELISA and the distribution of T cells in lung was assessed by immunofluorescence assay. Results: rPC treatment could dose-dependently reduce weight loss and mortality in S. aureus-infected mice. Upon 10 mg/ml rPC treatment, S. aureus-infected mice showed about 8 grams increase in body weight (P<0.001) and 50% enhancement in mortality. The integrity of lung tissue and bacterial burden were also improved by rPC treatment. Moreover, rPC was found to modulate the immune response in infection. Conclusion: rPC has therapeutic potential for S. aureus infections and pneumonia with immunomodulatory functions.

Chemical profiles and metabolite study of raw and processed Polygoni Multiflori Radix in rats by UPLC-LTQ-Orbitrap MS spectrometry / 中国天然药物

The raw and processed roots of Plygonum multiflorum Thunb (PM) are used to treat different diseases in clinical practice. In order to clarify the influence of processing, a comparative study of chemical substance analysis was carried out. As the xenobiotics with a high enough exposure in target organs being considered as the potential effective or toxicity components, an in vivo study was also implemented to characterize the constitutes and metabolites, and meanwhile, the factor of compatibility with black bean were also considered. As a result, a total of 148 compounds were detected in PM extracts and more than 40 compounds were only detected in the processed products, which were probably new components produced during the steaming process. In in vivo study, 7 prototype components and 66 metabolites were detected or tentatively identified, 24 of which were reported for the first time. Our results indicated that processing greatly changed the chemical composition of PM and influenced the disposition of the compounds in vivo. To the best of our knowledge, this was the first global comparative study of raw and processed PM. These results expanded our knowledge about the influence of processing of PM and provided the essential data for further efficacy or toxicity studies.

Effect of six components in Polygoni Multiflori Radix on regulation of CYP3A4 mediated by human pregnane X receptor / 中国中药杂志

This paper aimed to study the six chemical components of Polygoni Multiflori Radix (gallic acid, quercetin, luteolin, kaempferol, resveratrol, apigenin). By the established pregnane X receptor (human pregnant X receptor, PXR) CYP3A4 mediated drug induced rapid screening technique, the effect of chemical components on the cell activity was detected by MTS cell method, and the value of IC₅₀ was calculated. The dual luciferase reporter system was used to co-transfect PXR reporter gene expression vector containing transcriptional regulation and CYP3A4 with HepG2 cells, with 10 μmol·L⁻¹ rifampicin (RIF) as a positive control, and 10 μmol·L⁻¹ of ketoconazole (TKZ) as negative control. Gallic acid, quercetin, luteolin, kaempferol, apigenin, resveratrol(5, 10, 20 μmol·L⁻¹) were used to incubate for 24 h, and the luciferase activity was detected. The results showed that when plasmid pcDNA3.1 was co-transfected with pGL4.17-CYP3A4, gallic acid and resveratrol had an inhibitory effect on the regulation of CYP3A4, and quercetin, luteolin, kaempferol had an inductive effect on CYP3A4; when pcDNA3.14-PXR was co-transfected with pGL4.17-CYP3A4, quercetin, luteolin, kaempferol, apigenin, resveratrol had an inductive effect. To sum up, the 6 reported liver injury components had inhibitory or activating effects on CYP3A4. After PXR plasmid was involved, 5 components had an inductive effect on CYP3A4, and the inductive effects of 2 components were significantly different. In this experiment, we found that 2 kinds of potential liver injury components in Polygoni Multiflori Radix had been induced by CYP3A4, which was achieved through PXR regulation. It suggested that attention shall be paid to potential drug interactions when combined with Polygoni Multiflori Radix, so as to improve the safety and efficacy.

Chemical profiles and metabolite study of raw and processed Polygoni Multiflori Radix in rats by UPLC-LTQ-Orbitrap MS spectrometry / 中国天然药物

The raw and processed roots of Plygonum multiflorum Thunb (PM) are used to treat different diseases in clinical practice. In order to clarify the influence of processing, a comparative study of chemical substance analysis was carried out. As the xenobiotics with a high enough exposure in target organs being considered as the potential effective or toxicity components, an in vivo study was also implemented to characterize the constitutes and metabolites, and meanwhile, the factor of compatibility with black bean were also considered. As a result, a total of 148 compounds were detected in PM extracts and more than 40 compounds were only detected in the processed products, which were probably new components produced during the steaming process. In in vivo study, 7 prototype components and 66 metabolites were detected or tentatively identified, 24 of which were reported for the first time. Our results indicated that processing greatly changed the chemical composition of PM and influenced the disposition of the compounds in vivo. To the best of our knowledge, this was the first global comparative study of raw and processed PM. These results expanded our knowledge about the influence of processing of PM and provided the essential data for further efficacy or toxicity studies.

Endophytic fungi from tropical ethnoveterinary plants and their antibacterial efficacy against Pasteurella multocida Capsular Type A strain / Hongos endófitos de plantas tropicales etnoveterinarias y su eficacia antibacteriana contra cepas capsulares de Pasteurella multocida tipo A

AbstractPasteurella multocida is an important veterinary pathogen causing infections in animals and birds. Nowadays, different reports have described the severity of infections, increasing resistance of micro-organisms to antibiotics, and the contribution of ethnoveterinary practices towards the treatment of various ailments of animals. The aim of the present study was to investigate the antibacterial efficacy of the ethanolic extracts of endophytic fungi against P. multocida Capsular Type A strains. A total of six endophytic fungi were isolated from two tropical ethnoveterinary plants: Garcinia xanthochymus H. and Polygonum chinense L. The ethanolic extracts of the endophytic fungi were subjected to in vitro antimicrobial activity by the well diffusion method. Besides, we evaluated the treatment of mice with the potent fungal extract and observed the effects in different organs under electron microscopy. Our results showed that four fungi had antimicrobial activity against the selected pathogen. The best antibacterial activity was showed by the extract of the endophytic fungi, Glomerella magna isolated from G. xanthochymus, with a minimum inhibitory concentration of 46.9 µg/mL and minimum bactericidal concentration of 750 µg/mL. Treatment of mice with the potent fungal extract caused a considerable inhibitory effect on the pathogen growth in vital organs, results that was also confirmed by histopathological studies made by scanning electron microscopy. The present findings indicated that the endophytic fungi G. magna has the potential to provide an effective treatment against infections caused by Pasteurella multocida. However, the isolation of bioactive components needs further investigation. Rev. Biol. Trop. 64 (2): 733-745. Epub 2016 June 01.

ResumenPasteurella multocida es un importante patógeno veterinario que causa infecciones en animales y aves. Hoy en día, diferentes informes han descrito la gravedad de las infecciones, aumentando la resistencia de los microorganismos a los antibióticos, y la contribución de las prácticas etnoveterinarias hacia el tratamiento de diversas enfermedades de los animales. El objetivo del presente estudio fue investigar la eficacia antibacteriana de los extractos etanólicos de hongos endófitos contra cepas de P. multocida tipo capsular A. Un total de seis hongos endófitos fueron aisladas de dos plantas etnoveterinarias tropicales: Garcinia xanthochymus H. y Polygonum chinense L. Los extractos etanólicos de los hongos endófitos fueron sometidos a la actividad antimicrobiana in vitro por el método de difusión. Además, se evaluó el tratamiento de ratones con el extracto de hongos potente y observamos los efectos en diferentes órganos bajo el microscopio electrónico. Nuestros resultados mostraron que cuatro hongos tenían actividad antimicrobiana contra el patógeno seleccionado. La mejor actividad antibacteriana la mostró el extracto de los hongos endófitos, Glomerella magna aislado de G. xanthochymus, con una concentración inhibitoria mínima de 46.9 mg/ml y la concentración bactericida mínima de 750 mg/ml. El tratamiento de ratones con el extracto de hongos potente causó un considerable efecto inhibidor sobre el crecimiento de patógenos en órganos vitales, resultados que también fueron confirmados por estudios histopatológicos realizados por microscopía electrónica de barrido. Los presentes hallazgos indican que el hongos endófitos G. magna tienen el potencial de proporcionar un tratamiento eficaz contra las infecciones causadas por Pasteurella multocida. Sin embargo, el aislamiento de componentes bioactivos necesita más investigación.

Identification of Antioxidative Constituents from Polygonum aviculare using LC-MS Coupled with DPPH Assay

A method for simultaneously identifying antioxidative compounds was developed using time-based LC-MS coupled with DPPH assay regardless of the time consuming process. The methanolic extract of Polygonum aviculare (Polygonaceae) showed significant DPPH radical scavenging activity. Time-based DPPH assay for simultaneous identification of active compounds from the extracts of P. aviculare was used. Major peaks of ethyl acetate fraction of P. aviculare showed high DPPH radical scavenging activity. A simple phenolic compound (1) and six flavonoids (2-7) were isolated from the ethyl acetate fraction of P. aviculare by silica gel and sephadex LH-20 column chromatography. The structures of seven compounds were determined to be protocatechuic acid (1), catechin (2), myricitrin (3), epicatechin-3-O-gallate (4), avicularin (5), quercitrin (6), and juglanin (7) based on the analysis of the 1H-NMR, 13C-NMR and ESI-MS data. All compounds exhibited significant antioxidant activity on DPPH assay and active compounds were well correlated with predicted one.

Anti-Helicobacter pylori Compounds from Polygonum cuspidatum

Anti-Helicobacter pylori activity guided fractionation led to the isolation of five anthraquinones, two stilbenes and one naphthoquinone from the EtOAc fraction of Polygonum cuspidatum, using silica gel column chromatography, Sephadex-LH20, MPLC and recrystallization. The chemical structures were identified to be physcion (1), emodin (2), anthraglycoside B (3), trans-resveratrol (4), anthraglycoside A (5), polydatin (6), 2-methoxy-6-acetyl-7-methyljuglone (7) and citreorosein (8) by UV, ¹H-NMR, ¹³C-NMR and mass spectrometry. Anti-Helicobacter pylori activity including MIC values of each compound was evaluated. All of the isolates exhibited anti-H. pylori activity of which MIC values were lower than that of a positive control, quercetin. Compounds 2 and 7 showed potent growth inhibitory activity. Especially, a naphthoquinone, compound 7 displayed most potent antibacterial activity with MIC₅₀ value of 0.30 µM and MIC₉₀ value of 0.39 µM. Although anti-H. pylori activity of this plant was previously reported, this is the first report on that of compounds isolated from this species. From these findings, P. cuspidatum roots or its isolates may be useful for H. pylori infection and further study is needed to elucidate mechanism of action.

Ação genotóxica e antiproliferativa de Polygonum punctatum Elliott (Polygonaceae) sobre o ciclo celular de Allium cepa L / Genotoxic and antiproliferative action of Polygonum punctatum Elliott (Polygonaceae) on the cell cycle of Allium cepa L

RESUMO: A espécie Polygonum punctatum Elliott (Polygonaceae) é amplamente utilizada pela população como planta medicinal. O objetivo deste trabalho é o de avaliar o potencial genotóxico e mutagênico de P. punctatum utilizando raízes de bulbos e radículas em sementes germinadas de Allium cepa através do teste in vivo, e realizar comparações da extração do material vegetal por calor (infusões) e extração a frio (extrato). Para isso, foram preparadas dois tipos de soluções, infusões e extratos foliares de P. punctatum, em duas concentrações 0,4 g mL-1 e 2,4 g mL-1. A infusão foi preparada pela adição das folhas secas em água destilada fervente (100ºC), permanecendo por 10 minutos enquanto o extrato foi preparado através da maceração das folhas secas em água destilada fria. Para o teste em A. cepa foram utilizados, para cada tratamento, seis grupos de quatro bulbos e seis caixas gerbox com 50 sementes em cada caixa. Duas lâminas para cada tratamento foram obtidas através da técnica de esmagamento das raízes e coradas com orceína acética 2%. Foram contadas 2000 células por grupo de bulbos e 3000 células por grupo de sementes, observando-se a ocorrência de interrupções em metáfases, alterações cromossômicas estruturais, bem como a inibição ou aumento da divisão celular. Os valores do índice mitótico foram calculados e analisados estatisticamente pelo Teste χ2 (p≤0,05). Os resultados demonstraram que as infusões e os extratos de folhas apresentaram redução nos valores de índices mitóticos nas concentrações utilizadas em relação ao controle em água destilada. Foram identificadas alterações cromossômicas na divisão celular, tais como pontes anafásicas, em todas as concentrações de infusões e extratos indicando assim que P. punctatum possui atividade antiproliferativa e genotóxica.

ABSTRACT: The species Polygonum punctatum Elliott (Polygonaceae) is widely used by the Brazilian population as a medicinal plant. The aims of this study are to evaluate the genotoxic and mutagenic potential of P. punctatum, using its root bulbs and rootlets in germinated seeds of Allium cepa by in vivo testing, and to compare the extraction of plant material by heat (infusions) and cold (extract). Thus, two types of solutions - infusions and leaf extracts - of P. punctatum were prepared at the two concentrations of 0.4 g ml -1 and 2.4 g mL- 1. The infusion was prepared by addition of dry leaves in boiling distilled water (100ºC), remaining for 10 minutes, while the extract was prepared by maceration of dried leaves in cold distilled water. For the A. cepa, we used for each treatment six groups of six bulbs and six seedling boxes with 50 seeds each. Two slides for each treatment were obtained by the technique of crushing the roots, and they were stained with 2 % acetic orcein. For the analysis, 2000 cells per group of bulbs and 3000 cells per group of seeds were counted, and we noted the occurrence of interruptions in the metaphase, chromosomal aberrations, as well as inhibited or increased cell division. The values of the mitotic index were calculated and statistically analyzed by the χ2 test (p ≤ 0.05). The results showed that the infusions and extracts of leaves showed reduced values of mitotic indices in the concentrations used compared to the control in distilled water. Chromosomal alterations were identified in the cell division, in all concentrations of infusions and extracts, thus indicating that P. punctatum has an antiproliferative and genotoxic activity.

Intestinal absorption kinetics of Polygonum capitatum extract in rats / 中国中药杂志

A UPLC-ESI-MS/MS method was used to determinate the main active fractions gallic acid, protocatechuic acid, myricetrin, hyperoside and quercitrin in Polygonum capitatum extracts by in situ intestinal perfusion models; the absorption rate constants and cumulative penetration rate of absorption were calculated. The effect of different drug concentrations, different intestine segments, bile and P-gp inhibitors on the absorption mechanism of Gallic acid and other compositions in P. capitatum extracts. The experimental results showed that gallic acid, protocatechuic acid, myricetrin and quercitrin were observed saturated at high concentration (P < 0.05). Bile had significant inhibition effect on protocatechuic acid absorption and had promotion effect on myricetrin and hyperoside absorption (P < 0.05). P-gp inhibitor verapamil could significantly enhance the absorption of Protocatechuic acid (P < 0.05). The overall trend for absorption of various compositions was that small intestine > colon. This indicated that the absorption mechanism of P. capitatum extracts in rat intestine was in line with fist-order kinetics characteristics. The composition could be absorbed in all of the different intestinal segments, and the absorption was mainly concentrated in small intestine. The protocatechuic acid may be the substrate of P-gp.