Effects of Qualea grandiflora Extract on the Expression of MMP-14 and HIF-1α in Cultured Fibroblasts and Preosteoblasts

Abstract Background: Qualea grandiflora (QG) (Vochysiaceae), also known as "pau-ferro", "pau-terra" or "pau-de-tucano", is a very common deciduous tree in the Brazilian Cerrado used in traditional medicine to treat inflammations, ulcers, diarrhea, and infections. There are reports in the scientific literature that demonstrate the medicinal effects of the bark and leaf of the QG. However, studies involving this plant are rather imited. Aim of the study: To perform the phytochemical analysis of the QG hydroalcoholic extract (HAE) of leaves, and to investigate it effects on fibroblast and preosteoblasts. Methods: Phytochemical analysis was done by HPLC-DAD. Murine NIH/3T3 fibroblasts and MC3T3-E1 preosteoblasts cell lines (ATCC) were used for the experiments. Cell viability was assessed by the MTT colorimetric assay and the expression of MMP-14 and HIF-1α by immunofluorescence. Results and conclusion: The following compounds were identified by HPLC-DAD, such as quinic acid, ethyl galate, ellagic acid derivatives as O-methylellagic acid O-galloyl, O-methylellagic acid O-deoxyhexoside, galloyl derivatives, flavonol glycoside as kaempferol-O-deoxyhexoside, quercetin-O-deoxyhexoside, myricetin-O-deoxyhexoside and the pentacyclic triterpene arjunglucoside. Cell viability results demonstrated no cytotoxic effects in the studied concentrations. We found in QG HAE some compounds with therapeutic properties that can increase the expression of MMP-14 and HIF-1α, in fibroblasts and preosteoblasts. These data suggest that QG HAE has an action on these two molecules widely involved in physiological conditions, such as collagen remodeling, bone development and growth and pathological processes as HIF signaling in cancer metastasis.

Disentangling the Environmental Factors That Shape Genetic and Phenotypic Leaf Trait Variation in the Tree Qualea grandiflora Across the Brazilian Savanna.

Identifying the environmental factors that shape intraspecific genetic and phenotypic diversity of species can provide insights into the processes that generate and maintain divergence in highly diverse biomes such as the savannas of the Neotropics. Here, we sampled Qualea grandiflora, the most widely distributed tree species in the Cerrado, a large Neotropical savanna. We analyzed genetic variation with microsatellite markers in 23 populations (418 individuals) and phenotypic variation of 10 metamer traits (internode, petiole and corresponding leaf lamina) in 36 populations (744 individuals). To evaluate the role of geography, soil, climate, and wind speed in shaping the divergence of genetic and phenotypic traits among populations, we used Generalized Dissimilarity Modelling. We also used multiple regressions to further investigate the contributions of those environmental factors on leaf trait diversity. We found high genetic diversity, which was geographically structured. Geographic distance was the main factor shaping genetic divergence in Qualea grandiflora, reflecting isolation by distance. Genetic structure was more related to past climatic changes than to the current climate. We also found high metamer trait variation, which seemed largely influenced by precipitation, soil bulk density and wind speed during the period of metamer development. The high degree of metamer trait variation seems to be due to both, phenotypic plasticity and local adaptation to different environmental conditions, and may explain the success of the species in occupying all the Cerrado biome.

Proteome Dataset of Qualea grandiflora Mart. (Vochysiaceae) by LC-MS/MS Label-Free Identification in Response to Aluminum.

This dataset brief is about the descriptive proteome of Qualea grandiflora plants by label free mass spectrometry (LC-MS/MS). Q. grandiflora is a plant that accumulates aluminum (Al) in high quantities and requires it for growth and development. Although quite relevant for the understanding of Al effects on plants, the proteome of Q. grandiflora has not been studied yet. Therefore, the current proteome analysis identifies a total of 2010 proteins. Furthermore, the identified Q. grandiflora root proteins are associated with several crucial molecular functions, biological processes, and cellular sites. Hence, the proteome analysis of Q. grandiflora will contribute to unravel how plants evolved to cope with high levels of Al in soils. All data can be accessed at the Centre for Computational Mass Spectrometry - MassIVE MSV000082284 - https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=adb9647282a5421a9cffe3124c060f46.

Brazilian Cerrado Qualea grandiflora Mart. Leaves Exhibit Antiplasmodial and Trypanocidal Activities In vitro.

BACKGROUND: The rapid spread of drug-resistant strains of protozoan parasites required the urgent need for new effective drugs. Natural products offer a variety of chemical structures, which make them a valuable source of lead compounds for the development of such new drugs. Cerrado is the second largest biome in Brazil and has the richest flora of all the world savannahs. We selected Qualea grandiflora, a plant species known for its proprieties in folk medicine and its antibacterial activity. OBJECTIVE: However, its antiprotozoal activity was not yet explored. MATERIALS AND METHODS: We investigated the activities of fractions from the ethyl acetate extract of Q. grandiflora leaves against human life forms of Plasmodium falciparum, Trypanosoma cruzi, and Trypanosoma brucei gambiense, and for its cytotoxicity upon the rat L6-myoblast cell line. Ten fractions were produced by ethyl acetate:hexane chromatography. RESULTS AND CONCLUSION: The fractions showed no cytotoxicity against L-6 cells (IC50 > 100 µg/mL) and no hemolysis propriety. Three fractions had a moderate activity against P. falciparum, anyone was active against T. cruzi but four fractions demonstrated a high activity against bloodstream forms of T. brucei gambiense (8.0< IC50 <15 µg/mL). Identification and characterization of the active compounds are currently under investigation. SUMMARY: Qualea grandiflora is an endemic tree of the Brazilian Cerrado, which presents medicinal propertiesTen fractions of the ethyl acetate extract of Q. grandiflora leaves were assessed against Plasmodium falciparum, Trypanosoma Cruzi, and Trypanosoma brucei gambienseNo fraction showed relevant cytotoxicity and hemolysis activityAll the fractions presented antiplasmodial and trypanocidal activitiesThree fractions with moderate antiplasmodial activity (49< IC50 <56 µg/mL)Four fractions with high activity against bloodstream forms of T. brucei gambiense (8.0< IC50 <15 µg/mL). Abbreviations used: CQ: Chloroquine, DMSO: Dimethyl sulfoxide, HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, HMI: Modified Iscove's medium, IC50: Concentration inhibiting 50% of parasite growth, IC90: Concentration inhibiting 90% of parasite growth, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, RPMI: Roswell Park Memorial Institute, SD: Standard deviation, SI: Ratio of cytotoxicity to biological activity - TC50/IC50, TC50: Concentration causing 50% of cell growth inhibition, TC90: Concentration causing 90% of cell growth inhibition, TLC: Thin-layer chromatography.

Evaluation of Qualea grandiflora Mart effect on HIF-1alpha and MMP-14 modulation in cell lines of pre-osteoblasts and fibroblasts / Avaliação do efeito da Qualea grandiflora Mart na modulação de HIF-1alpha e MMP-14 em linhagens celulares de fibroblastos e pré-osteoblastos

A espécie vegetal Qualea grandiflora (QG), popularmente conhecida como "pauferro", "pau-terra-da-folha-grande", "pau-terra" ou "pau-de-tucano", muito comum no Cerrado brasileiro, é bem conhecida devido às suas variadas propriedades terapêuticas. Suas indicações incluem ações preventivas no aparecimento de lesões de mucosa gástrica, efeitos analgésicos, antibacterianos, anti-inflamatórios e antifúngicos. Assim, os componentes da QG poderiam ter alguma ação sobre moléculas amplamente envolvidas em processos angiogênicos e de desenvolvimento/reparo, como a Metaloproteinase de matriz 14 (MMP-14) e o Fator Induzido por hipóxia 1α (HIF-1alfa). Dessa maneira, o objetivo deste estudo foi investigar os efeitos do extrato hidroalcoólico das folhas de QG na viabilidade celular e expressão de MMP-14 e HIF-1alpha em culturas de fibroblastos da linhagem NIH/3T3 e pré-osteoblastos da linhagem MC3T3-E1. Para o teste de viabilidade celular e expressão das moléculas, concentrações de 0.1; 1.0 e 10 µg/mL do extrato hidroalcoólico das folhas de QG foram administrados por períodos de 24, 48, 72 e 96h. Após cada período, a viabilidade celular foi avaliada pelo método de redução de MTT e a análise da expressão das moléculas foi feita por meio da técnica de imunofluorescência. Os resultados mostram que o extrato de QG não promove redução da viabilidade celular de fibroblastos e pré-osteoblastos em concentrações até 10 µg/mL, nos períodos iniciais (24 e 48h). Porém, uma redução significativa da viabilidade pode ser verificada nos períodos de 72h e 96h para os fibroblastos e 96h para os pré-osteoblastos, expostos a mais alta concentração do extrato (10 µg/mL). O ensaio de imunofluorescência indica que o extrato, nas concentrações de 0.1; 1.0 e 10 µg/mL foi capaz de aumentar a expressão de MMP-14 e HIF-1alpha, em ambos os tipos celulares. Em conclusão, nossos resultados indicam que o extrato de QG exerce um efeito capaz de aumentar a expressão das duas moléculas em estudo (MMP-14 e HIF-1alpha), tanto para os fibroblastos da linhagem NIH/3T3 como para os pré- osteoblastos da linhagem MC3T3-E1. Assim, os compostos de QG podem apresentar potencial para serem utilizados como agentes terapêuticos moduladores da angiogênese, por meio do aumento da expressão de MMP-14 e HIF-1alpha.(AU)

The vegetable specie Qualea grandiflora (QG), popularly known as "pau-ferro", "pauterra-da-folha-grande", "pau-terra" or "pau-de-tucano", very common in the Brazilian Cerrado, is well known due to its varied therapeutic properties. Its indications include preventive actions in the appearance of lesions of gastric mucosa, analgesic, antibacterial, anti-inflammatory and antifungal effects. Thus, QG components could have some action on molecules widely involved in angiogenic and developmental / repair processes, such as Matrix metalloproteinase 14 (MMP-14) and HypoxiaInducible Factor-1α (HIF-1alpha). Thus, the objective of our study was to investigate the effects of QG hydroalcoholic extract on cell viability and expression of MMP-14 and HIF-1alpha in NIH/3T3 fibroblasts and MC3T3-E1 pre-osteoblasts cell lines. For the cell viability assay and expression of the molecules, concentrations of 0.1; 1.0 and 10 µg / mL of the hydroalcoholic extract of leaves of QG, were administered for periods of 24, 48, 72 and 96h. After each period, the cell viability was evaluated by MTT assay and the expression of the molecules was analyzed using the immunofluorescence technique. The results show that the QG extract does not promote reduction of the cellular viability of fibroblasts and pre-osteoblasts in concentrations up to 10 µg/mL in the initial periods (24 and 48h). However, a significant reduction in viability can be observed in 72h and 96h for fibroblasts and 96h for pre-osteoblasts exposed to the highest extract concentration (10 µg/mL). The immunofluorescence assay indicates that the extract, at concentrations of 0.1; 1.0 and 10 µg/mL was able to increase the expression of MMP-14 and HIF-1alpha in both cell types. In conclusion, our results indicate that the QG extract exerts an effect capable of increasing the expression of the two molecules under study (MMP-14 and HIF-1alpha) both for the NIH/3T3 fibroblasts as well as for the MC3T3-E1 pre-osteoblasts cells. Thus, the QG compounds could have potential to be used as angiogenesis modulating therapeutic agents, by increasing the expression of MMP-14 and HIF-1alpha.(AU)

Concentração inibitória mínima e concentração bactericida mínima de extratos hidroalcoólicos das folhas de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre Streptococcus mutans e Lactobacillus casei / Minimum inhibitory concentration and minimum bactericidal concentration of hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans and Lactobacillus casei

A cárie dentária é uma doença bucal de alta prevalência e impactante em países em desenvolvimento. É causada pela presença de biofilme dentário rico em bactérias acidogênicas e acidúricas, como Streptococcus mutans e Lactobacillus casei. Neste sentido, a fitoterapia tem sido aplicada na odontologia devido ao seu conhecido efeito antimicrobiano, tendo potencial para prevenir doenças como a cárie dentária. Portanto, o presente estudo tem como objetivo testar o potencial antimicrobiano de extratos bruto e etanólico das folhas de Myracrodruon urundeuva (M. urundeuva.) e Qualea grandiflora (Q. grandiflora) sobre S.mutans e L. casei. Para tal, determinaram-se a Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM). A CIM foi definida como a menor concentração do agente antimicrobiano capaz de inibir 100% o crescimento microbiano (absorbância) em relação aos controles negativos. Para a CBM, alíquotas foram removidas dos poços que não apresentaram nenhuma absorbância (viabilidade, concentrações CIM) e semeadas em placas de ágar BHI, incubadas por 24 h a 37°C em estufa de CO2 5%. A CBM foi determinada considerando a menor concentração dos extratos capaz de impedir o crescimento bacteriano visível. Cepas de S. mutans (ATCC 21175) e L. casei (ATTC 334) foram ativadas em BHI e caldo Rogosa, respectivamente. A CIM foi determinada pela técnica de diluição em microplacas de 96 poços (100 l de extrato + 80 l BHI/Rogosa + 20 l da bactéria diluída em BHI/Rogosa equivalente a 5x105 UFC/mL), as quais foram incubadas por 24 h (S. mutans) e 48 h (L. casei) a 37°C em estufa de CO2 5%. Os extratos de M. urundeuva e Q. grandiflora inicialmente foram diluídos em BHI/Rogosa variando as concentrações entre 2 mg/ml a 0,00012207 mg/ml e os mesmos extratos diluídos em álcool foram avaliados nas concentrações entre 20 mg/ml a 0,00244 mg/ml para S. mutans e L. casei. Não foi possível determinar a CIM e a CBM para os extratos diluídos no BHI/Rogosa. Foram utilizados como controle positivo a clorexidina e como controles negativos BHI/Rogosa com e sem álcool a 5%. As CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e clorexidina contra S. mutans foram 2,5 mg/ml (2,5 mg/ml), 5,0 mg/ml (--) e 0,00468 mg/ml (0,00937 mg/ml), respectivamente. Em relação ao L. casei, as CIMs (CBMs) da M. urundeuva e Q. grandiflora, diluídas em álcool, e da clorexidina foram 0,156 mg/ml (0,312 mg/ml), 0,156 0,625 mg/ml (0,312 0,625mg/ml) e 0,00468 mg/ml (0,3 mg/ml), respectivamente. Como conclusão, nosso estudo mostrou que L. casei (ATTC 334) é mais susceptível aos extratos que S. mutans (ATCC 21175) e o extrato M. urundeuva apresenta melhor efeito antimicrobiano que a Q. grandiflora em S. mutans (ATCC 21175), porém os dois extratos apresentam efeito similar sobre L. casei (ATTC 334) e ambos foram inferiores à CHX.(AU)

Dental caries is an oral disease of high prevalence and impact in developing countries. It is caused by the presence of a dental biofilm rich in acidogenic and aciduric bacteria, such as Streptococcus mutans and Lactobacillus casei. Accordingly, phytotherapy has been applied in dentistry due to its known antimicrobial effect, having potential to prevent diseases such as dental caries. Therefore, the present study aims to test the antimicrobial potential of crude and ethanolic extracts of Myracrodruon urundeuva (M. urundeuva) and Qualea grandiflora (Q. grandiflora) leaves on S. mutans and L. casei. For this, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. MIC was defined as the lowest concentration of the antimicrobial agent capable of inhibiting 100% the microbial growth in comparison to the negative controls. For MBC, aliquots were removed from the wells that did not show any absorbance (viability, concentrations than MIC) and seeded on BHI agar plates, incubated for 24 h at 37°C and 5% CO2. The MBC was determined considering the lowest concentration of extracts capable of preventing visible bacterial growth. Strains of S. mutans (ATCC 21175) and L. casei (ATTC 334) were activated in BHI and Rogosa broth. MIC was determined by the dilution technique in 96-wells microplates (100 l of extract + 80 l BHI/ Rogosa + 20 l of bacterium diluted in BHI/Rogosa equivalent to 5x105 CFU/ml), which were incubated for 24 h (S. mutans) and 48 h (L. casei) at 37°C and 5% CO2. The extracts were firstly diluted in BHI/Rogosa varying the concentrations between 2 mg/mL and 0.00012207 mg/ml; the same extracts were also diluted in alcohol at concentrations ranging from 20 mg/ml to 0.00244 mg/ml and both tested against S. mutans and L. casei. It was not possible to determine the MIC and MBC for the extracts diluted in BHI/Rogosa. Chlorhexidine was used as positive control, while BHI/Rogosa with or without 5% alcohol were used as negative controls. The MICs (MBCs) of M. urundeuva and Q. grandiflora diluted in alcohol, and clorexidine against S. mutans were 2.5 mg/ml (2.5 mg/ml), 5.0 mg/ml (--) and 0.00468 mg/ml (0.00937 mg/ml), respectively. In respect to L. casei, the MICs (MBCs) of M. urundeuva and Q. grandiflora, diluted in alcohol, and chlorhexidine were 0.156 mg/ml (0.312 mg/ml), 0.156 0.625 mg/ml (0.312 0.625 mg/ml) and 0.00468 mg/ml (0.3 mg/ml), respectively. In conclusion, our study showed that L. casei (ATTC 334) is more susceptible than S. mutans (ATCC 21175) to the extracts and the extract of M. urundeuva has a better antimicrobial effect than Q. grandiflora against S. mutans (ATCC 21175), but both extracts have similar effect on L. casei (ATTC 334) and they were inferior to CHX.(AU)

Reversed-phase-liquid chromatography method for separation and quantification of gallic acid from hydroalcoholic extracts of Qualea grandiflora and Qualea parviflora.

BACKGROUND: Qualea parviflora and Qualea grandiflora (Vochysiaceae), commonly known in Brazil as "pau-terra" and "pau-terrinha," respectively, have been widely used in the treatment of ulcer and gastritis. These therapeutic effects are attributed to various compounds present in the plants, including phenolic compounds such as gallic acid, due to their important antioxidant activity. OBJECTIVE: The aim of the present study was to validate a high performance liquid chromatography with diode array detection (HPLC-DAD) method for the quantitative determination of gallic acid in the stem bark of Q. parviflora and Q. grandiflora hydroalcoholic extracts. MATERIALS AND METHODS: The chromatography analysis was successfully achieved on a Dionex column, Acclaim(®) 120 (250 mm × 4.60 mm, 5 µm) with a gradient elution of water and methanol at a flow rate of 0.8 mL/min and ultraviolet detection at 280 nm. RESULTS: The validation data, including linearity, precision, specificity, accuracy and robustness of this method demonstrated good reliability and sensitivity. CONCLUSION: The method is able to quantify gallic acid in the stem bark of both species. What is more, the chromatographic peaks showed good resolution and there are also the advantages of easy sample preparation and a short time between each injection.

Atividade antibacteriana de plantas úteis e constituintes químicos da raiz de Copernicia prunifera / Antibacterial activity of useful plants and chemical constituents of the roots of Copernicia prunifera

Extratos etanólicos de Qualea grandiflora e Copernicia prunifera e extrato hexânico de Dipteryx lacunifera foram avaliados quanto a atividade antibacteriana, utilizando ensaios de difusão a partir de orifício e concentração inibitória mínima (CIM), frente a cepas Gram positivas e Gram-negativas, incluindo espécies multidroga resistentes. O extrato de Q. grandiflora apresentou atividade moderada para as cepas de Staphylococcus epidermidis (CIM = 500 µg/mL) e atividade fraca sobre as demais bactérias Gram-positivas testadas e inativo sobre bactérias Gram-negativas. Os resultados obtidos com S. epidermidis apesar de moderados são importantes, uma vez que este microorganismo é o principal causador de bacteremias e sepse associada com dispositivos médicos implantados. gama-Tocoferol e a mistura de sitosterol e estigmasterol foram isolados do extrato etanólico de raiz de C. prunifera e as estruturas destes compostos foram identificadas com base na análise dos dados espectrais de RMN e comparação com a literatura.

Ethanol extracts of Qualea grandiflora and Copernicia prunifera and hexane extract of Dipteryx lacunifera were evaluated by diffusion in agar and minimum inhibitory concentration (MIC) against Gram-positive and Gram-negative bacteria including multiresistant drug strains. The extract of Q. grandiflora presented moderate activity for Staphylococcus epidermidis (MIC = 500 µg/mL) and weak activity against other Gram-positive strains and inactive for Gram-negative species. The results obtained for S. epidermidis despite being moderate are important because this pathogen is often recovered from bacteremia and sepsis from infections of implanted devices. gamma-Tocopherol and the mixture of sitosterol and stigmasterol were isolated from the ethanol extract of the roots of C. prunifera. The structures of these compounds were identified by NMR spectroscopy and comparison with literature data.

The effects of phosphate supply on growth of plants from the Brasilian Cerrado: experiments with seedlings of the annual weed, Bidens gardneri Baker (Compositeae) and the tree, Qualea grandiflora (Mart.) (Vochysiaceae).

Plants of the cerrado tree species Qualea grandiflora and the annual herb Bidens gardneri were grown from seed in controlled environment rooms at 30/20° C and 12 hour photoperiod. Seedlings were grown in pots or small tubes containing sand and provided with various amounts of mineral solutions based on the formulation of Hoagland and Arnon but with the phosphate content modified in some cases. In a long-term experiment lasting 213 days, plants supplied with full strength Hoagland's solution all died but plants of Qualea given 1/10 strength solution survived, although they grew very slowly. Low relative growth rates (0.008-0.036 d-1) were also a feature of other experiments with Qualea and calculated rates of net assimilation rate gave values of 3-7 mg CO2 dm-2 h-1. Expansion of the photosynthetic surface proceeded slowly and the cotyledons were the main site of photosynthesis for more than 40 days. The low rates of growth occurred despite significant uptake of phosphorus by young plants and in shortterm experiments growth was independent of the amount of phosphate supplied and accumulated. In contrast, the values of R found for plants of Bidens reached 0.24 d-1. Growth of young plants was dependent on the external supply of phosphorus, being reduced when this was low and also when it was very high. Growth of the photosynthetic surface was also much more rapid than for Qualea and also varied with supply of phosphorus. The results are discussed in the context of the occurrence of these species in the Cerrado.