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1.
Article in Chinese | WPRIM | ID: wpr-878960

ABSTRACT

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.


Subject(s)
Antioxidants , Cinnamates , Esters , Molecular Structure , Polygonum , Sucrose
2.
Article in Chinese | WPRIM | ID: wpr-878915

ABSTRACT

Caffeic acid and its oligomers are the main water-soluble active constituents of the traditional Chinese medicine(TCM) Arnebiae Radix. These compounds possess multiple biological activities such as antimicrobial, antioxidant, cardiovascular protective, liver protective, anti-liver fibrosis, antiviral and anticancer activities. The phenylpropanoid pathway in plants is responsible for the biosynthesis of caffeic acid and its oligomers. Glycosylation can change phenylpropanoid solubility, stability and toxic potential, as well as influencing compartmentalization and biological activity. In view of the important role played by de-glycosylation in the regulation of phenylpropanoid homeostasis, the biosynthesis of caffeic acid and its oligomers are supposed to be under the control of relative UDP-glycosyltransferases(UGTs). Through the data mining of Arnebia euchroma transcriptome, we cloned 15 full-length putative UGT genes. After recombinant expression using the prokaryotic system, the crude enzyme solution of the putative UGTs was examined for the glycosylation activities towards caffeic acid and rosmarinic acid in vitro. AeUGT_01, AeUGT_02, AeUGT_03, AeUGT_04 and AeUGT_10 were able to glycosylate caffeic acid and/or rosmarinic acid resulting in different mono-and/or di-glycosylated products in the UPLC-MS analyses. The characterized UGTs were distantly related to each other and divided into different clades of the phylogenetic tree. Based on the observation that each characterized UGT exhibited substrate or catalytic similarity with the members in their own clade, we supposed the glycosylation abilities towards caffeic acid and/or rosmarinic acid were evolved independently in different clades. The identification of caffeic acid and rosmarinic acid UGTs from A. euchroma could lead to deeper understanding of the caffeic acid oligomers biosynthesis and its regulation. Furthermore, these UGTs might be used for regiospecific glycosylation of caffeic acid and rosmarinic acid to produce bioactive compounds for potential therapeutic applications.


Subject(s)
Boraginaceae/genetics , Caffeic Acids , Chromatography, Liquid , Cinnamates , Cloning, Molecular , Depsides , Glycosyltransferases/genetics , Phylogeny , Tandem Mass Spectrometry
3.
Arq. bras. oftalmol ; 83(4): 305-311, July-Aug. 2020. graf
Article in English | LILACS | ID: biblio-1131600

ABSTRACT

ABSTRACT Purpose: Collagen deposition and myofibroblast differentiation are critical factors related to excessive scarring in ocular surgeries. This study evaluated the anti-fibrotic activity of rosmarinic acid on rabbit Tenon's capsule fibroblasts stimulated with transforming growth factor- β2. Methods: Primary cultures of rabbit Tenon's capsule fibroblasts were treated with various concentrations of rosmarinic acid for 12 h, in the presence and absence of transforming growth factor-β2. After 48 h, the proliferation index of rabbit Tenon's capsule fibroblasts and the differentiation of myofibroblasts were investigated through immunofluorescence staining for proliferating cell nuclear antigen and alpha smooth muscle actin. An automated cell counter and colorimetric metabolic activity assay were used to evaluate cell number and viability. Collagen expression and production were determined by quantitative real-time polymerase chain reaction and hydroxyproline assay, respectively. Results: Unstimulated rabbit Tenon's capsule fibroblasts treated with any concentration of rosmarinic acid exhibited diminished collagen expression (p<0.01) but showed no differences in proliferation index. Transforming growth factor-β2 exposure induced myofibroblast differentiation and increased collagen production. Exposure to rosmarinic acid at 1.0 and 3.0 µM concentrations reduced the proliferation index (p<0.02), as well as the collagen expression and hydroxyproline content (p<0.05). Exposure to 3.0 µM rosmarinic acid reduced viability (p=0.035) in unstimulated rabbit Tenon's capsule fibroblasts and cell numbers (p=0.001) in both stimulated and unstimulated rabbit Tenon's capsule fibroblast cultures. Conclusions: Exposure to 1.0 µM rosmarinic acid was noncytotoxic and led to reduced collagen expression and proliferation of stimulated rabbit Tenon's capsule fibroblasts. These findings suggest that rosmarinic acid is a relatively non-injurious anti-fibrotic compound to rabbit Tenon's capsule fibroblasts, with potential application as an adjunctive agent in ocular procedures, particularly in glaucoma surgeries.


RESUMO Objetivo: A deposição de colágeno e a diferenciação de miofibroblastos são fatores chaves relacionados à cicatrização excessiva em cirurgias oculares. Este estudo avaliou a atividade anti-fibrótica do ácido rosmarínico nos fibroblastos da cápsula de Tenon de coelhos estimulados com o fator de crescimento transformador-β2. Métodos: Culturas primárias de fibroblastos da cápsula de Tenon de coelhos foram tratadas com várias concentrações de ácido rosmarínico por 12h, na presença e na ausência do fator de crescimento transformador-β2. Após 48h, o índice de proliferação dos fibroblastos da cápsula de Tenon de coelhos e a diferenciação dos miofibroblastos foram investigados por coloração por imunofluorescência para proliferação de antígeno nuclear celular e α-actina de músculo liso, respectivamente. Um contador automático de células e um ensaio de atividade metabólica colorimétrica foram utilizados para avaliar o número e a viabilidade das células. A expressão e produção do colágeno foram determinadas por reação quantitativa em cadeia da polimerase em tempo real e ensaio de hidroxiprolina, respectivamente. Resultados: Fibroblastos da cápsula de Tenon de coelhos não estimulados tratados com qualquer concentração de ácido rosmarínico exibiram diminuição de colágeno (p<0,01), mas não mostraram diferenças no índice de proliferação. A exposição ao fator de crescimento transformador-β2 induziu a diferenciação de miofibroblastos e aumentou a produção de colágeno. A exposição ao ácido rosmarínico nas concentrações de 1,0 e 3,0 µM reduziu o índice de proliferação (p<0,02), bem como a expressão de colágeno e a quantificação de hidroxiprolina (p<0.05). A exposição a 3,0 µM de ácido rosmarínico reduziu a viabilidade (p=0,035) de fibroblastos da cápsula de Tenon de coelhos não estimulados e o número de células (p=0,001) em culturas de fibroblastos da cápsula de Tenon de coelhos estimuladas e não estimuladas. Conclusões: A exposição ao ácido rosmarínico 1,0 µM foi não citotóxica e levou à expressão reduzida de colágeno e menor proliferação de fibroblastos da cápsula de Tenon estimulados pelo fator de crescimento transformador-β2. Esses achados sugerem que o ácido rosmarínico é um composto antifibrótico relativamente não lesivo aos fibroblastos da cápsula de Tenon de coelhos, com potencial aplicação como agente adjuvante em procedimentos oculares, particularmente em cirurgias de glaucoma.


Subject(s)
Animals , Tenon Capsule , Rabbits , Cells, Cultured , Glaucoma , Cinnamates , Depsides , Fibroblasts
4.
Article in Chinese | WPRIM | ID: wpr-880798

ABSTRACT

OBJECTIVE@#To evaluate the effect of rosmarinic acid (RA) on mitophagy and hypertrophy of cardiomyocytes exposed to high glucose (HG).@*METHODS@#Rat cardiomyocytes (H9c2) exposed to HG (25 mmol/L) were treated with 50 μmol/L RA or with both RA treatment and Parkin siRNA transfection, with the cells cultured in normal glucose (5.5 mmol/L) and HG as the controls. The expressions of PINK1, Parkin and LC3II/LC3I in the cells were detected by Western blotting. The formation of mitochondrial autophagosomes was observed by transmission electron microscope. Flow cytometry was employed to detect the level of reactive oxygen species (ROS) and apoptotic rate of the cells. The activities of respiratory chain complex enzymes were measured by spectrophotometry. Fluorescence enzyme labeling and @*RESULTS@#RA treatment significantly increased the expression levels of PINK1, Parkin and LC3-II/I (@*CONCLUSIONS@#RA can protect rat cardiomyocytes against oxidative stress injury and cardiomyocyte hypertrophy induced by HG by activating Parkin-mediated mitophagy.


Subject(s)
Animals , Cinnamates , Depsides , Glucose , Hypertrophy , Mitophagy , Myocytes, Cardiac , Protein Kinases , Rats , Reactive Oxygen Species , Ubiquitin-Protein Ligases/genetics
5.
Acta cir. bras ; 34(11): e201901102, Nov. 2019. tab, graf
Article in English | LILACS | ID: biblio-1054682

ABSTRACT

Abstract Purpose: To investigate the effect of Picroside II on testicular ischemia and reperfusion (l/R) injury and the underlying mechanism. Methods: Sprague-Dawley rats were randomly divided into 4 groups: sham operated group (Sham), Sham with Picroside II treatment group (Sham+ Pic II), l/R group (l/R) and l/R with Picroside II treatment group (I/R+ Pic II). l/R model was established by rotating the left testis 720° in a clock-wise direction for 4 hours. The histopathologic and spermatogenetic evaluation was performed. The apoptosis changes and the levels of HO-1 (heme oxygenase-1), MPO (myeloperoxidase), NOX (NADPH oxidase), SOD (superoxide dismutase), XO (xanthine oxidase) and NOS (nitric oxide synthase) were measured. Results: The seminiferous tubules were damaged in l/R rats, but Picroside II alleviated the changes induced by l/R. The increased level of apoptosis was decreased by Picroside II (P=0.01, 9.05±0.35 vs. 4.85±0.25). The activities of HO-1, MPO, NOX, XO and MDA content were increased and the SOD activity was decreased in l/R (P<0.05) and could be reversed by Picroside II (P=0.03, 405.5±7.5 vs. 304±17U/mgprot; P=0.02, 0.99±0.05 vs. 0.52±0.04 mgprot; P=0.01, 260+7 vs. 189±2 mgprot; P=0.04, 10.95+0.55 vs. 8.75+0.35 U/mgprot; P=0.045, 6.8+0.7 vs. 3.75+0.35 mgprot; P=0.04, 44.5+3.5 vs. 57.5+3.5 mgprot). Western blot showed that the expression of iNOS, nNOS and eNOS were increased in l/R (P<0.05); however, they were decreased after Picroside II treatment (P<0.05). Conclusion: Picroside II attenuated testicular I/R injury in rats mainly through suppressing apoptosis and oxidative stress through reduction of nitric oxide synthesis.


Subject(s)
Animals , Male , Testis/blood supply , Reperfusion Injury/prevention & control , Cinnamates/pharmacology , Apoptosis/drug effects , Oxidative Stress/drug effects , Iridoid Glucosides/pharmacology , Nitric Oxide/biosynthesis , Reperfusion Injury/metabolism , Reperfusion Injury/pathology , Random Allocation , Blotting, Western , Rats, Sprague-Dawley , Peroxidase/analysis , In Situ Nick-End Labeling , Heme Oxygenase-1/analysis , Malondialdehyde/analysis , NADP/analysis
6.
Bol. latinoam. Caribe plantas med. aromát ; 18(1): 58-70, ene. 2019. tab, ilus
Article in English | LILACS | ID: biblio-1007470

ABSTRACT

The aim of this study was to evaluate the concentration and chemical composition of the essential oil the leaves of basil cultivars and hybrids cultivated in different cropping seasons: dry season and rainy season. The variables evaluated were the content and composition of essential oils in the two seasons. The essential oil content ranged from 0.66% to 3.21% in the dry season and from 0.80% to 4.20% in the rainy season. The major compounds found among the genotypes were linalool, methyl chavicol, neral, geranial, eugenol, and methyl (E)- cinnamate, defining the formation of five groups in each season, classified in the following chemotypes: methyl chavicol (Group 1), citral (neral+geranial) (Group 2), methyl cinnamate (Group 3), linalool (Group 4), and intermediate linalool (Group 5). All the traits evaluated had heritability (h ) greater than 95% and high CVg/CVe ratio values. The cropping season affected the content and chemical compositions of basil essential oil.


El objetivo de este estudio fue evaluar la concentración y composición química del aceite esencial las hojas de cultivares de albahaca e híbridos cultivados en diferentes temporadas de cultivo: estación seca y estación lluviosa. Las variables evaluadas fueron la contenido y la composición de los aceites esenciales en las dos estaciones. La contenido de aceite esencial varió de 0.66% a 3.21% en la estación seca y de 0.80% a 4.20% en la estación lluviosa. Los principales compuestos encontrados entre los genotipos fueron linalool, metilchavicol, neral, geranial, eugenol y metil (E)-cinamato, definiendo la formación de cinco grupos en cada estación, clasificados en los siguientes quimiotipos: metil chavicol (Grupo 1), citral (neral + geranial) (Grupo 2), cinamato de metilo (Grupo 3), linalool (Grupo 4) y linalol intermedio (Grupo 5). Todos los rasgos evaluados mostraron una heredabilidad (h ) mayor que el 95% y altos valores de relación CVg/CVe. La temporada de cultivo afectó la contenido y las composiciones químicas del aceite esencial de albahaca.


Subject(s)
Seasons , Oils, Volatile/chemistry , Plant Leaves/chemistry , Ocimum basilicum , Eugenol/analysis , Cinnamates/analysis , Monoterpenes/analysis , Anisoles/analysis
7.
Rev. Soc. Bras. Med. Trop ; 51(6): 849-853, Nov.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-1041494

ABSTRACT

Abstract INTRODUCTION Leishmaniasis, Chagas disease, and malaria cause morbidity globally. The drugs currently used for treatment have limitations. Activity of cinnamic acid analogs against Leishmania spp., Trypanosoma cruzi, and Plasmodium falciparum was evaluated in the interest of identifying new antiprotozoal compounds. METHODS In vitro effects of analogs against L. braziliensis, L. infantum chagasi, T. cruzi, and P. falciparum, and hemolytic and cytotoxic activities on NCTC 929 were determined. RESULTS Three analogs showed leishmanicidal and tripanocidal activity. No antiplasmodial, hemolytic, or cytotoxic activity was observed. CONCLUSIONS Antiprotozoal activity of analogs against L. infantum braziliensis, L. infantum chagasi, and T. cruzi was demonstrated.


Subject(s)
Plasmodium falciparum/drug effects , Trypanosoma cruzi/drug effects , Cinnamates/pharmacology , Leishmania/drug effects , Antiprotozoal Agents/pharmacology , Cinnamates/chemistry , Parasitic Sensitivity Tests , Antiprotozoal Agents/chemistry
8.
Article in Chinese | WPRIM | ID: wpr-775391

ABSTRACT

To establish the high performance liquid chromatography (HPLC) fingerprint for Digeda-4 decoction (DGD-4D), determine the contents of aesculetin, geniposide, picroside Ⅰ, picroside Ⅱ and ellagicacid in DGD-4D, and provide the scientific foundation for quality control of DGD-4D. The analysis was performed on Diamonsil(2) C₁₈ (4.6 mm×250 mm,5 μm) column, with methanol-0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. The flow rate was 1.0 mL·min⁻¹; injection size was 10 μL; temperature was maintained at 30 °C, and the detection wavelength was set at 254 nm. The common mode of DGD-4D HPLC fingerprint was established, and the hidden information was analyzed by Chemometrics. Chromatographic peaks for DGD-4D were identified by HPLC and quantitative analysis was conducted for characteristic peaks. There were 17 common peaks in the fingerprints and the similarity of the fingerprints was over 0.9 in all 15 batches. The samples were broadly divided into four kinds by principal component analysis and clustering analysis. Four marker compounds were verified by partial least squares discriminant analysis, and No. 9, 12 and 14 peaks were identified as geniposide, picroside Ⅱ, and picroside Ⅰ respectively. The average recoveries were in the range of 95.91%-97.31%. The HPLC fingerprint method for content determination is reliable, accurate, rapid, simple, and reproducible, and can be used as one of the effective methods to control the quality of DGD-4D.


Subject(s)
Chromatography, High Pressure Liquid , Cinnamates , Drugs, Chinese Herbal , Reference Standards , Iridoid Glucosides , Iridoids , Methanol , Principal Component Analysis , Quality Control
9.
Bol. latinoam. Caribe plantas med. aromát ; 16(4): 385-397, jul. 2017. ilus, tab
Article in English | LILACS | ID: biblio-882137

ABSTRACT

Myrcianthes is a Myrtaceous genus of flowering plants of about 30 to 40 species, distributed in the American continent. The aim of this work was to study the chemical composition of the foliar essential oil from M. fragrans growing wild in central Costa Rica. The essential oil was obtained through the steam distillation process in a Clevenger type apparatus. The chemical composition of the oil was performed by capillary gas chromatography with a flame detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS) using the retention indices on a DB-5 type capillary column in addition to mass spectral fragmentation patterns. A total of 98 compounds were identified, accounting for 98.8% of the total amount of the oil. The major constituents in the leaf oil were (E)-methyl cinnamate (39.6%), limonene (34.6%), α-pinene (6.8%), and linalool (6.8%). This is the first report of (E)-methyl cinnamate occurring in oils of this plant genus. These findings appear to suggest a new chemotype of M. fragrans.


Myrcianthes (Myrtaceae) consta de 30 a 40 especies, distribuidas en el continente americano. El objetivo del presente trabajo consistió en identificar la composición química del aceite esencial contenido en las hojas de M. fragrans, planta que crece en forma silvestre en el Valle Central de Costa Rica. La extracción del aceite se efectuó mediante el método de hidrodestilación usando un equipo de Clevenger modificado. La composición química del aceite se analizó mediante las técnicas de cromatografía gaseoso-líquida con detector de ionización de llama (GC-FID) y de cromatografía gaseoso-líquida acoplada a un detector de masas (GC-MS). Se utilizaron índices de retención obtenidos en una columna capilar tipo DB-5 y se compararon con los patrones de iones de fragmentación de masas. Se identificaron en total 98 compuestos, correspondientes a un 98.8% de los constituyentes totales. Los componentes mayoritarios del aceite resultaron ser (E)-cinamato de metilo (39.6%), limoneno (34.6%), α-pineno (6.8%) y linalol (6.8%). Este es el primer informe de la aparición de (E)-cinamato de metilo en aceite de hojas de este género de plantas. Los datos obtenidos parecen sugerir un nuevo quimiotipo de M. fragrans.


Subject(s)
Cinnamates/analysis , Myrtaceae/chemistry , Oils, Volatile/chemistry , Plant Leaves/chemistry , Terpenes/analysis , Chromatography, Gas/methods , Costa Rica , Cyclohexanes/analysis
10.
Article in English | WPRIM | ID: wpr-812040

ABSTRACT

Salvia miltiorrhiza is a medicinal plant widely used in the treatment of cardiovascular and cerebrovascular diseases. Hydrophilic phenolic acids, including rosmarinic acid (RA) and lithospermic acid B (LAB), are its primary medicinal ingredients. However, the biosynthetic pathway of RA and LAB in S. miltiorrhiza is still poorly understood. In the present study, we accomplished the isolation and characterization of a novel S. miltiorrhiza Hydroxyphenylpyruvate reductase (HPPR) gene, SmHPPR, which plays an important role in the biosynthesis of RA. SmHPPR contained a putative catalytic domain and a NAD(P)H-binding motif. The recombinant SmHPPR enzyme exhibited high HPPR activity, converting 4-hydroxyphenylpyruvic acid (pHPP) to 4-hydroxyphenyllactic acid (pHPL), and exhibited the highest affinity for substrate 4-hydroxyphenylpyruvate. SmHPPR expression could be induced by various treatments, including SA, GA, MeJA and Ag, and the changes in SmHPPR activity were correlated well with hydrophilic phenolic acid accumulation. SmHPPR was localized in cytoplasm, most likely close to the cytosolic NADPH-dependent hydroxypyruvate reductase active in photorespiration. In addition, the transgenic S. miltiorrhiza hairy roots overexpressing SmHPPR exhibited up to 10-fold increases in the products of hydrophilic phenolic acid pathway. In conclusion, our findings provide a new insight into the synthesis of active pharmaceutical compounds at molecular level.


Subject(s)
Amino Acid Sequence , Benzofurans , Biosynthetic Pathways , Genetics , Cinnamates , Depsides , Gene Expression Regulation, Plant , Genetics , Oxidoreductases , Genetics , Phenylpropionates , Metabolism , Phenylpyruvic Acids , Metabolism , Phylogeny , Plant Proteins , Genetics , Metabolism , Plant Roots , Chemistry , Genetics , Metabolism , Plants, Genetically Modified , Recombinant Proteins , Salvia miltiorrhiza , Chemistry , Genetics , Metabolism , Sequence Alignment
11.
Braz. j. med. biol. res ; 50(12): e6145, 2017. tab, graf
Article in English | LILACS | ID: biblio-888968

ABSTRACT

Chronic systemic inflammation and repetitive damage of vascular endothelia by incompatible dialysis system are probable causes of cardiovascular disease in patients on dialysis. The present study aimed to assess in vitro biocompatibility and anti-inflammatory effect of hemodialysis fluid supplemented with rosmarinic acid (RA) using human umbilical vein endothelial cells (HUVEC). HUVECs (5×106 cells/mL) were pre-exposed to 1 μg/mL of lipopolysaccharides (LPS) and incubated with RA-supplemented hemodialysis fluid (HDF). Cytotoxicity was assessed qualitatively by morphologic assessment and quantitatively by MTT assay. Expressions of proinflammatory mediators were assessed using quantitative real-time PCR and production of NO was quantified. Phosphorylation of AKT and nuclear localization of nuclear factor kappa B (NF-κB) were examined using western blotting. Exposure of HUVECs to RA-supplemented HDF had no influence on morphology and viability. Inhibition of proinflammatory mediator production in HUVECs by RA supplementation to HDF was significant in a dose-dependent manner. Exposure to RA-supplemented HDF resulted in a decrease in nitric oxide synthase expression and reduction of NO production in LPS-stimulated HUVECs. RA supplementation of HDF suppressed Akt activation in LPS-stimulated HUVECs. In addition, the level of cellular IκB was increased in parallel to a reduced nuclear translocation of NF-κB in LPS-induced endothelial cells. Our results suggest that RA-supplemented HDF is biocompatible and significantly suppressed inflammation induced in endothelial cells. In this respect, the use of HDF supplemented with RA could alleviate inflammation and improve long-term treatment of patients with renal failure on dialysis. Further clinical studies are required to confirm the effects.


Subject(s)
Humans , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Biocompatible Materials/pharmacology , Cinnamates/pharmacology , Depsides/pharmacology , Hemodialysis Solutions/pharmacology , Human Umbilical Vein Endothelial Cells/drug effects , Inflammation/drug therapy , Analysis of Variance , Cell Survival/drug effects , Cells, Cultured , Cytokines/analysis , Cytokines/drug effects , Formazans , Hemodialysis Solutions/chemistry , Human Umbilical Vein Endothelial Cells/metabolism , Immunoblotting , Inflammation/metabolism , Lipopolysaccharides , NF-kappa B/analysis , Nitric Oxide/analysis , Phosphorylation , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Tetrazolium Salts
12.
Journal of Breast Cancer ; : 358-371, 2016.
Article in English | WPRIM | ID: wpr-28543

ABSTRACT

PURPOSE: The protein kinase C (PKC) family of serine-threonine kinases plays an important role in cancer cell progression. Thus, molecules that target PKC have potential as anticancer agents. The current study aims to understand the treatment of breast cancer cells with alkyl cinnamates. We have also explored the mechanistic details of their anticancer action and the underlying molecular signaling. METHODS: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to measure the viability of MDAMB-231 breast cancer cells to assess the anticancer activity of these compounds. In addition, flow cytometry was performed to study the effect of alkyl cinnamates on the cell cycle and apoptosis. Immunoblotting and immunofluorescence techniques were performed to study PKC translocation, cytochrome c release, and modulation of the mitochondrial membrane potential in breast cancer cells targeted with alkyl cinnamates. RESULTS: The PKC agonist DM-2-8 translocated 16.6%±1.7% PKCα from cytosol to the plasma membrane and showed excellent anticancer activity with an half maximal inhibitory concentration (IC50) of 4.13±0.27 µg/mL against cancer cells. The treated cells had an abnormal morphology and exhibited cell cycle defects with G2/M arrest and reduced S phase. Cancer cells treated with DM-2-3, DM-2-4, or DM-2-8 underwent apoptosis as the major pathway of cell death, further confirmed by genomic DNA fragmentation. Furthermore, the mitochondrial membrane potential was perturbed, indicating involvement of the mitochondrial pathway of apoptosis. Immunolocalization studies revealed cytochrome c release from mitochondria to cytosol. Cancer cells treated with DM-2-8 and curcumin showed activation of caspase-9 and caspase-3 as downstream molecular components of the apoptotic pathway. Alkyl cinnamates also caused oxidative stress, which regulates the apoptotic machinery (DNA fragmentation), cell death, and morphological abnormalities in cancer cells. CONCLUSION: Alkyl cinnamates specifically target cancer cells through induction of PKC translocation and the mitochondrial pathway of apoptosis, and could be promising anticancer drugs.


Subject(s)
Antineoplastic Agents , Apoptosis , Breast Neoplasms , Breast , Caspase 3 , Caspase 9 , Caspases , Cell Cycle , Cell Death , Cell Membrane , Cinnamates , Curcumin , Cytochromes c , Cytosol , DNA Fragmentation , Flow Cytometry , Fluorescent Antibody Technique , Humans , Immunoblotting , Membrane Potential, Mitochondrial , Mitochondria , Oxidative Stress , Protein Kinase C , Protein Kinases , Protein-Serine-Threonine Kinases , S Phase
13.
Article in Chinese | WPRIM | ID: wpr-328284

ABSTRACT

<p><b>OBJECTIVE</b>To explore the neuroprotective effect and mechanism of picroside II on extracellular regulated protein kinases1/2 (ERK1/2) signal transduction pathway in cerebral ischemia injuryrats. METHODS The middle cerebral artery occlusion (MCAO) model was established by inserting a monofilament into middle cerebral artery. Totally 96 successfully modeled Wistar rats were divided into the modelgroup, the treatment (picroside II) group, the Lipopolysachcaride (LPS) group, and the U0126 group according to random digit table. Each group was further divided into 3 subgroups, i.e. 6, 12, and 24 h sub-groups. Picroside II (20 mg/kg) was peritoneally injected to rats in the treatment group 2 h after ischemia.LPS (20 mg/kg) and Picroside II (20 mg/kg) were peritoneally injected to rats in the LPS group 2 h after ischemia. U0126-EtOH (20 mg/kg)and Picroside II (20 mg/kg) were peritoneally injected to rats in the U0126group 2 h after ischemia. Equal volume of normal saline was peritoneally injected to rats in the control groupand the model group. The neurobehavioral function was evaluated by modified neurological severity score(mNSS) test. The structure of neurons was observed using hematoxylin-eosinstaining (HE) staining. Theapoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The expression of phosphorylated extracellular signal-regulated protein kinase1,2 (pERK1,2) in cortex was detected using immunohistochemistry (IHC) and Western blot.</p><p><b>RESULTS</b>After cerebral ischemia injury neurological impairment score increased, the damage of neuron in the cortical area was aggravated, apoptotic cells increased in the model group as time went by. The expression of pERK1/2 increased more significantly in the model group than in the control group (P <0.05). The damage of neuron in the cortical area was milder, while apoptotic cells decreased, the expression of pERK1f2 obviously decreased more in the treatment group and the U0126 group (P < 0.05). The early damage of neuron in the cortical area was more severe, apoptotic cells and the expression of pERK12 were comparatively higher in early stage of the LPS group, but the expression of pERK1/2 was somewhat decreased in late stage.</p><p><b>CONCLUSIONS</b>Activating ERK12 pathway could mediate apoptosis and inflammatory reactions of neurons after cerebral ischemia injury. Picroside II could protect the nerve system possibly through reducing activation of ERKI2 pathway, inhibiting apoptosis of neurons and inflammation reaction.</p>


Subject(s)
Animals , Apoptosis , Brain Ischemia , Drug Therapy , Cinnamates , Pharmacology , Infarction, Middle Cerebral Artery , Drug Therapy , Iridoid Glucosides , Pharmacology , MAP Kinase Signaling System , Neurons , Pathology , Neuroprotective Agents , Pharmacology , Random Allocation , Rats , Rats, Wistar
14.
Article in English | WPRIM | ID: wpr-310875

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of berberine (BBR) and cinnamic acid (CA), the main active components in Jiaotai Pill (, JTP), on palmitic acid (PA)-induced intracellular triglyceride (TG) accumulation in NIT-1 pancreatic β cells.</p><p><b>METHODS</b>Cells were incubated in culture medium containing PA (0.25 mmol/L) for 24 h. Then treatments with BBR (10 μmol/L), CA (100 μmol/L) and the combination of BBR and CA (BBR+CA) were performed respectively. Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by colorimetric assay. The expression of adenosine monophosphate-activated protein kinase (AMPK) protein and its downstream lipogenic and fatty acid oxidation genes, including fatty acid synthase (FAS), acetyl-coA carboxylase (ACC), phosphorylation acetyl-coA carboxylase (pACC), carnitine acyl transferase 1 (CPT-1) and sterol regulating element binding protein 1c (SREBP-1c) were determined by Western blot or real time polymerase chain reaction.</p><p><b>RESULTS</b>PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 cells. PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as pACC and CPT-1. Meanwhile, AMPK downstream lipogenic genes including SREBP-1c mRNA, FAS and ACC protein expressions were increased. Treatments with BBR and BBR+CA, superior to CA, significantly reversed the above genes changes in NIT-1 pancreatic β cells. However, the synergistic effect of BBR and CA on intracellular TG content was not observed in the present study.</p><p><b>CONCLUSION</b>It can be concluded that in vitro, BBR and BBR+CA could inhibit PA-induced lipid accumulation by decreasing lipogenesis and increasing lipid oxidation in NIT-1 pancreatic β cells.</p>


Subject(s)
AMP-Activated Protein Kinases , Metabolism , Animals , Berberine , Chemistry , Pharmacology , Cell Line , Cinnamates , Chemistry , Pharmacology , Fatty Acids , Metabolism , Gene Expression Regulation , Insulin-Secreting Cells , Metabolism , Intracellular Space , Metabolism , Lipogenesis , Genetics , Mice , Oxidation-Reduction , Palmitic Acid , Toxicity , Triglycerides , Metabolism
15.
Article in English | WPRIM | ID: wpr-812599

ABSTRACT

The present study was designed to analyze the major constituents in Prunellae Spica and establish a method for simultaneous determination of two constituents contained in Prunellae Spica. High performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-QTOF-MS/MS) technique was used to identify the constituents in the extractive of Prunellae Spica. High performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) was used to simultaneously quantify two kinds of constituents contained in Prunellae Spica. Principal component analysis (PCA) was applied to compare the similarity and difference among samples from different regions of China. In the present study, 22 compounds were identified and some new fragmental pathways of triterpenic acids were discovered. An accurate and reliable HPLC-ELSD method was developed and validated for the first time to simultaneously quantify multiple constituents, including rosmarinic acid, maslinic acid, corosolic acid, betulin, oleanolic acid, and ursolic acid in the extract of Prunellae Spica. (PCA) revealed some similarities and differences among different samples from different regions of China. In conclusion, our results from this study would be helpful in establishing a scientific and rational quality control method for Prunellae Spica.


Subject(s)
China , Chromatography, High Pressure Liquid , Methods , Cinnamates , Chemistry , Depsides , Chemistry , Drugs, Chinese Herbal , Chemistry , Molecular Structure , Prunella , Chemistry , Tandem Mass Spectrometry , Methods , Triterpenes , Chemistry
16.
Chinese Medical Journal ; (24): 1523-1528, 2015.
Article in English | WPRIM | ID: wpr-231744

ABSTRACT

<p><b>BACKGROUND</b>Previous studies have indicated that endoplasmic reticulum stress participates in and mediates liver injury and apoptosis in brain-dead (BD) rats. In this study, we observed the effect of salubrinal (Sal, Sigma, USA) on liver cells in BD rats and explored its relevant mechanisms.</p><p><b>METHODS</b>Thirty Sprague-Dawley rats were equally randomized into three groups: BD group, Sal group, and DMSO group. The BD models were established by increasing intracranial pressure in a modified, slow, and intermittent way. In the drug groups, Sal was administered 1 h before the induction of BD. After modeling was completed, the blood and liver samples were harvested. CHOP and Caspase-12 mRNA expression was detected using quantitative polymerase chain reaction. PKR-like ER kinase (PERK), P-eukaryotic translation initiation factor 2α (eIF2α), eIF2α, CHOP and caspase-12 expression was detected using western blotting (WB). CHOP and caspase-12 distribution and expression in liver tissues were determined using immunohistochemistry (IHC). Alanine aminotransferase and aspartate aminotransferase level were detected using an automatic biochemical analyzer. Hepatic cell apoptosis was detected using TUNEL. The results were analyzed using Quantity-one v4.62 software (Bio-Rad, USA).</p><p><b>RESULTS</b>CHOP and caspase-12 expression and PERK, eIF2α, and P-eIF2α protein expression showed no significant difference between BD group and DMSO group. Compared with BD group, Sal group had a significantly higher P-eIF2C level and a lower P-PERK level 2 h and 6 h after BD (P < 0.05). However, eIF2α expression showed no significant difference (P > 0.05). After the Sal treatment, CHOP and caspase-12 mRNA expression significantly decreased 4 h after BD (P < 0.05). WB and IHC indicated that CHOP and caspase-12 expression also significantly decreased after Sal treatment. Sal was associated with improved liver function and decreased hepatic cell apoptosis.</p><p><b>CONCLUSIONS</b>Sal can significantly reduce apoptosis in hepatic cells of BD rats. This protective effect may be achieved via the PERK-eIF2α signaling pathway.</p>


Subject(s)
Animals , Apoptosis , Blotting, Western , Brain Death , Metabolism , Caspase 12 , Genetics , Metabolism , Cinnamates , Disease Models, Animal , Endoplasmic Reticulum Stress , Eukaryotic Initiation Factor-2 , Genetics , Metabolism , Immunohistochemistry , Liver , Wounds and Injuries , Male , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Thiourea , Transcription Factor CHOP , Genetics , Metabolism
17.
Acta Pharmaceutica Sinica ; (12): 861-867, 2015.
Article in Chinese | WPRIM | ID: wpr-257055

ABSTRACT

Using bioactive compounds 7-hydroxy flavone, salicylaldehyde, cinnamic acid and 4-amino-5- mercapto-1,2,4-triazoles as starting materials, three new types of flavone derivatives containing 1,2,4-triazole structure were synthesized via different step reactions. These new compounds were characterized by 1IHNMR, ESI-MS, IR and elemental analysis. Their scavenging effects on the superoxide radical (O2·-), hydroxyl radical (·OH), DPPH · radical and their total reduction activities were tested. The results showed that all of the compounds possessed some antioxidative activity at the concentration of 0.5 mg · mL(-1), but the scavenging ability of the target compounds was lower than that of the standard compound Vc.


Subject(s)
Cinnamates , Chemistry , Flavones , Chemistry , Flavonoids , Chemistry , Free Radical Scavengers , Chemistry , Triazoles , Chemistry
18.
Article in Chinese | WPRIM | ID: wpr-330259

ABSTRACT

The difference between three representative components of total salvianolic acids in pharmacodynamic activity were compared by three different pharmacological experiments: HUVECs oxidative damage experiment, 4 items of blood coagulation in vitro experiment in rabbits and experimental myocardial ischemia in rats. And the effects of contribution rate of each component were calculated by multi index comprehensive evaluation method based on CRITIC weights. The contribution rates of salvianolic acid B, rosmarinic acid and Danshensu were 28.85%, 30.11%, 41.04%. Apparent oil/water partition coefficient of each representative components of total salvianolic acids in n-octyl alcohol-buffer was tested and the total salvianolic acid components were characterized based on a combination of the approach of self-defined weighting coefficient with effects of contribution rate. Apparent oil/water partition coefficient of total salvianolic acids was 0.32, 1.06, 0.89, 0.98, 0.90, 0.13, 0.02, 0.20, 0.56 when in octanol-water/pH 1.2 dilute hydrochloric acid solution/ pH 2.0, 2.5, 5.0, 5.8, 6.8, 7.4, 7.8 phosphate buffer solution. It provides a certain reference for the characterization of components.


Subject(s)
Animals , Benzofurans , Chemistry , Pharmacology , Cinnamates , Chemistry , Pharmacology , Depsides , Chemistry , Pharmacology , Lactates , Chemistry , Pharmacology , Male , Rabbits , Rats , Rats, Sprague-Dawley , Solubility
19.
Clinics ; 69(2): 134-144, 2/2014. graf
Article in English | LILACS | ID: lil-701378

ABSTRACT

OBJECTIVE: The present study aimed to investigate the mechanisms underlying the anti-inflammatory and anti-angiogenic effects of ethyl-p-methoxycinnamate isolated from Kaempferia galanga. METHODS: The anti-inflammatory effects of ethyl-p-methoxycinnamate were assessed using the cotton pellet granuloma assay in rats, whereby the levels of interleukin-1 and tumor necrosis factor-α were measured in the animals' blood. In addition, the levels of interleukin, tumor necrosis factor, and nitric oxide were measured in vitro using the human macrophage cell line (U937). The analgesic effects of ethyl-p-methoxycinnamate were assessed by the tail flick assay in rats. The anti-angiogenic effects were evaluated first by the rat aortic ring assay and, subsequently, by assessing the inhibitory effects of ethyl-p-methoxycinnamate on vascular endothelial growth factor, proliferation, migration, and tube formation in human umbilical vein endothelial cells. RESULTS: Ethyl-p-methoxycinnamate strongly inhibited granuloma tissue formation in rats. It prolonged the tail flick time in rats by more than two-fold compared with the control animals. The inhibition of interleukin and tumor necrosis factor by ethyl-p-methoxycinnamate was significant in both in vivo and in vitro models; however, only a moderate inhibition of nitric oxide was observed in macrophages. Furthermore, ethyl-p-methoxycinnamate considerably inhibited microvessel sprouting from the rat aorta. These mechanistic studies showed that ethyl-p-methoxycinnamate strongly inhibited the differentiation and migration of endothelial cells, which was further confirmed by the reduced level of vascular endothelial growth factor. CONCLUSION: Ethyl-p-methoxycinnamate exhibits significant anti-inflammatory potential by inhibiting pro-inflammatory cytokines and angiogenesis, thus inhibiting the main functions of endothelial cells. Thus, ethyl-p-methoxycinnamate could be a promising therapeutic agent ...


Subject(s)
Animals , Humans , Male , Rats , Angiogenesis Inhibitors/pharmacology , Anti-Inflammatory Agents/pharmacology , Cinnamates/pharmacology , Plant Extracts/pharmacology , Vascular Endothelial Growth Factor A/drug effects , Zingiberaceae/chemistry , Analysis of Variance , Angiogenesis Inhibitors/isolation & purification , Anti-Inflammatory Agents/isolation & purification , Cell Proliferation/drug effects , Enzyme-Linked Immunosorbent Assay , Human Umbilical Vein Endothelial Cells/drug effects , Interleukin-1/analysis , Rats, Sprague-Dawley , Reproducibility of Results , Tumor Necrosis Factor-alpha/analysis , Tumor Necrosis Factor-alpha/drug effects , /drug effects , Vascular Endothelial Growth Factor A/analysis
20.
Protein & Cell ; (12): 382-393, 2014.
Article in English | WPRIM | ID: wpr-757502

ABSTRACT

Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer (SCNT). Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treatment with various histone deacetylase inhibitors (HDACi), including Trichostatin A, Scriptaid, and m-carboxycinnamic acid bishydroxamide (CBHA), in which only the effect of CBHA has not yet been tested in other species. In this paper we examine the effect of CBHA treatment on the development of porcine SCNT embryos. We have discovered the optimum dosage and time for CBHA treatment: incubating SCNT embryos with 2 μmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 (AcH3K9), acetylation at histone 3 lysine 18 (AcH3K18), and acetylation at histone 4 lysine 16 (AcH4K16) was raised after CBHA treatment. Meanwhile, CBHA treatment improved the expression of development relating genes such as pou5f1, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was not significantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by CBHA treatment.


Subject(s)
Acetylation , Animals , Blastocyst , Cell Biology , Cell Nucleus , Metabolism , Cinnamates , Pharmacology , Embryo, Mammalian , Metabolism , Embryonic Development , Epigenesis, Genetic , Female , Gene Expression , Histone Deacetylase Inhibitors , Pharmacology , Histones , Metabolism , Homeodomain Proteins , Genetics , Metabolism , In Vitro Techniques , Insulin-Like Growth Factor II , Genetics , Metabolism , Nuclear Transfer Techniques , Octamer Transcription Factor-3 , Genetics , Metabolism , Swine
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