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.

Toxicological, genotoxic and antioxidant potential of pyrostegia venusta / Potencial toxicológico, genotóxico e antioxidante de Pyrostegia venusta

Pyrostegia venusta is usually found in the secondary growth of the Atlantic forests, and in the Brazilian Savanna. Flowers and leaves of this plant are used in folk remedies for treating a wide variety of healthy conditions, this way is important evaluate its safety and antioxidant potential for this applications. For this, was made a ethanolic extract from its flowers and analyzed with toxicological,genotoxicity and antioxidant tests, the toxicological analysis was made by reproductive toxicity in rats and clatogenicity/aneugenicity in human lymphocytes. The genotoxicity was studied by micronucleus test mice bone marrow. The antimutagenic test in root cells of Allium cepa, the antioxidant assays used was DPPH, FRAP, Lipid Perxidation and REM, beyond of that the extract was analyzed in HPLC showing the profile of its compounds. The toxicological analysis showed that P. venusta has no negative significant effect on reproductive and cellular level. The micronucleus test in mouse bone marrow, the extract protected cells from cyclophosphamide, mutagenic compound, in a similar way. The A. cepa test showed that the extract reduced chromosomal disorders formations. The antioxidant activity of extract was significant, except in REM test. The phytochemical analysis showed the presence of flavonoids compounds. P. venusta extract does not present reproductive toxicity and genotoxic effects. However, the extract of this species showed antigenotoxic and antioxidant potential, possibly due to the different flavonoid compounds present in its extract.

Pyrostegia venusta é geralmente encontrada no crescimento secundário das florestas atlânticas e na savana brasileira. Flores e folhas desta planta são utilizadas em remédios populares para tratar uma grande variedade de doenças, desta forma é importante avaliar a segurança e o potencial antioxidante para estas aplicações. Para tanto, o extrato etanólico das flores foi avaliado com testes toxicológicos, genotóxicos e antioxidants. A análise toxicológica foi realizada por meio da toxicidade reprodutiva em ratos e a clatogenicidade/aneugenicidade em linfócitos humanos, a genotoxicidade foi estudada por teste de micronúcleo em medula óssea de camundongo. A antimutagenicidade em células da raiz de Allium cepa. Os ensaios antioxidantes utilizados foram DPPH, FRAP, TARBS e MRE. O extrato foi analisado em HPLC. A análise toxicológica reprodutiva mostrou que P. venusta não tem efeito negativo sobre o nível reprodutivo e cellular. No teste do micronúcleo o extrato protegeu as células da ciclofosfamida, um composto mutagênico. O teste de A. cepa mostrou que o extrato reduziu as formações dos distúrbios cromossômicos. A atividade antioxidante do extrato foi significativa, exceto no teste REM. A análise fitoquímica mostrou a presença de compostos flavonoídicos. O extrato de P. venusta não apresenta toxicidade reprodutiva e efeitos genotóxicos. No entanto, o extrato desta espécie apresentou potencial antigenotóxico e antioxidante, possivelmente devido aos diferentes compostos flavonoídicos presentes em seu extrato.

Viability of Human Gingival Fibroblast (FGH) Treated with Ethanolic "Aroeira" Extract (Myracrodruon urundeuva Allemão)

ABSTRACT The aim of this study was to evaluate the effect of ethanolic "aroeira" (Myracrodruon urundeuva) extract on the viability of human gingival fibroblast. For this, fibroblasts (2x103 cells/well) were plated in a 96-well plate and incubated for 24 h; the medium (Eagle's medium modified by Dulbecco - DMEM) supplemented with 10% fetal bovine serum was replaced by DMEM with different ethanolic extract concentration (0, 0.1, 1, 10, 100, and 1000μg / mL). The fibroblast viability was analyzed after 48,72, and 96 h by the neutral red capture test and violet crystal. The "aroeira" extract, at high concentrations (100 and 1000 µg/mL) caused decrease in both cellular viability tests (p<0.05). However, dilutions between 0.1 and 10 µg/mL did not affect the viability of the cells. It was concluded that "aroeira" extract was able to change the gingival fibroblast viability, and this effect was concentration dependent.

Growth and physiological responses of sunflower plants exposed to ultraviolet-B radiation

The effects of UV-B radiation were studied in sunflower plants (Helianthus annuus L. cv. Catissol-01) growning in greenhouse under natural photoperiod conditions. The plants received approximately 0.60Wm-2 (control) or 4.0Wm-2 (+UV-B) of UV-B radiation for 7h d-1, centered around solar noon from 15 days after sowing. Compared to the control, plants exposed to high UV-B radiation for 12 or 21 days did not show any difference in shoot dry matter, specific leaf weight or UV-B absorbing compounds. Enhanced UV-B radiation caused a significant inhibition of photosynthesis (A) only in the first sampling and this was accompained by reduction in stomatal conductance (g s) and transpiration rate. The inhibition in A can not be fully explained by reduction in g s since intercellular CO2 concentration was not affected by UV-B radiation. In both samplings, the total chlorophyll content was not affected by enhanced UV-B radiation whereas in the first sampling, the chlorophyll a and the ratio of chlorophyll a/b were reduced. Enhanced UV-B radiation increased the minimal fluorescence yield, but did not alter the ratio of variable to maximal fluorescence yield of dark adapted leaves. Overall, this study suggests that the present level of solar UV-B radiation affects sunflower plants performance even though the shoot dry biomass may not be affected.

Os efeitos da radiação UV-B foram estudados em plantas de girassol (Helianthus annuus L. cv. Catissol-01) cultivadas em casa de vegetação sob condições fotoperiódicas naturais. As plantas receberam aproximadamente 0,60W m-2 (controle) ou 4,0W m-2 (+UV-B) de radiação UV-B por 7h d-1, centralizadas ao redor do meio-dia. A irradiação com UV-B foi iniciada 15 dias após a semeadura. Plantas sob alta radiação UV-B durante 12 ou 21 dias não apresentaram diferenças em matéria seca da parte aérea, peso foliar específico ou compostos que absorvem UV-B, quando comparadas com o controle. Alta radiação UV-B reduziu a taxa de fotossíntese (A) somente na primeira coleta, sendo acompanhada por uma redução na condutância estomática (g s) e na taxa de transpiração. A inibição da fotossíntese não pode ser somente explicada pela redução na g s, visto que a concentração intercelular de CO2 não foi alterada pelo aumento na radiação UV-B. O conteúdo total de clorofila não foi afetado pelo aumento na radiação UV-B nas duas coletas. No entanto, o conteúdo de clorofila a e a razão clorofila a/b foram reduzidas na primeira coleta. Sob alta radiação UV-B, houve aumento na fluorescência mínima na primeira coleta. Porém, a razão entre a fluorescência variável e a fluorescência máxima das folhas adaptadas ao escuro não foi alterada. Estes resultados sugerem que o atual nível de radiação solar UV-B afeta o desempenho das plantas de girassol, embora a massa seca da parte aérea não seja afetada.