RESUMEN
Balanophora harlandii Hook (B. harlandii), a folk medicine, has been traditionally employed to treat traumatic bleeding, gastroenteritis, icteric hepatitis, hemorrhoids, and other conditions. In this work, polysaccharides with anti-inflammatory effects were extracted from B. harlandii and purified. The extraction conditions were optimized, and the properties of one purified neutral fraction, denoted as BHPs-W-S3, were analyzed. BHPs-W-S3 has a molecular weight of 14.1â¯kDa, and its three main monosaccharides are glucose, galactose, and xylose, with a molar ratio of 6.4:1.7:1.1. Its main chain consists of â6)-α-D-Glcp-(1â, â4,6)-α-D-Glcp-(1â, â6)-ß-D-Galp-(1â, â3,6)-ß-D-Galp-(1â, and it has branch chains at the O-4 and/or O-3 positions. In addition, in vitro experiments showed that the polysaccharides from B. harlandi can decrease the phosphorylation level of p65 and IκBα in LPS-induced RAW264.7 cells to reduce the expression of the pro-inflammatory genes such as TNF-α, IL-6, and IL-1ß.
Asunto(s)
Antiinflamatorios , Polisacáridos , Ratones , Animales , Polisacáridos/farmacología , Polisacáridos/química , Polisacáridos/aislamiento & purificación , Antiinflamatorios/farmacología , Antiinflamatorios/química , Antiinflamatorios/aislamiento & purificación , Células RAW 264.7 , Peso Molecular , Balanophoraceae/química , Extractos Vegetales/farmacología , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Lipopolisacáridos/farmacología , FN-kappa B/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Inflamación/tratamiento farmacológicoRESUMEN
An UHPLC/LC-MS was founded to detect balanophorin B (B), gallic acid (GA), 4-hydroxycinnamic acid (HC), and their in vivo profiling in rats, after oral administration of the ethanol extract of Balanophora simaoensis S. Y. Chang et Tam. The in vivo dynamic existence of 3 molecular entities in rats and the multistep biotransformation of GA were elucidated by their sensitive mass spectrometry response after efficient UHPLC and/or HPLC separation, through analyzing the bio-samples of rat plasma, bile, liver, kidneys, and excreta. The method was validated with satisfactory calibration curves having correlation coefficients r from 0.996 to 0.999 for concentration scaled from 0.100 nM to 0.100 µM, internal standard normalized matrix factors ranged from 0.923 to 0.993, sextuplicate recoveries valued from 95.0% to 103.6%, as well as accuracy and precision varied from 95.6% to 103.7%. The content of B, GA, and HC in the whole herb was of 4.66, 63.5, and 10.4 µmol/kg in dry weight, respectively. The Cmax for B, GA, and HC in rat systemic circulation was of 76.0 nM, 2.30 µM, and 51.0 µM, with tmax at 3, 2, and 2 h, respectively. B and GA stayed in rat liver over 4 hs to present a material base for the pharmacology and pharmacodynamics of the whole herb. The biotransformation of GA indicated a complicated scheme in rats. As a final metabolite from GA with total biotransformation conversion over 20%, 4-hydroxybenzaldehyde resourced from two steps of dehydroxylation and one step of reduction of GA, but not concerned with HC.
Asunto(s)
Balanophoraceae , Ácidos Cumáricos , Medicamentos Herbarios Chinos , Ácido Gálico , Animales , Masculino , Ratas , Administración Oral , Balanophoraceae/química , Cromatografía Líquida de Alta Presión/métodos , Ácidos Cumáricos/administración & dosificación , Ácidos Cumáricos/sangre , Ácidos Cumáricos/farmacocinética , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/análisis , Medicamentos Herbarios Chinos/farmacocinética , Ácido Gálico/administración & dosificación , Ácido Gálico/sangre , Ácido Gálico/farmacocinética , Espectrometría de Masas/métodos , Ratas Sprague-DawleyRESUMEN
Lophophytum species are holoparasites that grow on tree roots. The objectives of the work were to explore the chemical composition of the tubers of two Lophophytum species and to analyze the antioxidant, anti-inflammatory and antilithiatic activity of their extracts using in vitro methods. The chemical composition was determined by histochemical, phytochemical and TLC tests. In addition, the profile of phenolic compounds was determined by HPLC-MS. The presence of secondary metabolites of recognized activity was demonstrated. The results of the HPLC-MS/MS allowed the tentative identification of catechin, luteolin and glycosides of eriodictyol, naringenin and luteolin in the extract of Lophophytum leandriand eriodictyol, naringenin, luteolin and their glycosylated derivatives in Lophophytum mirabile. The extracts showed promising antioxidant (DPPH, ABTS and ß-carotene-linoleic acid), anti-inflammatory (inhibition of 5-LOX) and anti-urolytic (by bioautographic TLC) activity. It is noteworthy that these are the first results of the phytochemical composition and biological activity of L. mirabile. However, in vivo studies are required to corroborate these activities.
Las especies de Lophophytumson holoparásitas que crecen en raíces de árboles. Los objetivos del trabajo fueron explorar la composición química del túber de dos especies de Lophophytum y analizar la actividad antioxidante, antiinflamatoria y antilitiásica de sus extractos usando métodos in vitro. La composición química se determinó mediante pruebas histoquímicas, fitoquímicas y por TLC. Además, se determinó el perfil de compuestos fenólicos por HPLC-MS/MS. Se demostró presencia de metabolitos secundarios de reconocida actividad. Los resultados del HPLC-MS/MS permitieron identificar tentativamente catequina, luteolina y glucósidos de eriodictiol, naringenina y luteolina en el extracto de Lophophytum leandriy eriodictiol, naringenina, luteolina y sus derivados glicosilados en Lophophytum mirabile. Los extractos mostraron prometedora actividad antioxidante (DPPH, ABTS y ß-caroteno-ácido linoleico), antiinflammatoria (inhibición de la 5-LOX) y antiurolitiásica (por TLC bioautográfica). Es de destacar que estos son los primeros resultados de composición fitoquímica y actividad biológica de L. mirabile. Sin embargo, se requieren estudios in vivo para corroborar dichas actividades.
Asunto(s)
Extractos Vegetales/farmacología , Extractos Vegetales/química , Balanophoraceae/química , Cromatografía Líquida de Alta Presión , Flavanonas/análisis , Flavonas/análisis , Compuestos Fenólicos/análisis , Espectrometría de Masas en Tándem , Antiinflamatorios/farmacología , Antiinflamatorios/química , Antioxidantes/farmacología , Antioxidantes/químicaRESUMEN
Renal interstitial fibrosis (RIF) is a crucial pathological change leading to chronic kidney disease (CKD). Currently, no effective medicines have been available for treating it. In our research, we examined the effects of polysaccharides extracted from Balanophora polyandra Griff (BPPs) on kidney fibrosis and epithelial to mesenchymal transition (EMT) in vivo and in vitro, aiming to explore the underlying mechanisms. By using the mice with unilateral urethral obstruction (UUO) as experimental subjects, we examined the medicinal values of BPPs on alleviating RIF. The effects of BPPs were evaluated by examining the histological staining and relative mRNA and protein expression levels of the related genes. The possible underlying mechanisms were further explored with human normal renal proximal tubular epithelia (HK-2 cells) as in vitro model. In UUO mice, BPP treatment could significantly alleviate interstitial fibrosis through reducing the components (Collagens I, III and IV) of extracellular matrix (ECM), and reducing the activation of fibroblasts producing these components, as revealed by inhibiting the hallmarks (fibronectin and α-SMA) of fibroblast activation. Furthermore, BPP administration increased the expression levels of matrix metalloproteinases (MMPs) and declined those of tissue inhibitors of metalloproteinases (TIMPs). BPPs markedly ameliorated EMT in both the kidneys of UUO mice and TGF-ß1 treated HK-2 cells. Moreover, BPP treatment decreased the expression levels of several transcriptional factors involved in regulating E-cadherin expression, including snail, twist and ZEB1. Additionally, the Hedgehog pathway was found to be closely correlated with renal fibrosis and EMT. Altogether, our results clearly demonstrated that BPP treatment effectively inhibited the Hedgehog pathway both in renal tissues of UUO mice and TGF-ß1-treated HK-2 cells. Thus, BPPs ameliorated RIF and EMT in vivo and in vitro via suppressing Hedgehog signalling pathway.
Asunto(s)
Balanophoraceae/química , Transición Epitelial-Mesenquimal/efectos de los fármacos , Proteínas Hedgehog/metabolismo , Enfermedades Renales/metabolismo , Polisacáridos/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Línea Celular , Colágeno/metabolismo , Modelos Animales de Enfermedad , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Matriz Extracelular/metabolismo , Fibrosis , Humanos , Enfermedades Renales/tratamiento farmacológico , Enfermedades Renales/etiología , Enfermedades Renales/patología , Túbulos Renales Proximales , Masculino , Ratones , Polisacáridos/química , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Two new butenolides, balanolides A (1) and B (1), were isolated from the aerial parts of the parasitic plant Balanophora fungosa. Their structures were established by comprehensive spectroscopic analysis and quantum chemical ECD calculation. The new butenolides were evaluated for their inhibitory effects against nitric oxide (NO) production in LPS-induced RAW 264.7 macrophage cells. Compounds 1 and 2 displayed moderate anti-inflammatory activity with IC50values of 11.8 and 12.9 µM, respectively.
Asunto(s)
4-Butirolactona/análogos & derivados , Antiinflamatorios/farmacología , Balanophoraceae/química , 4-Butirolactona/farmacología , Animales , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Ratones , Óxido Nítrico/biosíntesis , Extractos Vegetales/química , Células RAW 264.7RESUMEN
AIMS: Cancer cells exhibit a metabolic change called aerobic glycolysis compared with normal cells. Balanophorin B is a terpenoid ingredient reported from the genus Balanophora. In this research, we studied the effect of balanophorin B on glycolysis of HepG2 cells and Huh-7 cells under hypoxia. MAIN METHODS: The Warburg effect was monitored by assessing glucose uptake, oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Key enzymes in the glycolytic pathway and HIF-1α protein expression and degradation were analyzed by real-time PCR and western blotting. The anti-cancer effect of balanophorin B in vivo was also investigated. KEY FINDINGS: Balanophorin B inhibited the proliferation, glucose uptake, and ECAR in both HepG2 cells and Huh-7 cells. In addition, balanophorin B inhibited the protein level of HIF-1α and its downstream targets LDHA and HKII under hypoxia, whereas HIF-1α mRNA level did not change after balanophorin B treatment. The HIF-1α plasmid reversed the inhibition of balanophorin B on glycolysis, and the proteasome inhibitor MG132 attenuated the effect of balanophorin B on HIF-1α protein expression, suggesting that balanophorin B might post-transcriptionally affect HIF-1α. Moreover, balanophorin B increased the expression of VHL and PHD2. HIF-1α siRNA also greatly attenuated the inhibitory effect of balanophorin B on HepG2 cells glucose uptake. Balanophorin B significantly inhibited tumor growth in vivo, without causing obvious toxicity to mice. SIGNIFICANCE: These data suggest that balanophorin B inhibits glycolysis probably via an HIF-1α-dependent pathway, and the ubiquitin-proteasome pathway was greatly involved in the induction of balanophorin B on HIF-1α degradation.
Asunto(s)
Glucólisis/efectos de los fármacos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Terpenos/farmacología , Animales , Balanophoraceae/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ciclo del Ácido Cítrico , Glucosa/metabolismo , Células Hep G2 , Humanos , Ratones , Ratones Desnudos , Extractos Vegetales/farmacología , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Balanophora involucrata J. D. Hooker has been known to possess potential anti-inflammatory and antibacterial activities; however, its antiviral activity has not been evaluated so far. In order to find new neuraminidase inhibitors (NAIs), the neuraminidase (NA) inhibition activity of different B. involucrata extracts was evaluated. In this study, an in vitro NA inhibition assay was performed to identify which extract of B. involucrata exhibits (maximal) inhibitory activity against NA. Ultra high performance liquid chromatography/quadrupole time-of-flight-tandem mass spectroscopy (MS/MS) and molecular docking techniques were used to identify the specific compounds responsible for the anti-influenza activity of the extract, and to explore the potential natural NAIs. The ethyl acetate extract of B. involucrata exhibited significant inhibitory activity against NA with 50% inhibitory concentration (IC50 ) value of 159.5 µg/mL. Twenty compounds were identified according to the MS/MS spectra; among them two compounds (quercitrin and phloridzin) showed obvious inhibitory activity against NA, with IC50 of 311.76 and 347.32 µmol/L, respectively. This study suggested that B. involucrata can be a potential natural source of NAIs and may be useful in the fight against ferocious influenza viruses.
Asunto(s)
Balanophoraceae/química , Inhibidores Enzimáticos , Neuraminidasa/antagonistas & inhibidores , Extractos Vegetales/química , Cromatografía Líquida de Alta Presión , Inhibidores Enzimáticos/análisis , Inhibidores Enzimáticos/aislamiento & purificación , Inhibidores Enzimáticos/farmacología , Flavonoides/análisis , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Simulación del Acoplamiento Molecular , Espectrometría de Masas en TándemRESUMEN
Three new muurolane type sesquiterpene glycosides, named balanoindicosides A - C (1 - 3), were isolated from the whole plant of Balanophora fungosa subsp. indica using various chromatographic methods. Their structures were determined by extensive analysis of HR-ESI-MS and NMR spectroscopy. The stereochemistry of muurolane sesquiterpene backbone was demonstrated by NOESY analysis. Configuration of C-12 in compounds 2 and 3 could be distinguished by chemical shift value of C-14. Compounds 1-3 exhibited weak cytotoxic activity towards HepG-2, HL-60, LU-1, and MCF-7 cell lines.
Asunto(s)
Balanophoraceae/química , Glicósidos/química , Sesquiterpenos/química , Sesquiterpenos/farmacología , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Glicósidos/farmacología , Células HL-60 , Células Hep G2 , Humanos , Células MCF-7 , Espectroscopía de Resonancia Magnética , Estructura Molecular , Sesquiterpenos Policíclicos/química , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Thonningia sanguinea is a parasitic herb widely used in traditional African medicine. Dihydrochalcone glucosides (unsubstituted, substituted with hexahydroxydiphenoyl or galloyl moieties) are the main constituents in the subaerial parts of this plant. In the present study, purification of the six major compounds from a methanol extract of the plant's subaerial parts was achieved by centrifugal partition chromatography. A first dimension centrifugal partition chromatography separation with the solvent system methyl tert-butyl ether/1,2-dimethoxyethane/water (1:2:1) in the ascending mode enabled the isolation of the two major bioactive compounds thonningianin A and B from 350 mg of methanol extract within only 16 min with respectable yields (25.7 and 21.1 mg), purities (87.1 and 85%), and recoveries (71.2 and 70.4%). Using a multiple heart-cutting strategy, the remaining four major dihydrochalcone glucosides of the extract were further separated in a second dimension centrifugal partition chromatography with the solvent system ethyl acetate/1,2-dimethoxyethane/water (2:1:1) in the descending mode with high purities (88.9-98.8%).
Asunto(s)
Balanophoraceae/química , Taninos Hidrolizables/aislamiento & purificación , Extractos Vegetales/aislamiento & purificación , Centrifugación , Distribución en Contracorriente , Taninos Hidrolizables/química , Medicinas Tradicionales Africanas , Extractos Vegetales/químicaRESUMEN
Thonnigia sanguinea is a plant widely used in traditional African medicine against a variety of diseases. The obligate parasite is growing throughout tropical African forests and utilizes a large variety of hosts. Dihydrochalcone glucoside derivatives isolated from the subaerial parts of this plant were identified as potential antidiabetic lead compounds. In this study, an ultrahigh-performance liquid chromatographic method coupled with a photodiode array detector was developed for the quantitation of six major dihydrochalcone derivatives. The analytes were baseline separated in complex samples within 14 minutes on a Phenomenex Luna Omega 1.6 µm C18 column using a mobile phase consisting of water and acetonitrile (each + 0.01% trifluoroacetic acid) in gradient elution. Method validation confirmed the selectivity, linearity (R2 ≥ 0.9992), precision (inter-day ≤ 1.98%, intraday ≤ 2.00%), and accuracy (recovery rates of 97.4â-â106.3% for all analytes). At 280 nm, the LODs and LOQs were found to be lower than 1.42 and 4.30 µg/mL, respectively. Eight plant batches from the northern Angolan province of Uíge (collected in the wild or bought on markets) were extracted with methanol using an ultrasound-assisted extraction protocol and subsequently analyzed with the validated method. Results indicated high contents of dihydrochalcone glucosides in all eight samples. Most notably, the two bioactive constituents thonningianin A and B were present in fairly large amounts (2.42â-â5.35 w%).
Asunto(s)
Balanophoraceae/química , Chalconas/análisis , Cromatografía Líquida de Alta Presión/métodos , Glucósidos/análisis , Dispositivos Laboratorio en un Chip , Procedimientos Analíticos en Microchip/métodosRESUMEN
Six new and four known dihydrochalcone glucoside derivatives (1-10), the phenylpropanoid coniferin (11), and the lignans (+)-pinoresinol (12) and lariciresinol (13) were isolated from the subaerial plant parts of Thonningia sanguinea in the course of a screening campaign for new antidiabetic lead compounds. The structures of the new substances were elucidated by HRESIMS, NMR, GC-MS, and ECD data evaluation. 2'- O-(3-Galloyl-4,6- O- Sa-hexahydroxydiphenoyl-ß-d-glucopyranosyl)-3-hydroxyphloretin (4), 2'- O-(4,6- O- Sa-hexahydroxydiphenoyl-ß-d-glucopyranosyl)phloretin (5), 2'- O-(3- O-galloyl-4,6- O- Sa-hexahydroxydiphenoyl-ß-d-glucopyranosyl)phloretin (6), and thonningianin B (9) showed moderate protein tyrosine phosphatase-1B inhibition in an enzyme assay (IC50 values ranging from 19 to 25 µM), whereas thonningianin A (10) was identified as a more potent inhibitor (IC50 = 4.4 µM). The observed activity differences could be explained by molecular docking experiments. The activity of 10 could further be confirmed in HEPG2 liver carcinoma cells, where the compound was able to increase the level of phosphorylated insulin receptors in a concentration-dependent manner.
Asunto(s)
Balanophoraceae/química , Chalconas/aislamiento & purificación , Glucósidos/aislamiento & purificación , Proteína Tirosina Fosfatasa no Receptora Tipo 1/antagonistas & inhibidores , Chalconas/química , Chalconas/farmacología , Cromatografía de Gases y Espectrometría de Masas , Glucósidos/química , Glucósidos/farmacología , Células Hep G2 , Humanos , Espectroscopía de Resonancia Magnética , Simulación del Acoplamiento Molecular , Extractos Vegetales/farmacologíaRESUMEN
The bioactive substance investigation of Balanophora involucrata obtained 15 phenolic acids, including 5 new compounds (1-3, 8, 9), which were determined by various spectroscopic data analyses. Most isolated compounds displayed inhibitory effects on α-Glucosidase in vitro. For the potential inhibitors 8 (1.95µM) and 10 (9.02µM), the inhibition kinetics have been studied, which gave the Ki values as 0.68, 3.15µM respectively. And, in silico docking analyses have been performed to investigate the inhibitory mechanism of compounds 8 and 10. Additionally, most isolated compounds showed anti-oxidant activities in the DPPH scavenging assay. New compound 8 also could inhibit the acetyl transfer activity of GlmU moderately with the IC50 value of 18.21µM, which was a new antibacterial target.
Asunto(s)
Balanophoraceae/química , Depuradores de Radicales Libres/química , Inhibidores de Glicósido Hidrolasas/química , Hidroxibenzoatos/química , Depuradores de Radicales Libres/aislamiento & purificación , Inhibidores de Glicósido Hidrolasas/aislamiento & purificación , Hidroxibenzoatos/aislamiento & purificación , Simulación del Acoplamiento Molecular , Extractos Vegetales/químicaRESUMEN
Balanophora involucrata HOOK.f. & THOMSON (Balanophoraceae) is a parasite plant often growing on the roots of leguminous plants. The whole herb has been used medicinally for the treatment of irregular menstruation, cough, hemoptysis, traumatic injury and bleeding, dizziness and gastralgia in Yunnan Province, China. The 2,2-diphenyl-2-picrylhydrazyl (DPPH) assay on the 60% aq. acetone extract of the fresh whole plant of B. involucrata showed considerable radical-scavenging activity (SC50 15.3 µg/ml). Further purification on the extract led to the isolation of one new phenolic glycoside, sieboldin-3'-ketocarboxylic acid (1), and one new cyanogenic glycoside, proacacipetalin 6'-O-ß-D-glucopyranoside (2), together with 26 known compounds including three 4"-O-galloyl and 2",3"-O-(S)-hexahydroxydiphenoyl (HHDP) derivatives of dihydrochalcone glucosides, seven hydrolyzable tannins, and alkane glycosides. The cyanogenic compound isolated from the Balanophoraceae family for the first time might be a signal molecule between B. involucrata and its hosts. The free-radical-scavenging activity of the isolated compounds was also examined by DPPH assay.
Asunto(s)
Balanophoraceae/química , Depuradores de Radicales Libres/química , Glicósidos/química , Fenoles/química , Depuradores de Radicales Libres/aislamiento & purificación , Glicósidos/aislamiento & purificación , Raíces de Plantas/químicaRESUMEN
One new flavanone (S)-5,7,3',5'-tetrahydroxy-flavanone-7-O-(6â³-galloyl)-ß-D-glucopyranose (1), together with one known flavanone and four known dihydrochalcones were isolated from the ethyl acetate partitions from an alcoholic extract of the whole plant of Balanophora involucrata Hook. f. All of the compounds were characterized by spectroscopic methods. Their in vitro BACE inhibitory effects were evaluated. Compounds 1, 2, 4, and 5 were found to be a little more active than the positive control, although all of which are below 50% inhibition at 10µM.
Asunto(s)
Balanophoraceae/química , Flavanonas/farmacología , Chalconas/química , Flavanonas/química , Estructura MolecularRESUMEN
A bioassay-oriented study was carried-out in order to validate the traditional uses of Lophophytum leandri, a parasitic plant used as a diuretic in traditional medical practices of Argentina. Four known flavonoids have been isolated from the active fraction. Quercetin-3-O-beta-D-glucopyranoside was identified as the active principle of the fraction. However, the diuretic activity of the extract and of the most active fraction had greater activity than that of the pure isolated compounds.
Asunto(s)
Balanophoraceae , Diuréticos/farmacología , Extractos Vegetales/farmacología , Animales , Balanophoraceae/química , Masculino , Ratas , Ratas WistarRESUMEN
OBJECTIVE: MicroRNAs (miRNAs) play important roles in cell proliferation, differentiation and apoptosis. 1, 3, 4-tri-O-galloyl-6-O-caffeoyl-ß-D-glucopyranose (BJA32515) is a new natural ellagitannin compound extracted from Balanophora Japonica MAKINO. The effect of BJA32515 on the expression of miRNAs in cancer cells has not yet been explored. Objective The present study was carried out to examine the changes in miRNA expression profiles in human HepG(2) hepatocarcinoma cells following BJA32515 exposure. METHODS: The proliferation of BJA32515-exposed HepG(2) cells was assessed using a colorimetric assay (cell counting kit-8). The miRNA expression profile of the cancer cells was analyzed using a miRNA array and quantitative real-time PCR. Apoptosis was assessed by annexin V and propidium iodide staining. RESULTS: BJA32515 inhibited the cell proliferation and increased apoptosis in HepG(2) cancer cells. The exposure to BJA32515 also caused alterations in the miRNA expression profile in the cells, with 33 miRNAs upregulated and 59 down-regulated. The up-regulation of let-7a and miR-29a and the down-regulation of miR-373 and miR-197 were verified by quantitative real-time PCR. CONCLSION: BJA32515-modifed miRNA expression may mediate the antiproliferative effect of this compound in HepG(2) cancer cells.
Asunto(s)
Apoptosis/efectos de los fármacos , Balanophoraceae/química , Ácidos Cafeicos/farmacología , Glucósidos/farmacología , Taninos Hidrolizables/farmacología , MicroARNs/metabolismo , Antineoplásicos/farmacología , Ácidos Cafeicos/aislamiento & purificación , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Glucósidos/aislamiento & purificación , Células Hep G2 , Humanos , Taninos Hidrolizables/aislamiento & purificación , MicroARNs/genética , PolifenolesRESUMEN
Tyrosinase, trypsin, and tryptase are known to play important roles in melanin production of human skin. This paper describes the study of the inhibitory effect of Balanophora fungosa on melanin. The 50% EtOH extract obtained from B. fungosa indicated an inhibitory effect on mushroom tyrosinase activity with an IC(50) value of 15 µg/mL. Bioassay-guided fractionation of the active extract resulted in the isolation of four known compounds. Their structures were identified as 1-O-(E)-caffeoyl-3-O-galloyl-4,6-(S)-HHDP-ß-d-glucopyranose (1), 1-O-(E)-caffeoyl-3,4,6-tri-O-galloyl-ß-d-glucopyranose (2), caffeoyl-ß-d-glucopyranose (3), and abietin (4) on the basis of spectroscopic analyses and comparison of their spectral data with those in the literature. Compounds 1 and 2 prevented pigmentation of melanin in a three-dimensional cultured human skin model. Furthermore, compounds 1 and 2 indicated inhibitory activities against trypsin and tryptase.
Asunto(s)
Balanophoraceae/química , Melaninas/antagonistas & inhibidores , Melaninas/biosíntesis , Extractos Vegetales/farmacología , Línea Celular Tumoral , Inhibidores Enzimáticos/farmacología , Humanos , Melanocitos/metabolismo , Melanoma , Modelos Biológicos , Monofenol Monooxigenasa/antagonistas & inhibidores , Extractos Vegetales/química , Piel , Inhibidores de Tripsina/farmacología , Triptasas/antagonistas & inhibidoresRESUMEN
The high polysaccharide content of some plant species hinders the successful isolation of their DNA. As an alternative to the macro-extraction methods previously published for polysaccharide-rich plants, we present two techniques (STE/CTAB and HEPES/CTAB), which are performed in microcentrifuge tubes. These protocols are suitable for small amounts of silica gel-preserved plant tissue such as are commonly available from endangered plants. The critical step to remove polysaccharides was performing initial washes in either STE (0.25 M sucrose, 0.03 M Tris, 0.05 M EDTA) or HEPES (2% ß-mercaptoethanol, 0.2% PVP, 0.1 M HEPES, pH 8.0) buffer. Precipitating the DNA at room temperature with isopropanol also aided in decreasing polysaccharide co-precipitation. Of the two protocols we present the STE/CTAB method has the advantages of being more cost-effective and avoiding the use of the hazardous chemical ß-mercaptoethanol.
Asunto(s)
ADN de Plantas/aislamiento & purificación , Plantas/química , Polisacáridos/metabolismo , Balanophoraceae/química , Balanophoraceae/genética , Tampones (Química) , Cetrimonio , Compuestos de Cetrimonio/química , Técnicas Genéticas , Malvaceae/química , Malvaceae/genética , Hojas de la Planta/química , Hojas de la Planta/genética , Plantas/genética , Gel de Sílice , Tensoactivos/químicaRESUMEN
Five new phenylpropanoids, named balajaponins A-E (1-5), were isolated from Balanophora japonica, along with 24 known compounds. Among them, three hydrolysable tannins (6-8) showed specific in vitro α-glucosidase inhibition, with IC50 values in the range of 1-4 µM. Kinetic analysis revealed that they all acted in a noncompetitive mode.
Asunto(s)
Balanophoraceae/química , Inhibidores de Glicósido Hidrolasas , Taninos Hidrolizables/farmacología , Fenilpropionatos/farmacología , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Taninos Hidrolizables/química , Concentración 50 Inhibidora , Cinética , Espectroscopía de Resonancia Magnética , Estructura Molecular , Fenilpropionatos/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , alfa-Glucosidasas/metabolismoRESUMEN
OBJECTIVE: To study the effect of 1,2,6-Tri-O-galloyl-beta-D-glucopyranose (BJA32531) on the miRNA expression during BJA32531-induced cytotoxicity in human HepG2 hepatocarcinoma cells. METHODS: Cell proliferation was assessed using a colorimetric assay (cell counting kit-8). Apoptosis was assessed by annexin V and propidium iodide staining. The miRNA expression profile of the cancer cells was analyzed by a miRNA array and quantitative real-time PCR. RESULTS: BJA32531 inhibited the cell proliferation and increased apoptosis in HepG2 cancer cells. Cellular exposure to BJA32531 influenced the miRNA expression pattern in the cells, including 19 upregulated and 85 down-regulated miRNAs in the cells. The up-regulations of let-7a and miR-10b as well as the down-regulations of miR-132 and miR-125b were verified to be consistent with the the results of the miRNA array. CONCLUSION: Our study suggests that the mechanisms by which BJA32531 exerted the antiproliferative effects on HepG2 cancer cells may be related to its regulation of miRNA.