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We report the chemical composition of the crude leaf extracts obtained from Stizophyllum perforatum (Cham.) Miers (Bignoniaceae), a simple high-performance liquid chromatography-diode array detection (HPLC-DAD) method based on mangiferin as an internal standard to quantify verbascoside, and the verbascoside acute oral toxicity and antileishmanial activity. HPLC-high-resolution mass spectrometry-DAD (HPLC-HRMS-DAD) analyses of the crude ethanol S. perforatum leaf extracts (CE-1 and CE-2) revealed that verbascoside was the major constituent in both extracts. CE-1 was purified, and verbascoside and casticin, among other compounds, were isolated. The developed HPLC-DAD method was validated and met the required standards. Investigation of the CE-2 acute toxicity indicated a lethal dose (LD50) greater than 2,000 mg/kg of body weight. Both CE-1 and CE-2 exhibited antileishmanial activity. The isolated compounds, verbascoside and casticin, also displayed antileishmanial activity with effective concentrations (IC50) of 6.23 and 24.20 µM against promastigote forms and 3.71 and 18.97 µM against amastigote forms of Leishmania amazonensis, respectively, but they were not cytotoxic to J774A.1 macrophages. Scanning electron microscopy of the L. amazonensis promastigotes showed that the parasites became more rounded and that their plasma membrane was altered in the presence of verbascoside. Additionally, transmission electron microscopy demonstrated that vacuoles emerged, lipids accumulated, kinetoplast size increased, and interstitial extravasation occurred in L. amazonensis promastigotes exposed to verbascoside. These findings suggest that S. perforatum is a promising candidate for further in vivo investigations against L. amazonensis.
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MAIN CONCLUSION: SA and H2O2, in single and mixed elicitation stimulate specialized metabolism and activate oxidative stress in C. tenuiflora plants. Single elicitation with salicylic acid (SA at 75 µM) and, hydrogen peroxide (at 150 µM), and mixed elicitation (75 µM SA + 150 µM H2O2) were evaluated on specialized metabolism in Castilleja tenuiflora Benth. plants. Total phenolic content (TPC), phenylalanine ammonia-lyase (PAL) activity, antioxidant enzymes and specialized metabolite profiles, as well as the expression levels of eight genes involved in phenolic (Cte-TyrDC, Cte-GOT2, Cte-ADD, Cte-AO3, Cte-PAL1, Cte-CHS1) and terpene pathways (Cte-DXS1 and Cte-G10H) and their correlation with major metabolite (verbascoside and aucubin) concentrations were investigated. TPC content (three-fold) and PAL activity (11.5-fold) increased with mixed elicitation, as well as catalase and peroxidase activity (11.3-fold and 10.8-fold, respectively), compared to single elicitation. Phenylethanoid accumulation was greatest under mixed elicitation, followed by SA and H2O2. Lignan accumulation was differential, depending on the plant part and the elicitor. Flavonoids only appeared after mixed elicitation. The high concentration of verbascoside under mixed elicitation was related to a high gene expression. Single elicitation induced iridoid accumulation in specific parts (H2O2 in aerial parts and SA in roots), whereas under mixed elicitation, it accumulated in both parts. A high concentration of aucubin in the aerial part was related to a high expression level of genes of the terpene pathway Cte-DXS1 and Cte-G10H, and in the root with Cte-G10H, while Cte-DXS1 was downregulated in this tissue in all treatments. Mixed elicitation with SA and H2O2 represents an interesting tool to increase the production of specialized metabolites in plants.
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Peróxido de Hidrogênio , Orobanchaceae , Peróxido de Hidrogênio/metabolismo , Ácido Salicílico/metabolismo , Iridoides , Fenóis/metabolismo , Antioxidantes/metabolismo , Orobanchaceae/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Aloysia gratissima leaves are popularly used to treat respiratory, digestive, and nervous system disorders. Several studies have been carried out to determine the biological activity of A. gratissima, such as its antibacterial and anti-edematogenic activities, but despite the beneficial uses of A. gratissima, few studies have examined the toxicological profile of this plant. AIM OF THE STUDY: This study aimed to determine the chemical composition, cytotoxic, genotoxic, mutagenic potential, and antioxidant activity of an aqueous extract of A. gratissima leaves (AG-AEL). MATERIAL AND METHODS: The phytochemical constitution of AG-AEL was assessed by colorimetric analyses and High-performance liquid chromatography (HPLC). The inorganic elements were detected by Particle-Induced X-ray Emission (PIXE). The antioxidant, cytotoxicity, genotoxic, and mutagenic activities were evaluated in vitro by Di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH), Sulforhodamine B (SRB) assay, comet assay, and Salmonella/microsome assays. RESULTS: AG-AEL indicated the presence of terpenoids, flavonoids, and phenolic acids. HPLC detected rutin at 2.41 ± 0.33 mg/100 mg. PIXE analysis indicated the presence of Mg, Si, P, S, K, Ca, Mn, and Zn. The 50% inhibitory concentration was 84.17 ± 3.17 µg/mL in the DPPH assay. Genotoxic effects were observed using the Comet assay in neuroblastoma (SH-SY5Y) cells and mutations were observed in TA102 and TA97a strains. The extract showed cytotoxic activities against ovarian (OVCAR-3), glioblastoma (U87MG), and colon (HT-29) cancer cell lines. CONCLUSIONS: In conclusion, AG-AEL increased DNA damage, induced frameshift, and oxidative mutations, and showed cytotoxic activities against different cancer cells. The in vitro toxicological effects observed suggest that this plant preparation should be used with caution, despite its pharmacological potential.
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Neuroblastoma , Neoplasias Ovarianas , Humanos , Feminino , Apoptose , Extratos Vegetais/toxicidade , Extratos Vegetais/química , Linhagem Celular Tumoral , Mutagênicos/farmacologia , Antioxidantes/toxicidadeRESUMO
BACKGROUND: Pyrostegia venusta (Ker Gawl.) Miers occurs in threatened biodiversity hotspots of Cerrado and Atlantic forest biomes in Brazil and is used in traditional medicine to treat various respiratory and skin diseases. METHODS AND RESULTS: This study (i) examined the genetic diversity and structure of six natural populations of P. venusta from different Brazilian regions using sequence-related amplified polymorphism (SRAP) markers; and (ii) compared the intra- and inter-populational levels of the bioactive component verbascoside using high-performance liquid chromatography. The population from Nova Mutum, Mato Grosso, presented the highest genetic variability (Nei index H = 0.2759; Shannon index I = 0.4170; 85.14% polymorphic loci), whereas the population from Araxá, Minas Gerais, presented the lowest genetic variability (H = 0.1811; I = 0.2820; 70.27% polymorphic loci). The intra-populational variability (79%) was significantly higher (p = 0.001) than the inter-populational variability (21%). The populations were clustered into two groups but their genetic differentiation was not associated with geographical origin (Mantel test, r = 0.328; p > 0.05). The verbascoside content significantly differed (p > 0.05) among the six populations and between the individuals from each population. The highest verbascoside levels (> 200 µg/mg extract) were detected in populations from Araxá and Serrana, while the lowest verbacoside levels were detected in populations from Paranaíta and Sinop. CONCLUSIONS: This is the first report on the use of SRAP markers to analyze genetic variability in the family Bignoniaceae. Our findings shall help to better understand the genetic and chemical diversity of P. venusta populations, as well as provide useful information to select the most appropriate individuals to prepare phytomedicines.
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Bignoniaceae , Bignoniaceae/química , Bignoniaceae/genética , Variação Genética , Glucosídeos , Fenóis , PolifenóisRESUMO
Chemical profiling of Buddleja globosa was performed by high-performance liquid chromatography coupled to electrospray ionization (HPLC-DAD-ESI-IT/MS) and quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF/MS). The identification of 17 main phenolic compounds in B. globosa leaf extracts was achieved. Along with caffeoyl glucoside isomers, caffeoylshikimic acid and several verbascoside derivatives (ß-hydroxyverbascoside and ß-hydroxyisoverbascoside) were identified. Among flavonoid compounds, the presence of 6-hydroxyluteolin-7-O-glucoside, quercetin-3-O-glucoside, luteolin 7-O-glucoside, apigenin 7-O-glucoside was confirmed. Campneoside I, forsythoside B, lipedoside A and forsythoside A were identified along with verbascoside, isoverbascoside, eukovoside and martynoside. The isolation of two bioactive phenolic compounds verbascoside and forsythoside B from Buddleja globosa (Buddlejaceae) was successfully achieved by centrifugal partition chromatography (CPC). Both compounds were obtained in one-step using optimized CPC methodology with the two-phase solvent system comprising ethyl acetate-n-butanol-ethanol-water (0.25:0.75:0.1:1, v/v). Additionally, eight Natural Deep Eutectic Solvents (NADESs) were tested for the extraction of polyphenols and compared with 80% methanol. The contents of verbascoside and luteolin 7-O-glucoside after extraction with 80% methanol were 26.165 and 3.206 mg/g, respectively. Among the NADESs tested in this study, proline- citric acid (1:1) and choline chloride-1, 2- propanediol (1:2) were the most promising solvents. With these NADES, extraction yields for verbascoside and luteolin 7-O-glucoside were 51.045 and 4.387 mg/g, respectively. Taken together, the results of this study confirm that CPC enabled the fast isolation of bioactive polyphenols from B. globosa. NADESs displayed higher extraction efficiency of phenolic and therefore could be used as an ecofriendly alternative to classic organic solvents.
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Buddleja/química , Extratos Vegetais/química , Polifenóis/química , Centrifugação , Cromatografia , Extratos Vegetais/classificação , Extratos Vegetais/isolamento & purificação , Folhas de Planta/química , Polifenóis/classificação , Polifenóis/isolamento & purificação , Solventes/químicaRESUMO
The research on compounds exhibiting photoprotection against ultraviolet radiation (UVR) is a matter of increasing interest. The methanolic extract of a cell culture of Buddleja cordata has potential photoprotective effects as these cells produce phenolic secondary metabolites (SMs). These metabolites are attributed with biological activities capable of counteracting the harmful effects caused by UVR on skin. In the present work, the methanolic extract (310-2500 µg/mL) of B. cordata cell culture showed a photoprotective effect on UVB-irradiated 3T3-Swiss albino fibroblasts with a significant increase in cell viability. The greatest photoprotective effect (75%) of the extract was observed at 2500 µg/mL, which was statistically comparable with that of 250 µg/mL verbascoside, used as positive control. In addition, concentrations of the extract higher than 2500 µg/mL resulted in decreased cell viability (≤83%) after 24 h of exposure. Phytochemical analysis of the extract allowed us to determine that it was characterized by high concentrations of total phenol and total phenolic acid contents (138 ± 4.7 mg gallic acid equivalents and 44.01 ± 1.33 mg verbascoside equivalents per gram of extract, respectively) as well as absorption of UV light (first and second bands peaking at 294 and 330 nm, respectively). Some phenylethanoid glycosides were identified from the extract.
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Previously, our group reported the establishment of a white callus cell line of Buddleja cordata Kunth that is a high producer of the secondary metabolite, verbascoside (VB, also named acteoside), under suspension culture conditions. Here, we present experimental evidence of the sustained ability of that cellular line to grow and produce high amounts of VB for 5 years of continuous culture. Cellular line profiles were determined at the early (at the beginning) and late stages (at the end of 5 years of continuous subculturing) by analyzing relevant parameters of culture growth, i.e., specific growth rate [µ], doubling time [dt], and growth index [GI], as well as VB production. Late-stage cultures exhibited a 61% faster growth rate than early-stage subcultures, and 25 and 3% lower doubling time and growth index. The extents of growth phases were found to be different. Similar amounts of biomass were found (9.5 g and 9.4 g L-1). Verbascoside production increased parallel to cell growth; maximal yield level occurred in the mid-exponential phase and lasted until the end of the stationary phase (i.e., from the 15th to the 25th day and from the 9th to the 21st day for the early and late stages, correspondingly). The content of VB was higher in the late-stage culture (1.43 ± 0945 g L-1) than in the early-stage culture (1.21 ± 0.0286 g L-1). Productivity values point out the potential use of B. cordata cell line in the biotechnological production of VB and for research focused on the biochemistry of secondary metabolism.
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Stress is an important factor in the etiology of some illnesses such as gastric ulcers and depression. Castilleja tenuiflora Benth. (Orobanchaceae) is used in Mexican traditional medicine for the treatment of gastrointestinal diseases and nervous disorders. Previous studies indicated that organic extracts from C. tenuiflora had gastroprotective effects and antidepressant activity. In this study, we aimed to evaluate the gastroprotective and antidepressant activity of fractions and isolated compounds from the methanolic extract (MECt) of C. tenuiflora in stressed mice. Chromatographic fractionation of MECt produced four fractions (FCt-1, FCt-2, CFt-3, and FCt-4) as well as four bioactive compounds which were identified using TLC, HPLC and NMR analyses. The cold restraint stress (CRS)-induced gastric ulcer model followed by the tail suspension test and the forced swim test were used to evaluate the gastroprotective effect and antidepressant activity of the extract fractions. FCt-2 and FCt-3 at 100 mg/kg had significant gastroprotective and antidepressant effects. All isolated compounds (verbascoside, teniufloroside and mixture geniposide/ musseanoside) displayed gastroprotective effects and antidepressant activity at 1 or 2 mg/kg. The above results allow us to conclude that these polyphenols and iridoids from C. tenuiflora are responsible for the gastroprotective and antidepressant effects.
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Iridoides/uso terapêutico , Orobanchaceae/química , Extratos Vegetais/uso terapêutico , Animais , Antiulcerosos/química , Antiulcerosos/uso terapêutico , Antidepressivos/química , Antidepressivos/uso terapêutico , Cromatografia Líquida de Alta Pressão , Glucosídeos/química , Iridoides/química , Masculino , Metanol/química , Camundongos , Fenóis/química , Extratos Vegetais/química , Úlcera Gástrica/tratamento farmacológicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Verbena montevidensis and Verbena litoralis are plants that present morphological similarities. They are both known as "gervão" and "fel-da-terra", among other popular names, and are used in folk medicine to treat diseases related to the liver and stomach. AIMS OF THE STUDY: The aim of the current investigation was to determine the chemical composition and evaluate the hepatoprotective properties and cytotoxicity of the methanolic and aqueous extracts of V. montevidensis, V. litoralis and their main iridoid in HepG2 cells. MATERIALS AND METHODS: Aqueous and methanolic extracts from the dried aerial parts of V. montevidensis and V. litoralis were obtained. The methanolic extract of V. montevidensis afforded an iridoid as the main compound. The extracts and isolated compound were examined for the hepatoprotective effect and cytotoxicity in human hepatoblastoma HepG2 cells by MTT reduction and neutral red uptake methods. RESULTS: The methanolic and aqueous extracts of both species showed the presence of iridoid and phenylethanoids as the main compounds. The iridoid brasoside was isolated and identified by spectroscopic methods. The phenylethanoid was characterized by HPLC, comparing the UV profile and retention time with an authentic sample. The results of the biological assays indicate that both aqueous and methanolic extracts of V. montevidensis and V. litoralis as well as brasoside were hepatoprotective against ethanol-induced damage in HepG2 cells. The effect can be attributed to the main compounds present since both classes are recognized for this activity. CONCLUSIONS: Our results contribute towards validation of the traditional use of V. montevidensis and V. litoralis in the treatment of liver disorders.
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Glicosídeos/farmacologia , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Verbena , Sobrevivência Celular/efeitos dos fármacos , Células Hep G2 , Humanos , Hepatopatias/tratamento farmacológicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Plantago australis is a perennial plant widely distributed in Latin America, and its seeds and leaves are used in folk medicine to treat many diseases and conditions. Among its various chemical compounds, verbascoside is one of the most present, and has several pharmacological activities described, but there is not much information about its toxicity. AIMS OF THE STUDY: The aims of this study were to optimize the extraction of verbascoside from P. australis leaves with ultrasound methods, to develop a validated HPLC method to quantify verbascoside, and to evaluate the toxicological safety of the extract and verbascoside using in vitro and in vivo assays. MATERIALS AND METHODS: Dried leaves of P. australis were submitted to different extraction methods (percolation and ultrasound). The optimization of the ultrasound extraction was carried out by complete factorial design (22) and response surface methodology (RSM), followed by HPLC analysis for marker compounds. HPLC analysis was performed to verify the presence of the marker compounds aucubin, baicalein, oleanolic acid, ursolic acid and verbascoside. Mutagenicity was assessed by Salmonella/microsome mutagenicity assay. Cytotoxicity and genotoxicity were evaluated in V79 cells by reduction of tetrazolium salt (MTT) and neutral red uptake (NRU) assays, and alkaline comet assay, respectively. Verbascoside phototoxicity was assessed in 3T3 cells by the NRU phototoxicity assay. Wistar rats were used to perform the acute and sub-chronic toxicity tests. RESULTS: Among the marker compounds, only verbascoside was found in the hydroethanolic extract of P. australis leaves (PAHE); its highest concentration was obtained with the ultrasound-assisted extraction (UAE) method, optimized in 40â¯min and 25⯰C, and the method validation was successfully applied. Neither PAHE nor verbascoside showed mutagenic or genotoxic activities. Cytotoxicity assays demonstrated that both PAHE and verbascoside reduced cell viability only at the highest concentrations, and verbascoside had no phototoxic properties. The in vivo toxicity evaluation of PAHE suggested that the LD50 is higher than 5000â¯mg/Kg, indicating that this extract is safe for use. In addition, no signs of toxicity were found in subchronic exposure. CONCLUSION: The HPLC method to quantify verbascoside was validated, and the extraction of verbascoside from P. australis leaves through ultrasound method was optimized, yielding an extract with 6% verbascoside. Our results suggest the toxicological safety of PAHE and verbascoside, corroborating the use of P. australis in folk medicine, and also indicate verbascoside as a potential ingredient in topical formulations.
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Glucosídeos/toxicidade , Fenóis/toxicidade , Extratos Vegetais/toxicidade , Plantago , Células 3T3 , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cricetulus , Camundongos , Folhas de Planta , Ratos Wistar , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Testes de Toxicidade Aguda , Testes de Toxicidade SubcrônicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Because of the recent increase in type 2 diabetes and the need for complementary treatments in remote communities in many parts of the world, we undertook a study of treatments for diabetic symptoms used by traditional Q'eqchi' Maya healers of Belize. We used quantitative ethnobotany to rank culturally important taxa and subsequent pharmacological and phytochemical studies to assess bioactivity. MATERIALS AND METHODS: Antidiabetic plants identified in field interviews with traditional healers were ranked by syndromic importance value (SIV) based on 15 symptoms of diabetes. Species ranked with high SIV were tested in an assay relevant to many diabetes complications, the advanced glycation endproduct (AGE) inhibition assay. Active principles were identified by phytochemical analysis and bioassay. RESULTS: We collected over 70 plant species having a promising SIV score. The plants represented a broad range of neotropical taxa. Selected Q'eqchi' antidiabetic plants with high SIV were collected in bulk and tested in the advanced glycation endproduct (AGE) inhibition assay. All plant extracts showed AGE inhibition and the half maximal inhibitory concentration (IC50) ranged from 40.8 to 733⯵g/mL, while the most active species was Tynanthus guatemalensis Donn (Bignoniaceae). A linear regression showed a significant relationship between 1/ IC50 and SIV. Phytochemical analysis revealed the presence of verbascoside, as a major component and active principle of the T guatemalensis which had an IC50 =â¯5.1⯵g/mL, comparable to the positive control quercetin. CONCLUSION: The results reveal a rich botanical tradition of antidiabetic symptom treatments among the Q'eqchi'. Study of highly ranked plants revealed their activity in AGE inhibition correlated with SIV. T. guatemalensis was identified as a promising species for further evaluation and local use.
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Diabetes Mellitus Tipo 2/tratamento farmacológico , Hipoglicemiantes , Fitoterapia , Preparações de Plantas , Belize , Produtos Finais de Glicação Avançada/efeitos dos fármacos , Humanos , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , Medicina Tradicional , Preparações de Plantas/farmacologia , Preparações de Plantas/uso terapêutico , Plantas MedicinaisRESUMO
ABSTRACT Moussonia deppeana (Schltdl. & Cham.) Klotzsch ex Hanst., Gesneriaceae, known as tlachichinole, is a Mexican medicinal plant used for treatment of chronic inflammation-related diseases such as arthritis. In this paper, the main metabolite verbascoside was quantified in ethanolic extract; anti-arthritic and antioxidant activities were also evaluated in Complete Freund's Adjuvant induced arthritis in mice, with complete hematological evaluation, and oxidative stress measure in edema and ganglionic tissues on day 28. In popliteal ganglion, CD4+ lymphocytes and tumor necrosis factor alpha concentration were measured in addition to histological analysis. Ethanolic extract contained 79.2 mg of verbascoside/g extract, and this extract at 450 mg/kg generated an inhibition of 24% over paw edema development and increased body weight gain on final day. For hematological parameters, same dose decreased total leukocytes and lymphocytes, as well as decreased oxidation rate over biomolecules in edema and ganglionic tissues, and increased antioxidant enzyme activity. In ganglionic tissue, CD4+ lymphocytes and tumor necrosis factor alpha level showed no differences at any tested dose compared to complete Freund's adjuvant untreated group. Histological analysis of popliteal ganglion revealed moderate reduction of follicular hyperplasia, leukocyte infiltration and lipid inclusions at 450 mg/kg dose. Ethanolic extract of M. deppeana possesses anti-edematous activity associated to a moderate reduction in follicular hyperplasia, with immune-modulatory and antioxidant effects during experimental arthritis in mice.
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Ultraviolet radiation (UVR) is involved in both sunburn and the development of skin cancer, which has a high incidence worldwide. Strategies to reduce these effects include the use of photoprotective substances. The aim of this work was to investigate the photoprotective effect of verbascoside isolated from the methanolic extract of Buddleja cordata (BCME) in SKH-1 mice exposed to acute and chronic UV-B radiation. The mouse dorsal area was evaluated macroscopically and microscopically for diagnosis; verbascoside penetration into mouse skin was investigated in vivo by the tape stripping method. After acute UV-B exposure, 100 percent of irradiated mice that had been protected with verbascoside showed no signs of sunburn or of inflammatory processes. After chronic exposure, 100 percent of unprotected mice showed skin carcinomas; in contrast, in mice topically treated with either BCME or verbascoside, the presence of lesions was decreased by 90 percent. These results prove that verbascoside penetrates through the skin of mice and suggest that verbascoside and BCME may potentially prevent photodamage on mices skin after acute and chronic UVR exposure.
La radiación ultravioleta (RUV) provoca quemaduras solares y el desarrollo de cáncer de piel. El objetivo de este trabajo fue investigar el efecto fotoprotector del verbascósido obtenido del extracto metanólico de Buddleja cordata (EMBC) en ratones SKH-1 expuestos a RUV-B de manera aguda y crónica. El diagnóstico histológico se llevó a cabo en la piel de la zona dorsal de los ratones. La penetración del verbascósido fue cuantificada mediante la técnica de la cinta adhesiva. En el experimento agudo, el 100 por ciento de los ratones protegidos con verbascósido no evidenciaron signos de quemadura ni procesos inflamatorios. En el experimento crónico los ratones sin protección e irradiados presentaron carcinomas cutáneos. En contraste en los ratones protegidos con EMBC o verbascósido las lesiones disminuyeron un 90 por ciento en ambos grupos. El verbascósido penetró en la piel del ratón. Los resultados sugieren que el EMBC y el verbascósido previenen el fotodaño en la piel de ratones expuestos de forma aguda o crónica a la RUV.
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Animais , Camundongos , Buddleja/química , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Pele , Pele/efeitos da radiação , Eritema/prevenção & controle , Glucosídeos/farmacologia , Camundongos Pelados , Pele/patologia , Queimadura Solar/prevenção & controle , Raios Ultravioleta/efeitos adversosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Moussonia deppeana, known as Tlachichinole, is a Mexican medicinal plant used for treatment of inflammatory diseases, influenza, diarrhea, gastrointestinal disorders and arthritis. AIM OF THE STUDY: In this paper the antioxidant and anti-inflammatory activities as well as the acute and sub-acute toxicological effects were evaluated for the ethanolic extract from aerial parts of M. deppeana, also its phytochemical analysis is described. MATERIALS AND METHODS: Phytochemical analysis and compound isolation were performed with thin layer chromatography. The chemical identification of the main compound was performed by (1)H NMR (COSY, NOESY, HSQC and HMBC) spectra. In vitro antioxidant capacity and total phenolic content for the ethanolic extract and its primary fractions was determined by DPPH and Folin-Ciocalteu reagent. Acute and subacute toxicity tests were evaluated on Balb/C mice. Finally acute anti-inflammatory evaluation was tested for a local (TPA) and systemic (carrageenan) murine model. RESULTS: The main compound isolated from the ethanolic extract of M. deppeana was Verbascoside, which was isolated from F3 and was identified by (1)H NMR and COSY data. Furthermore oleanolic and ursolic acids were isolated from primary fractions F1 and F2. Ethanolic extract showed IC50 = 6.71mg/mL for DPPH test and 664.12µg QE/mL for the total phenolic content. The LD50 value was >2g/kg by i.g. route in male and female mice. Sub-acute administration (28 days) of the ethanolic extract (1g/kg) did not cause lethality or alter any hematological and biochemical parameters, in addition, histological analysis of the major organs exhibited no structural changes. Anti-inflammatory activity of the ethanolic extract showed an ED50 = 1.5mg/ear and 450mg/kg for TPA and carrageenan test, respectively. Primary fractions generated moderate local and systemic anti-inflammatory activity. CONCLUSION: The ethanolic extract from the aerial parts of M. deppeana did not cause any lethality or adverse effect in either of the acute and sub-acute toxicity tests. This exhibited an important local and systemic anti-inflammatory activity and also moderate antioxidant capacity. Moreover, the primary fraction F2 was more active for the TPA model while the primary fraction F3 was most active in the carrageenan model in vivo. The main compound isolated from F3 was verbascoside; on the other hand also ursolic and oleanolic acids were isolated from F1 and F2.
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Anti-Inflamatórios , Antioxidantes , Magnoliopsida , Extratos Vegetais , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Anti-Inflamatórios/toxicidade , Antioxidantes/química , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Antioxidantes/toxicidade , Compostos de Bifenilo/química , Carragenina , Edema/induzido quimicamente , Edema/tratamento farmacológico , Feminino , Glucosídeos/análise , Masculino , Medicina Tradicional , México , Camundongos Endogâmicos BALB C , Ácido Oleanólico/análise , Fenóis/análise , Ésteres de Forbol , Compostos Fitoquímicos/análise , Fitoterapia , Picratos/química , Componentes Aéreos da Planta , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Extratos Vegetais/toxicidade , Triterpenos/análise , Ácido UrsólicoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Tecoma stans is traditionally used by several ethnical groups in Mexico and Central America to treat diabetes. This species is mentioned in the majority of the ethnopharmacological studies compiled in Mexico focused in medicinal plants used as anti-diabetic treatment. AIM OF THE STUDY: Recently, this plant was found to display a high level of pancreatic lipase inhibitory activity, in addition to the several action mechanisms already described. Here we show the phytochemical and in vitro pharmacological characterization of some of the compounds responsible for the antilipase activity. MATERIALS AND METHODS: Starting with a hydroalcoholic extract, fractions were obtained by liquid-liquid separation and successive processes of column chromatography purifications. Lipase inhibitory activity was measured employing a spectrophotometric analysis. For structural elucidation (1)H and (13)C NMR experiments were used. HPLC was used to quantify and confirm the identity of the bioactive compounds. RESULTS: Bio-guided chemical purification of the hydroalcoholic extract produced an organic fraction (ethyl acetate, TsEA), flavone fractions (TsC1F13), (TsC1F15), (TsC1F16) and isolated compounds (chrysoeriol, apigenin, luteolin, and verbascoside) with the capability to inhibit the activity of pancreatic lipase. The most active fraction (TsC2F6B) was constituted by a mixture of Chrysoeriol (5,7-dihydroxy-2-[4-hydroxy-3-methoxyphenyl]chromen-4-one, 96% ) and Apigenin (4%). This flavone mixture displayed a percentage of inhibition of 85% when it was eavaluated at 0.25mg/mL. Luteolin and chrysoeriol produced a noncompetitive and mixed inhibition with values of IC50=63 and 158µM respectively. The content of chrysoeriol was also quantified in the hydroalcoholic extract (TsHAE) and organic fraction (TsEA) as 1% and 7% respectively. All of this confirms that high proportion of both flavones produce an increase of the biological activity due to they show the highest inhibition of lipase enzyme in a concentration dependant way. CONCLUSIONS: These results evidence that the medicinal use of T. stans could be in part because of its lipase inhibitory activity allowing to adapt the administration of this plant before meals. Also this data could help to develop a novel phytopharmaceutical drug (standardized in luteolin, chrysoeriol, and apigenin) auxiliary for the Type 2 Diabetes mellitus.
Assuntos
Bignoniaceae/química , Flavonas/farmacologia , Lipase/antagonistas & inibidores , Polifenóis/farmacologia , Flavonas/química , Concentração Inibidora 50 , Estrutura Molecular , Polifenóis/químicaRESUMO
Castilleja tenuiflora has been used for the treatment of several Central Nervous System (CNS) diseases. Herein we report the antidepressant activity of the methanol extract from the leaves of this medicinal plant. The oral administration of MeOH extract (500 mg/kg) induced a significant (p < 0.05) decrement of the immobility parameter on Forced Swimming Test (FST) and an increment in the latency and duration of the hypnosis, induced by administration of sodium pentobarbital (Pbi, 40 mg/kg, i.p.). Chemical analysis of this antidepressant extract allowed the isolation of (+)-piperitol-4-O-xylopyranosyl-(1â6)-O-glucopyranoside. This new furofuran lignan diglycoside was named tenuifloroside (1) and its complete chemical structure elucidation on the basis of 1D and 2D NMR spectra analysis of the natural compound 1 and its peracetylated derivative 1a is described. This compound was found together with two flavones-apigenin and luteolin 5-methyl ether-a phenylethanoid-verbascoside-and three iridoids-geniposide, caryoptoside and aucubin. All these compounds were purified by successive normal and reverse phase column chromatography. Tenuifloroside, caryoptoside and luteolin 5-methyl ether were isolated from Castilleja genus for the first time. These findings demonstrate that C. tenuiflora methanol extract has beneficial effect on depressive behaviors, and the knowledge of its chemical constitution allows us to propose a new standardized treatment for future investigations of this species in depressive illness.
Assuntos
Antidepressivos , Glicosídeos , Lignanas , Aprendizagem em Labirinto/efeitos dos fármacos , Orobanchaceae/química , Extratos Vegetais/química , Animais , Antidepressivos/química , Antidepressivos/isolamento & purificação , Antidepressivos/farmacologia , Glicosídeos/química , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Lignanas/química , Lignanas/isolamento & purificação , Lignanas/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos ICRRESUMO
A new iridoid glucoside has been isolated from the Chilean native Alonsoa meridionalis (L.f.) Kuntze. Its structure has been assigned as 6'-O-ß-d-glucopyranosyl-8-O-acetylharpagide (1) by using spectroscopic methods. Harpagoside (2), laterioside (3) and verbascoside (4) were also identified.
Assuntos
Glicosídeos Iridoides/isolamento & purificação , Scrophulariaceae/química , Chile , Glicosídeos , Glicosídeos Iridoides/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , PiranosRESUMO
The estrous cycle disruptor effect of an ethanolic extract (EMATst) from Buddleja globosa leaves and standardized in its main component (verbascoside) was determined in rats after the subcutaneous administration of EMATst. Binding of EMATst and verbascoside to the estrogen receptor (ER) of EMATst and verbacoside was also measuredestablished. EMATst produced a significant alteration inof the estrous cycle only at the highest dose (10-5 M), which could be attributed to an antiestrogenic effect. The Bbinding of EMATst and verbascoside to the ER was competitive and occurred in concentrations 1000 times greater than that of 17beta-estradiol.
El efecto disruptor del ciclo estral de un extracto etanólico (EMATst) obtenido a partir de las hojas de Buddleja globosa y estandarizado en su componente mayoritario (verbascósido) fue determinado en ratas después de la administración subcutánea de EMATst. Se estableció además la unión al receptor estrogénico (RE) tanto de EMATst como de verbascósido. EMATst sólo a la dosis más alta (10-5M) produjo una alteración significativa del ciclo estral, lo que podría atribuirse a un efecto antiestrogénico. La unión al RE de EMATst y verbascósido se produjo a concentraciones 1000 veces mayor que el 17beta-estradiol y de forma competitiva.
Assuntos
Animais , Ratos , Buddleja/química , Ciclo Estral , Extratos Vegetais/farmacologia , Fenóis/farmacologia , Glucosídeos/farmacologia , Etanol , Folhas de Planta/química , Ratos Sprague-Dawley , Receptores de EstrogênioRESUMO
Gypsophila pilulifera, Boiss & Heldr, Caryophyllaceae, is a perennial medicinal herb that grows in the southwestern region of Turkey. Except for only one report on the isolation of cytotoxic saponins from the underground parts of G. pilulifera, there are no published thorough phytochemical or bioactivity studies on this species. In the present study, the free-radical scavenging activity of extracts and fractions of the stems of G. pilulifera was evaluated, using a slightly modified and more precise version of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, reported here for the first time. The DPPH assay-guided HPLC-PDA-purification of the active solid-phase extraction fraction (50% methanol in water) of the methanolic extract exhibited verbascoside as the main free-radical scavenger present in this species. The structure of this active compound was resolved by spectroscopy, and the free-radical scavenging potential of verbascoside was determined.
RESUMO
Jacaranda cuspidifolia Mart., conhecida popularmente como "caroba", "jacarandá" ou "bolacheira", é utilizada medicinalmente para o tratamento da sífilis e da gonorréia. A atividade antimicobacteriana dessa espécie foi avaliada em ensaios in vitro com os extratos metanólicos das cascas e folhas, segundo o Método Analítico Alamar Blue (MABA). Os valores de concentração inibitória mínima para os extratos metanólicos das cascas e das folhas de J. cuspidifolia foram iguais a CIM = 250 μg mL-1 para ambos os extratos. A análise fitoquímica, por Cromatografia em Camada Delgada de gel de sílica, dos extratos metanólicos das cascas e folhas revelou a presença de taninos, flavonóides, terpenos, cumarinas e esteróides. A análise dos perfis dos extratos metanólicos por Cromatografia Líquida de Alta Eficiência de Fase Reversa registrou a presença de compostos fenólicos derivados do verbascosídeo sugerindo a provável responsabilidade pela ação antimicobacteriana.
Jacaranda cuspidifolia Mart., popularly known as "caroba", "jacaranda" or "bolacheira", is used as medicine for the treatment of syphilis and gonorrhea. The antimycobacterial activity of this species was assessed by means of in vitro assays with methanol extracts of barks and leaves according to the Microplate Alamar Blue Assay (MABA). The minimal inhibitory concentration values for methanol extracts of barks and leaves from J. cuspidifolia were MIC = 250 μg mL-1 for both extracts. Phytochemical analysis, by Thin Layer Chromatography on silica gel, of methanol extracts of barks and leaves revealed the presence of tannins, flavonoids, terpenes, cumarins and steroids. Analysis of the profiles of methanol extracts by High Performance Liquid Chromatography - Reversed Phase recorded the presence of phenolic compounds derivatives of verbascoside, suggesting their probable responsibility for the antimycobacterial action.