Effect of silencing sirtuin 3 on resveratrol-induced apoptois of human ovarian cancer SKOV3 cels / 吉林大学学报(医学版)

Objective: To investigate the effect of silencing sirtuin 3 (Sirt3) on the apoptosis of human ovarian cancer SKOV3 cells induced by resveratrol (Res), and to explore its mechanism of promoting apoptosis. Methods: The human ovarian cancer SKOC3 cells were cultured with different concentrations 0, 2. 5, 5. 0, 10.0, 20.0, 40.0 and 80.0 mg · L-1) of Res for 24 h. The survival rate of cells was measured by MTT assay. The SKOV3 cells were randomly divided into control group, Sirt3 inhibitory 3-1H-1, 2, 3-triazol-4-yl) pyridine 3-TYP group, Res group and 3-TYP+Res group. After 24 h of culture, the inhibitory rates of proliferation of the cells in various groups were detected by MTT assay; the nuclei were stained with Hoechst 33342, and the morphorgy nucleus was observed by laser confocal microscope; reactive oxygen species (ROS) probe was used to detect the intracellular ROS levels; Western blotting method was used to detect the expression levels of Sirt3, Bax, Bcl-2 and cleaved caspase-3 proteins in the cells in various groups. Results: The results of MTT assay showed that the survival rates of SKOV3 cells were significantly decreased with the increase of concentration of Res, and the median inhibitory concentration (IC50) was 42. 73 mg · L-1. Compared with control group, the inhibitory rates of proliferation of cells in Res group and 3-TYP+Res group were significantly decreased (P0.05); the protein expression levels of Sirt3 and Bcl-2 proteins in Res group were significantly decreased (P< 0.05), and the expression levels of Bax and cleaved caspase-3 proteins were significantly increased (P<0.05). Compared with Res group, the expression levels of Bax and cleaved caspase-3 proteins in 3-TYP + Res group were significantly increased (P<0.05), and the expression levels of Bcl-2 and Sirt3 proteins in 3-TYP+Res group were significantly decreased (P<0.05). Conclusion: Res can induce the apoptosis of SKOV3 cells, and the inhibition of Sirt3 expression by 3-TYP can enhance the effect of Res.

Compostos carbonílicos α, ß-insaturados e hidrazonas sintéticas como antiparasitários / α, ß-Unsaturated carbonyl compounds and Synthetic hydrazones as antiparasitics

Apesar da grande relevância médica e social, e por serem responsáveis por grande parte das mortes em países subdesenvolvidos e em desenvolvimento as doenças negligenciadas (DN), ainda, não apresentam terapêutica eficaz. Dentre as diversas DN, doenças como a doença de Chagas, a Leishmaniose visceral e a malária, se destacam no cenário nacional, por terem alta incidência e prejuízos sociais. Os fármacos disponíveis para o tratamento destas parasitoses, apresentam alta toxicidade e, em alguns casos, resistência por parte dos parasitas. Assim sendo, faz-se necessário o planejamento e desenvolvimento de novos agentes quimioterápicos mais seguros e eficazes. Dentre as diferentes estratégias de planejamento de fármacos, selecionamos o planejamento de fármacos baseado na estrutura do ligante - LBDD (Ligand-Based Drug Design) - como base para desenvolvimento deste trabalho. Nesta estratégia, utiliza-se o conhecimento de moléculas (ligantes) e de suas atividades biológicas conhecidas previamente determinadas experimentalmente, como protótipos para a busca de novas entidades químicas com atividade biológica semelhante ou melhorada. Sendo assim, o presente trabalho teve como objetivo a síntese e avaliação biológica de moléculas bioativas para o tratamento de doenças parasitárias. Baseando-se no conhecimento prévio da atividade antiparasitária de compostos carbonílicos α,ß-insaturados e hidrazonas, foram sintetizados séries de compostos destas classes químicas na busca de novos agentes quimioterápicos. Os compostos obtidos foram avaliados contra a forma epimastigota de Trypanosoma cruzi, promastigota de Leishmania donovani, amastigota de Leishmania infantum e, também, determinou-se o seu grau de citotoxicidade (CC50) frente a células de macrófago humanos diferenciado (THP-1). As 31 moléculas obtidas foram caracterizadas por técnicas de ponto de fusão, RMN 1H e RMN 13C e avaliada sua pureza por HPLC. Os compostos da classe da cinamoil-hidrazonas apresentaram-se como promissores antiparasitários, mostrando atividade frente a forma promastigota (Leishmania donovani), 4 dos 12 compostos foram ativos (IC50= 1,27 - 13,68 µM) e frente a forma amastigota (Leishmania infantum), 10 dos 12 compostos apresentaram atividade (9,09 - 63,5 µM). Mesmo apresentando citotoxicidade moderada (CC50 = 8,83 - 87,47 µM), os compostos obtiveram valores inferiores ao fármaco de referência (doxorubicina: CC50 = 0,26 µM). Diante do exposto, o planejamento de fármacos realizado por LBDD mostrou-se bem-sucedido, pois a classe de cinamoil-hidrazonas mostrou-se promissora como antiparasitários, visto sua atividade na escala de baixo micromolar e moderada citotoxicidade em células humanas. Esses resultados assinalam que a classe de compostos descrita está passível a continuar sendo investigada no intuito de aprimorar os protótipos obtidos na busca de novos agentes quimioterápicos antiparasitários e desvendar os mecanismos de ação leishmanicida

Despite to the great medical and social relevance and the amount of deaths in underdeveloped and developing countries, neglected diseases (ND) still do not have an effective therapy. Among the various ND, illnesses such as Chagas disease, visceral leishmaniasis and malaria holds a great importance in the Brazilian scenario due to high incidence and social damage. The drugs available for the treatment of these parasitosis present high toxicity and, in some cases, resistance by the pathogens. Thus, the planning and development of new, safer and more effective chemotherapeutic substances are urgent needed. Among the different drug planning strategies, we selected ligand-based drug design (LBDD) as the basis for the development of this work. In this strategy, we use the knowledge of molecules (ligands) and their known biological activities previously determined experimentally, as prototypes to search for new chemical entities with similar or improved biological activity. Therefore, the present work aimed the synthesis and biological evaluation of bioactive molecules for the treatment of parasitic diseases. Based on previous knowledge of the antiparasitic activity of α,ß-unsaturated and hydrazone carbonyl compounds, series of compounds of these chemical classes were synthesized in search of new chemotherapeutic agents. The compounds obtained were evaluated against the epimastigote form of Trypanosoma cruzi, Leishmania donovani promastigote, Leishmania infantum amastigote and their cytotoxicity (CC50) against differentiated human macrophages (THP-1). The 31 molecules obtained were characterized by melting point, 1 H NMR and 13C NMR techniques and their purity were characterized by HPLC. The cinnamoyl hydrazone class compounds showed promising antiparasitic activity, showing activity against promastigote form (L. donovani), 4 of 12 compounds were active (IC50 = 1.27 - 13.68 µM) and amastigote form (L. infantum), 10 of the 12 compounds showed activity (9.09 - 63.5 µM). Even presenting moderate cytotoxicity (CC50 = 8.83 - 87.47 µM), the compounds had values below the reference drug (doxorubicin: CC50 = 0.26 µM). Considering the results, LBDD drug planning proved to be successful and the class of cinnamoyl hydrazones were promising as antiparasitics due to its activity in low micromolar scale and moderate cytotoxicity in human cells. These results indicate that the described class of compounds can be further investigated in order to improve the prototypes obtained in the search for new antiparasitic chemotherapeutic agents and to unravel the mechanisms of action of leishmanicidal molecules

Evaluation as antioxidant agents of 1,2,4-triazole derivatives: effects of essential functional groups.

A series of 1,2,4-triazole derivative compounds substituted with groups of phenol and pyridine were synthesized in high yields and screened against several antioxidant activity parameters such as DPPH, ABTS, metal chelating, reducing power and the total antioxidant activity. The compounds showed better than expected antioxidant activity between the studied biological activity parameters. Among these, compound G(2-(5- mercapto-4H-1,2,4-triazol-3-yl)phenol) had a high total antioxidant activity potential with value of 232.12±6.89 mmol/ml. Also showed fairly good ABTS cation radical and DPPH radical scavenging activity with values of IC50 = 4.59±4.19 and IC50 = 7.12±2.32μg/mL respectively. Further, the antioxidant potential of heterocyclic compounds that 1,2,4-triazole derivatives containing different functional groups were compared with various tests performed and has been shown to increase the activity of electron donating groups. Therefore, the present study demonstrates that phenol and pyridine substituted 1,2,4-triazole compounds would be a better prospective in the development of antioxidant agent.

Espectros de gotas e idade de trifólios e sua interação com a área abaixo da curva de progresso da ferrugem da soja / Drops spectra and leaflets age and its interaction with the area under disease progress curve of soybean rust

O objetivo do presente trabalho foi avaliar o efeito de espectros de gotas (grossas, médias e finas) na velocidade de absorção de fungicidas, para trifólios de diferentes idades, através de medida indireta expressa pelo residual de controle de ferrugem asiática da soja (Phakopsora pachyrhizi). O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, em arranjo fatorial (3x3x5x4), cujos fatores foram compostos por: três espectros de gotas (grossas, médias e finas); três idades fisiológicas de trifólios na planta (1°, 3° e 5° trifólios); quatro períodos de tempo entre a aplicação de fungicidas e chuva simulada (0, 30, 60 e 120min.), mais uma testemunha sem chuva; três tratamentos com fungicida: azoxistrobina + ciproconazol (60+24g i.a. ha-1) + óleo mineral 0,6L ha-1, azoxistrobina (50g i.a. ha-1) + óleo mineral 0,6L ha-1 e ciproconazol (30g i.a. ha-1), mais uma testemunha sem aplicação. Avaliou-se densidade de gotas por centímetro quadrado, diâmetro mediano volumétrico, diâmetro mediano numérico, amplitude relativa, número de dias para o aparecimento da primeira pústula e a área abaixo da curva de progresso da doença. Verificou-se que gotas de menor DMV proporcionam maior velocidade de absorção de fungicidas. Trifólios mais novos absorvem os fungicidas mais rapidamente. A utilização da mistura de azoxistrobina + ciproconazol proporcionou menor evolução da doença com consequente menor área abaixo da curva de progresso da doença.

This research aimed to evaluate the effect of large, medium and fine droplets spectra and its interaction with the fungicide absorption rate and leaflets age through indirect measurement expressed by the residual of Asian soybean rust (Phakopsora pachyrhizi) control. The experimental design was composed by completely randomized design with four replications in a factorial (3x3x5x4), which factors were composed of: three drops spectra (large, medium and fine); three fisiological leaflets ages setting on the plant (1st, 3rd and 5th leaflets), four time periods between the fungicide application and simulated rain (0, 30, 60 and 120') and a control with no rain; three fungicide treatments with Azoxystrobin+Cyproconazole (60+24g a.i. ha-1) + 0.6L ha-1, Azoxystrobin (50g a.i. ha-1) + 0.6L ha-1 and Cyproconazole (30g a.i. ha-1), and control without spray. It were evaluated the density of droplets per square centimeter, volume median diameter, number median diameter, relative amplitude, number of days for the first pustule display and the area under disease progress curve. It was found that drops with smaller volume median diameter have higher fungicides absorption rates. Newer leaflets provide more quickly fungicides absorption. The use of Azoxystrobin + cyproconazole provided lower disease progress with consequent lower area under the disease progress curve.

Espectro de gotas e idade de trifólios na taxa de absorção e efeito residual de fungicidas em soja / Drops spectra and leaflets age on the fungicides absorption rate and residual effect in soybean

O objetivo do presente trabalho foi avaliar o efeito de espectros de gotas (grossas, médias e finas) na velocidade de absorção de fungicidas, para trifólios de diferentes idades através de medida indireta expressa pelo residual de controle de ferrugem asiática da soja (Phakopsora pachyrhizi). O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, em arranjo fatorial (3x3x5x4), cujos fatores foram compostos por: três categorias de gotas (grossas, médias e finas); três diferentes posições de avaliação na planta (1°, 3° e 5° trifólios); quatro períodos de tempo entre a aplicação de fungicidas e aplicação de chuva simulada (0, 30, 60 e 120min), mais uma testemunha sem chuva; testemunha sem aplicação, mais os fungicidas azoxistrobina + ciproconazol (60+24g i.a. ha-1) + óleo mineral 0,6L ha-1, azoxistrobina (50g i.a. ha-1) + óleo mineral 0,6L ha-1 e ciproconazol (30g i.a. ha-1). Avaliou-se a densidade de gotas por centímetro quadrado, diâmetro mediano volumétrico, diâmetro mediano numérico e amplitude relativa, além do número de dias para o aparecimento da primeira pústula. Verificou-se que gotas de menor DMV proporcionam maior velocidade de absorção de fungicidas. Trifólios mais novos absorvem os fungicidas mais rapidamente. A utilização de azoxistrobina + ciproconazol + óleo mineral proporcionou o maior número de dias até o aparecimento da primeira pústula.

This research aimed to evaluate the effect of large, medium and fine droplets spectra and its interaction with the fungicide absorption rate and leaflets age through indirect measurement expressed by the residual control of Asian soybean rust (Phakopsora pachyrhizi). The experimental design was completely randomized with four replications in a factorial scheme (3x3x5x4), which factors were composed of: three drops spectra (large, medium and fine); three different positions in the plant evaluated (1st, 3rd and 5th leaflets), four time periods between the fungicide application and simulated rain (0, 30, 60 and 120') and a control with no rain; control without fungicide application, Azoxystrobin+Cyproconazole (60+24g a.i. ha-1) + 0.6L ha-1, Azoxystrobin (50g a.i. ha-1) + 0.6L ha-1 and Cyproconazole (30g a.i. ha-1). It was evaluated the density of droplets per square centimeter, volume median diameter, number median diameter and relative amplitude, and the number of days for the first pustule display. It was found that drops with smaller volume median diameter have higher fungicides absorption rates. Newer leaflets provide more quickly fungicides absorption. The use of Azoxystrobin + Cyproconazole provided the greatest number of days until the appearance of the first pustule.

Efeito de herbicidas e fungicidas nas curvas de progresso e quantificação de danos causados pela ferrugem asiática da soja / Effect of herbicides and fungicides on progress curves and damage quantification caused by soybean rust

O fungo Phakopsora pachyrhizi Sydow & Sydow representa grande ameaça à sojicultura nas principais regiões produtoras do mundo, onde significativas perdas foram relatadas. Na quantificação de danos causados, muitas são as variáveis a serem estudadas, como área abaixo da curva de progresso da doença (AACPD), absorção de luz da área foliar sadia (HAA) e duração da área foliar sadia (HAD), além das curvas de progresso da doença. Neste trabalho, objetivou-se verificar a influência de diferentes herbicidas e fungicidas no progresso da doença, bem como suas implicações nas variáveis referentes à área foliar sadia e à produtividade da cultura. Os tratamentos testados em duas cultivares ('MG/BR46 Conquista' e 'BRS Valiosa RR') foram: 1. Testemunha; 2. Testemunha com herbicidas; 3. Herbicidas e pyraclostrobin (V4) + pyraclostrobin + epoxiconazole (R2 e R5.1); 4. Herbicidas e pyraclostrobin (V4) + epoxiconazole (R2 e R5.1); e 5. Herbicidas (V4) + pyraclostrobin + epoxiconazole (R2 e R5.1). Os herbicidas utilizados em 'MG/BR-46 Conquista' foram sethoxydim, bentazon e chlorimuron-ethyl, sendo utilizado glyphosate na 'BRS Valiosa RR'. O modelo logístico foi o que melhor se adequou à severidade média das avaliações e à curva de progresso da doença. Para o terço inferior, posição do dossel mais propícia à doença, o modelo logístico com taxa variável foi o que mais se ajustou aos dados. A pulverização de pyraclostrobin + epoxiconazole reduziu a taxa de desenvolvimento de P. pachyrhizi em relação à epoxiconazole. A utilização da mistura influenciou as variáveis AACPD, HAA e HAD, sendo estas consideradas apropriadas para a quantificação dos danos provocados pela ferrugem asiática. Na 'MG/BR-46 Conquista', a ação dos herbicidas afetaram temporariamente a área foliar, atingindo indiretamente as variáveis HAA e HAD.

Phakopsora pachyrhizi Sydow & Sydow is the worst threat for soybean crop in the most important growing regions around the world, where great losses were observed. Many variables are studied in the quantification of damage such as area under the disease progress curve (AUDPC), healthy leaf area absorption (HAA), healthy leaf area duration (HAD), in addition to disease progress curves. The present research aimed to evaluate the effect of several herbicides and fungicides on the progress of the disease and its implications for variables related to healthy leaf area and yield. The treatments were evaluated in two cultivars (MG/BR-46 Conquista e BRS Valiosa RR): 1. Control; 2. Control with herbicides; 3. Herbicides and pyraclostrobin (V4) + pyraclostrobin+ epoxiconazole (R2 and R5.1), 4. Herbicides and pyraclostrobin (V4) + epoxiconazole (R2 and R5.1). 5. Herbicides (V4) + pyraclostrobin + epoxiconazole (R2 and R5.1). The herbicides used in MG/BR-46 Conquista were sethoxydim, bentazon and chlorimuron-ethyl; in BRS Valiosa RR, glyphosate was used. The logistic was the model that best adapted to the mean severity ratings and the disease progress curve. At the bottom of canopy, position more favorable to this disease, the logistic model with variable rate was the model that better explain the data. The spraying of strobilurin's group associated with triazole (pyraclostrobin + epoxiconazole) reduced the rate of asian rust development, in relation to the group of ergosterol biosynthesis inhibitors fungicide (epoxiconazole). The pyraclostrobin + epoxiconazole influenced the variables AUDPC, HAA and HAD, and these can be considered suitable to quantify damage caused by this disease. In MG/BR-46 Conquista, the herbicides action affects temporarily the leaf area, affecting indirectly the HAA and HAD variables.