Potential neuroprotective of trans-resveratrol a promising agent tempeh and soybean seed coats-derived against beta-amyloid neurotoxicity on primary culture of nerve cells induced by 2-methoxyethanol / Potencial neuroprotetor do agente trans-resveratrol em cascas de sementes de soja e tempê derivadas da neurotoxicidade beta-amiloide na cultura primária de células nervosas induzidas pelo 2-metoxietanol

Resveratrol, a natural polyphenol found in tempeh, has not been investigated especially in vitro as a neuroprotective agent against 2-methoxyethanol (2-ME)-induced beta-amyloid cytotoxicity. Beta amyloid peptides (Aß) could initiate neurotoxic events and neuron-inflammatory response via microglial activation. However, it remains unknown whether the neurotoxic effect of beta-amyloid and/or associated with the potential of 2-ME to induce neurotoxic effects on primary culture of nerve cells induced by 2-ME. This study investigated potential neuroprotective of trans-resveratrol a promising agent tempeh and soybean seed coats-derived against betaamyloid cytotoxicity on primary culture of nerve cells induced by 2-methoxyethanol. Biotium and MTT assays were used to analyze neurons, which were isolated from the cerebral cortex of fetal mice at gestation day 19 (GD-19). A standard solution of 2-methoxyethanol was dosed at 10 µL. The cultured cells were randomly divided into the following groups: (1) 2-ME group + resveratrol standard, (2) 2-ME group + resveratrol isolated from tempeh, (3) 2-ME group + resveratrol isolated from soybean seed coats, and (4) the control group, without the addition of either 2-ME or resveratrol. Exposure of the primary cortical neuron cells to beta-amyloid monoclonal antibody pre-incubated for 24 h with 10 µL of 4.2 µg/mL resveratrol and 7.5 mmol/l 2-methoxy-ethanol additions. Here, we report that the addition of 2-ME and resveratrol (standard and isolated from tempeh) of cell culture at concentrations of 1.4, 2.8 and 4.2 µg/mL showed that the majority of neurons grew well. In contrast, after exposure to 2-ME and Beta-amyloid, showed that glial activated. These findings demonstrate a role for resveratrol in neuroprotective-neurorescuing action.

O resveratrol, um polifenol natural encontrado em tempê, não foi investigado apenas in vitro como agente neuroprotetor contra a citotoxicidade beta-amiloide induzida por 2-metoxietanol (2-ME). Os peptídeos betaamiloides (Aß) podem iniciar eventos neurotóxicos e resposta inflamatória dos neurônios via ativação microglial. No entanto, permanece desconhecido se o efeito neurotóxico do peptídeo beta-amiloide associado ao potencial do 2-ME causa efeitos neurotóxicos na cultura primária de células nervosas induzidas pelo 2-ME. Este estudo investigou o potencial neuroprotetor do agente trans-resveratrol em cascas de sementes de soja e tempê derivadas da citotoxicidade beta-amiloide na cultura primária de células nervosas induzidas pelo 2-metoxietanol. Ensaios de biotium e MTT foram utilizados para analisar os neurônios isolados do córtex cerebral de camundongos fetais no dia da gestação 19 (GD-19). As células cultivadas foram divididas aleatoriamente nos seguintes grupos: (1) grupo 2-ME + padrão de resveratrol; (2) grupo 2-ME + resveratrol isolado de tempê; (3) grupo 2-ME + resveratrol isolado de cascas de sementes de soja; e (4) grupo controle, sem a adição de 2-ME ou resveratrol. Houve exposição das células primárias dos neurônios corticais ao anticorpo monoclonal beta-amiloide pré-incubado por 24 horas, com 10 µL de 4,2 µg/mL de resveratrol, e adições de 7,5 mmol/l de 2-metoxietanol. A adição de 2-ME e resveratrol (padrão e isolado do tempê) da cultura de células nas concentrações de 1,4, 2,8 e 4,2 µg/mL mostrou que a maioria dos neurônios cresceu bem. Por outro lado, após a exposição ao 2-ME e beta-amiloide, a glia foi ativada. Esses achados demonstram um papel do resveratrol na ação neuroprotetora e de neurorresgate.

Pterostilbeno Reduz o Estresse Oxidativo no Pulmão e no Ventrículo Direito Induzido por Infarto do Miocárdio Experimental / Pterostilbene Reduces Experimental Myocardial Infarction-Induced Oxidative Stress in Lung and Right Ventricle

Resumo Fundamento O pterostilbeno (PS), um composto polifenólico natural e antioxidante, surge como uma intervenção promissora para minimizar danos do infarto agudo do miocárdio (IAM). Objetivo Este estudo teve como objetivo avaliar o desempenho do PS na promoção da homeostase redox nos pulmões e no ventrículo direito (VD) de animais infartados. Métodos Ratos Wistar machos (60 dias de idade) foram randomizados em três grupos: SHAM, IAM (infarto) e IAM+PS (IAM + pterostilbeno). Sete dias após o procedimento de IAM, os ratos foram tratados com PS (100 mg/kg/dia) por gavagem por oito dias. Os animais foram depois sacrificados e os pulmões e VD foram coletados para análise do balanço redox (diferenças foram consideradas significativas quando p<0,05). Resultados Nossos resultados mostram que o IAM desencadeia a interrupção redox no VD e nos pulmões, o que pode contribuir para danos induzido pelo IAM nesses órgãos. Consistentemente, o PS mitigou o estresse oxidativo e restaurou as defesas antioxidantes (Glutationa - GSH nos pulmões: SHAM = 0,79 ± 0,07; IAM = 0,67 ± 0,05; IAM + PS = 0,86 ± 0,14; p<0,05), indicando seu papel protetor neste cenário. Conclusão Nosso trabalho evidencia o potencial do uso de PS como abordagem terapêutica adjuvante após IAM para proteção dos tecidos pulmonares e cardíacos direitos.

Abstract Background Pterostilbene (PS), a natural and antioxidant polyphenolic compound emerges as a promising intervention in improving the myocardial infarction (MI) damages. Objetives This study aimed to evaluate PS actions in promoting redox homeostasis in lungs and right ventricle (RV) of infarcted animals. Methods Male Wistar rats (60 day-old) were randomized into three groups: SHAM, MI (infarcted), and MI+PS (MI+pterostilbene). Seven days after MI procedure, rats were treated with PS (100 mg/kg/day) via gavage for eight days. Animals were euthanized and the lungs and RV were harvested for analyses of redox balance (Differences were considered significant when p<0.05). Results Our results show that MI triggers a redox disruption scenario in RV and lungs, which can contribute to MI-induced damage on these organs. Consistently, PS mitigated oxidative stress and restored antioxidant defenses (GSH in lungs: SHAM= 0.79±0.07; MI=0.67±0.05; MI+PS=0.86±0.14; p<0.05), indicating its protective role in this scenario. Conclusions Our work evidences the PS potential use as an adjuvant therapeutic approach after MI focusing on protecting pulmonary and right-sided heart tissues.

Cardioprotection stimulated by resveratrol and grape products prevents lethal cardiac arrhythmias in an animal model of ischemia and reperfusion

ABSTRACT Purpose To evaluate the preventive cardioprotective effects of resveratrol and grape products, such as grape juice and red wine, in animal model of cardiac ischemia and reperfusion. Methods Male Wistar rats orally pretreated for 21-days with resveratrol and grape products were anesthetized and placed on mechanical ventilation to surgically induce cardiac ischemia and reperfusion by obstruction (ischemia) followed by liberation (reperfusion) of blood circulation in left descending coronary artery. These rats were submitted to the electrocardiogram (ECG) analysis to evaluate the effects of pretreatment with resveratrol and grape products on the incidence of ventricular arrhythmias (VA), atrioventricular block (AVB) and lethality (LET) resulting from cardiac ischemia and reperfusion. Results It was observed that the incidence of AVB was significantly lower in rats pretreated with resveratrol (25%), grape juice (37.5%) or red wine (12.5%) than in rats treated with saline solution (80%) or ethanol (80%). Similarly, incidence of LET was also significantly lower in rats pretreated with resveratrol (25%), grape juice (25%) or red wine (0%) than in rats treated with saline solution (62.5%) or ethanol (75%). Conclusions These results indicate that the cardioprotective response stimulated by resveratrol and grape products prevents the lethal cardiac arrhythmias in animal model of ischemia and reperfusion, supporting the idea that this treatment can be beneficial for prevention of severe cardiac arrhythmias in patients with ischemic heart disease.

Sorting and cryopreservation of goat sperm with or without phenolic compounds / Seleção e criopreservação de espermatozoides caprinos com ou sem compostos fenólicos

The objectives of this study were to evaluate goat sperm sorting in continuous Percoll® density gradients and gamete freezability, in the presence or absence of phenolic antioxidants. For this, semen pools were sorted, frozen, and evaluated. The non-selected group (NSg) presented lower progressive motility (PM), linearity (LIN), straightness (STR), and wobble (WOB) than the selected groups, and straight line velocity (VSL) compared to those with catechin or resveratrol. The amplitude of lateral head displacement (ALH) was higher in NSg, and quercetin reduced the mitochondrial membrane potential (MMP). After thawing, the NSg presented lower PM than the selected groups, VSL and VAP (average path velocity) than the selected group with or without catechin, LIN and WOB than the selected with or without catechin or resveratrol, and STR than the selected with catechin. Moreover, NSg presented higher ALH and BCF than the samples selected with or without catechin. Plasma membrane integrity and intact and living cells were higher in the selected groups, and MMP was lower in the NSg and the selected group with quercetin. Thus, centrifugation in Percoll® continuous density gradients is a viable methodology to select goat sperm compatible with the freezing, especially in the presence of catechin or resveratrol.(AU)

Objetivou-se avaliar a separação de espermatozoides caprinos em gradientes de densidade contínuos de Percoll® e a congelabilidade espermática, com ou sem antioxidantes fenólicos. Para tal, pools seminais foram selecionados, congelados e avaliados. O grupo não selecionado (gNS) apresentou menor motilidade progressiva (MP), linearidade (LIN), retilinearidade (STR) e oscilação (WOB) do que os selecionados, bem como menor velocidade linear progressiva (VSL) do que os com catequina ou resveratrol. A amplitude de deslocamento lateral de cabeça (ALH) foi maior no gNS e a quercetina reduziu o potencial de membrana mitocondrial (PMM). Após a descongelação, o gNS manifestou menor MP do que os selecionados, menor VSL e VAP (velocidade média da trajetória) do que os com ou sem catequina, menor LIN e WOB do que os com ou sem catequina ou resveratrol, e menor STR do que os com catequina, além de maior ALH e BCF do que os com ou sem catequina. A integridade da membrana plasmática e as células intactas e vivas foram maiores nas amostras selecionadas e o PMM foi inferior no gNS e no selecionado com quercetina. Portanto, a centrifugação em gradientes contínuos de densidade de Percoll® é uma metodologia viável para selecionar espermatozoides caprinos compatíveis com a congelação, especialmente na presença de catequina ou resveratrol.(AU)

Polydatin protects H9c2 cells from hypoxia-induced injury via up-regulating long non-coding RNA DGCR5

Polydatin (PD), a monocrystalline polyphenolic drug mainly found in the roots of Polygonum cuspidatum, has various pharmacological activities. Long non-coding RNAs (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) was found to participate in the suppression of multiple cancers. Here, we proposed to study the effect of PD on myocardial infarction (MI) by inducing DGCR5. CCK-8 assay was performed to detect the viability of H9c2 cells. Flow cytometry was utilized to test apoptosis of H9c2 cells. These results determined the optimal concentration and effect time of hypoxia as well as PD. Si-DGCR5 was transfected into cells and the expression level was determined by qRT-PCR. Western blot was utilized to evaluate the expression of apoptosis-related proteins, Bcl-2, Bax, and cleaved-caspase-3, as well as autophagy-associated proteins including Beclin-1, p62, and LC3-II/LC3-I. As a result, PD efficiently attenuated hypoxia-induced apoptosis and autophagy in H9c2 cells. The expression of DGCR5 was down-regulated by hypoxia and up-regulated by PD. Besides, knocking-down the expression of DGCR5 inhibited the protection of PD in H9c2 cells. In addition, PD up-regulated the accumulation of DGCR5, DGCR5 decreased the expression of Bcl-2 and p62, raised the expression of Bax and cleaved-caspase-3, and the proportion of LC3-II/LC3-I. PD stimulated the PI3K/AKT/mTOR and MEK/ERK signaling pathways via up-regulating the expression of DGCR5. Our data demonstrated that PD reduced cell apoptosis and autophagy induced by hypoxia in cardiomyocytes. Moreover, PD activated PI3K/AKT/mTOR and MEK/ERK signaling pathways by up-regulating the expression of DGCR5.

Systemic supplement with resveratrol increased bone formation in rats' alveolar socket / Aumento de la formación ósea en el hueso alveolar de rata con suplemento sistémico con resveratrol

Resveratrol in cell culture media increases osteoblastic markers. Also results from previous studies provide evidence for resveratrol positive effects on bone healing and bone production. In this preclinical study we investigated bone healing in rats by resveratrol systemic application. 30 Wistar male rats were divided into two groups (study group and control group). At first, maxillary second molars of rats were extracted. The rats were kept in laboratory for next 28 days. Study group received resveratrol 20 mg/kg by abdominal injection every day. The control group received placebo in the same manner that study group. Rats were sacrificed after 28 days and bone samples were collected from center of maxillary second molar socket. Samples were evaluated histologically for new bone formation, inflammation, necrosis, fibrosis and foreign body reaction. The mean difference of new bone formation in control group (28.30 %) and study group (45 %) were statistically significant (P=0.014). There were no significant differences in inflammation, fibrosis, necrosis and foreign body reaction (P>0.05). Resveratrol has positive effects on bone healing but more evidence needed from more clinical and animal studies.

El resveratrol en los medios de cultivo celular aumenta los marcadores osteoblásticos. Los resultados de estudios anteriores proporcionan evidencia de efectos positivos del resveratrol sobre la curación ósea y la producción ósea. En este estudio preclínico, investigamos la curación ósea en ratas mediante la aplicación sistémica de resveratrol. Se dividieron 30 ratas macho Wistar en dos grupos (estudio y control). Inicialmente se extrajeron los segundos molares maxilares de las ratas y los animales se mantuvieron en el laboratorio durante los siguientes 28 días. El grupo de estudio recibió todos los días resveratrol 20 mg/kg por inyección abdominal . El grupo control recibió placebo de la misma manera que el grupo estudio. Las ratas fueron sacrificadas después de 28 días y se recogieron muestras de hueso del centro del segundo molar maxilar. Las muestras se evaluaron histológicamente para la formación de hueso nuevo, inflamación, necrosis, fibrosis y reacción de cuerpo extraño. La media de formación de hueso nuevo en el grupo control (28,30 %) y en el grupo estudio (45 %) fueron estadísticamente significativas (P=0,014). No hubo diferencias significativas en la inflamación, fibrosis, necrosis y reacción al cuerpo extraño (P>0,05). El resveratrol tiene efectos positivos sobre la curación de los huesos, pero aún es necesario realizar más pruebas de estudios clínicos, como también en animales.

Neuroprotective potential of polydatin against motor abnormalities and dopaminergic neuronal loss in rotenone induced Parkinson model / Potencial neuroprotector de la polidatina contra las anomalías motoras y la pérdida neuronal dopaminérgica en un modelo de Parkinson inducido por la rotenona

Among the neurodegenerative disorders, Parkinson disease (PD) is ranked as second most common. The pathological hallmark is selective degeneration of the dopaminergic neurons in the nigro-striatal regions of brain with appearance of the Lewy bodies. Present study explores the neuro-protective potential of polydatin in terms of amelioration of degeneration of dopaminergic neurons in nigro-striatal regions of brain and distorted neuromotor behavior in the rotenone model of Parkinson's disease. Thirty-six male Sprague Dawley rats were divided into three groups. Group A (control), Group B (rotenone treated) and Group C (rotenone+polydatin treated). Rotenone was administrated intraperitoneally (i.p) at a dose of 3 mg/kg/body weight while polydatin was given i.p. at a dose of 50 mg/ kg/body weight for four weeks. Then, animals were sacrificed; substantia nigra (SN) & striatum isolated from brain and five micron thick sections were prepared. Cresyl violet (CV), H&E and Immuno-histochemical staining using anti-TH antibody was done. Motor behavior was assessed weekly throughout the experiment using five different methods. Rotenone treated parkinsonian animals showed deterioration of motor behavior, weight loss, loss of dopaminergic neurons and diminished immune-reactivity in the sections from the nigrostriatal regions of these animals Polydatin+rotenone treatment showed contradicting effects to parkinsonism, with amelioration in weight loss, neuro-motor behavior, dopaminergic loss and immune-reactivity against dopaminergic neurons. Present study revealed a neuro-protective potential of polydatin in animal model of PD by ameliorating the neuro-motor abnormalities and degeneration of dopaminergic neurons in nigrostriatal regions.

Entre los trastornos neurodegenerativos, la enfermedad de Parkinson (EP) se clasifica como la segunda más común. El sello patológico es la degeneración selectiva de las neuronas dopaminérgicas en las regiones nigro-estriatales del cerebro, con la aparición de los cuerpos de Lewy. El presente estudio explora el potencial de protección neuronal de la polidatina en términos de la mejora de la degeneración de las neuronas dopaminérgicas en las regiones nigro-estriatales del cerebro y el comportamiento neuromotor distorsionado en el modelo de rotenona de la enfermedad de Parkinson. Treinta y seis ratas macho Sprague Dawley se dividieron en tres grupos: Grupo A (control), Grupo B (tratado con rotenona) y Grupo C (tratamiento con rotenona + polidatina). La rotenona se administró por vía intraperitoneal (i.p.) a una dosis de 3 mg/kg/peso corporal, mientras que la polidatina se administró i.p. a una dosis de 50 mg/kg/ peso corporal durante cuatro semanas. Posteriormente, los animales fueron sacrificados. Se aislaron la substantia nigra (SN) y cuerpo estriado de los cerebros y se realizaron secciones de cinco micras de espesor. Se realizó una tinción de violeta de cresilo (CV), H&E y tinción inmunohistoquímica usando anticuerpo anti-TH. El comportamiento motriz se evaluó semanalmente durante todo el experimento utilizando cinco métodos diferentes. Los animales parkinsonianos tratados con rotenona mostraron deterioro del comportamiento motriz, pérdida de peso, pérdida de neuronas dopaminérgicas y disminución de la reactividad inmune en las secciones de las regiones nigroestriadas. El tratamiento con polidatina + rotenona mostró efectos contrarios al parkinsonismo, con mejoría en la pérdida de peso, en el comportamiento motor, en la pérdida dopaminérgica y en la reactividad inmune contra las neuronas dopaminérgicas. El presente estudio reveló un potencial de protección neuronal de la polidatina en el modelo animal de la EP al mejorar las anomalías neuro-motoras y la degeneración de las neuronas dopaminérgicas en las regiones nigroestriatales.

Increasing pinosylvin production in Escherichia coli by reducing the expression level of the gene fabI-encoded enoyl-acyl carrier protein reductase

Background: The plant secondary metabolite pinosylvin is a polyphenol from the stilbene family, which have positive effects on human health. Biotechnological production is an attractive alternative for obtaining this stilbene. In Escherichia coli, malonyl-CoA is the precursor for both stilbene and fatty acid syntheses. In this study, with the aim of increasing pinosylvin production, we evaluated a novel approach that is based on reducing the expression of the gene fabI, which encodes the enzyme enoyl-acyl carrier protein reductase that is involved in fatty acid synthesis. Results: A recombineering method was employed to eliminate the chromosomal -35 promoter sequence and the upstream region of the gene fabI in E. coli strain W3110. Analysis, employing RT-qPCR, showed that such modification caused a 60% reduction in the fabI transcript level in the mutant strain W3110Δ-35fabI::Cm compared to the wild type W3110. Synthetic genes encoding a mutant version of 4-coumaroyl-CoA ligase from Streptomyces coelicolor A3 with improved catalytic activity employing cinnamic acid as substrate and a stilbene synthase from Vitis vinifera were cloned to generate the plasmid pTrc-Sc4CL(M)-VvSTS. The production performance of strains W3110Δ-35fabI::Cm/pTrc-Sc4CL(M)-VvSTS and W3110/pTrc-Sc4CL(M)- VvSTS was determined in shake flask cultures with Luria-Bertani medium supplemented with 10 g/L glycerol and 3 mM cinnamic acid. Under these conditions, the strain W3110Δ-35fabI::Cm/pTrc-Sc4CL(M)-VvSTS produced 52.67 mg/L pinosylvin, a level 1.5-fold higher than that observed with W3110/pTrc-Sc4CL(M)-VvSTS. Conclusion: A reduction in the transcript level of fabI caused by the elimination of the -35 and upstream promoter sequences is a successful strategy to improve pinosylvin production in E. coli.

Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma

Polydatin, a small molecule from Polygonum cuspidatum, has many biological functions, particularly anti-cancer effects. However, the anti-cancer effects of polydatin in hepatocellular carcinoma (HCC) have not been examined yet. In the present study, MTT assay, BrdU assay, transwell invasion assay, and wound healing assay were performed to determine cell proliferation, invasion and migration. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Quantitative real-time PCR and western blotting assays were used to determine mRNA and protein expression levels. Xenograft experiment was performed to determine the in vivo anti-tumor effect of polydatin. Immunostaining was performed to analyze the expression of caspase-3 and Ki-67. Our results showed that polydatin inhibited cell proliferation in a concentration-dependent and time-dependent manner in the HCC cell lines. Polydatin also induced cell apoptosis in a concentration-dependent manner possibly via increasing the caspase-3 activity, and up-regulating the protein expression of caspase-3, caspase-9, Bax, and down-regulating the protein expression of Bcl-2. In addition, polydatin treatment had an inhibitory effect on cell proliferation, invasion and migration in HCC cell lines. Polydatin treatment also suppressed the Wnt/beta-catenin signaling activities in HCC cells. Polydatin treatment significantly reduced tumor growth in nude mice inoculated with HepG2 cells, suppressed the expression of Ki-67, and increased caspase-3 expression and TUNEL activity. Our data indicated the important role of polydatin for the suppression of HCC progression.

Therapeutic effect of resveratrol on morphine-induced damage in male reproductive system of mice by reducing nitric oxide serum level / Efecto terapéutico de resveratrol sobre el daño inducido por la morfina en el sistema reproductor masculino de ratones reduciendo el nivel de óxido nítrico del suero

SUMMARY: Morphine is one of the naturally occurring phenanthrene alkaloids of opium that induces adverse effects on male reproductive system. Resveratrol is a phytoestrogen and antioxidant of red grape. The main goal is to investigate whether resveratrol could inhibit adverse effects of morphine on sperm cell viability, count, motility as well as testis histology, testosterone hormone and nitric oxide levels in mice. In the present study, 48 male rats were randomly divided into 8 groups (n=6) and were treated intraperitoneally for 14 days with normal saline, resveratrol (2, 8, 20 mg/kg/day), morphine (20 mg/kg/day) and morphine (20 mg/kg/day) + resveratrol (2, 8, 20 mg/kg/day). At the end of experiments, sperm parameters (sperm cell viability, count, motility and morphology), testis weight, the diameter of seminiferous tubules, testosterone hormone level and nitric oxide were analyzed. The data were analyzed by SPSS software for windows (version 20) using one-way ANOVA test followed by Tukey's post hoc test, and P<0.05 was considered significant. The results indicated that morphine administration significantly decreased testosterone level, count, viability and motility of sperm cells and testis weight and increased nitric oxide compared to the saline group (P=0.000). Administration of resveratrol and resveratrol plus morphine significantly increased motility, count and viability of sperm cells, somniferous tubule diameter and testosterone, while it decreased nitric oxide level compared to morphine group (P=0.025). It seems that resveratrol administration could increase the quality of spermatozoa and prevented morphine-induced adverse effects on sperm parameters.

RESUMEN: La morfina es uno de los alcaloides fenantreno del opio que induce efectos adversos en el sistema reproductivo masculino. El resveratrol es un fitoestrógeno y antioxidante de la uva roja. El objetivo principal de este trabajo fue investigar si el resveratrol puede inhibir los efectos adversos de la morfina sobre la viabilidad celular de los espermatozoides, el recuento y la motilidad, así como la histología de los testículos, la hormona testosterona y los niveles de óxido nítrico en ratones. Se dividieron, aleatoriamente, 48 ratas machos en 8 grupos (n = 6) y se trataron de forma intraperitoneal durante 14 días con solución salina normal, resveratrol (2, 8, 20 mg / kg / día), morfina (20 mg / kg / día ) y morfina (20 mg / kg / día) + resveratrol (2, 8, 20 mg / kg / día). Al final de los experimentos, se analizaron los parámetros espermáticos (viabilidad celular, recuento, motilidad y morfología), el peso de los testículos, el diámetro de los túbulos seminíferos, el nivel de la hormona testosterona y el óxido nítrico. Los datos fueron analizados con el software de SPSS para Windows (versión 20) usando una prueba de ANOVA de una vía seguida de la prueba post hoc de Tukey, y P <0,05 se consideró significativo. Los resultados indicaron que la administración de morfina disminuyó significativamente el nivel de testosterona, el recuento, la viabilidad y la motilidad de los espermatozoides y el peso de los testículos, además del aumento de óxido nítrico en comparación con el grupo salino (p = 0,000). La administración de resveratrol y resveratrol más morfina aumentó significativamente la motilidad, el recuento y la viabilidad de los espermatozoides, el diámetro de los túbulos seminíferos y la testosterona, mientras que disminuyó el nivel de óxido nítrico comparado con el grupo morfina (p = 0,025). En conclusión, la administración de resveratrol podría aumentar la calidad de los espermatozoides y prevenir los efectos adversos inducidos por la morfina sobre los parámetros espermáticos.

Protective effect of resveratrol against morphine damage to kidneys of mice / Efecto protector de resveratrol contra el daño de la morfina en riñones de ratones

SUMMARY: Morphine produces free radicals and cause apoptosis in some cell. Resveratrol (RSV) is a stilbenoid, a type of natural phenol, and a phytoalexin produced by several plants in response to injury. 48 male mice were randomly assigned to 8 groups. In this study, various doses of RSV (2, 8 and 20 mg/kg) and RSV plus Morphine (2, 8 and 20 mg/kg) were administered intraperitoneally to male mice for 20 consequent days and weight of kidneys, biochemical characteristics, morphometric markers and blood serum nitric oxide level were studied. The results indicated that morphine administration significantly increased the mean diameter of glomerulus and distal and proximal convoluted tubule, Lactate dehydrogenase (LDH), Blood urea nitrogen (BUN), creatinine and nitric oxide levels compared to the saline group (P<0.05). However, RSV and RSV plus morphine in all doses significantly decreased glomeruli number and LDH, BUN, creatinine and nitric oxide levels compared to morphine groups (p<0.05). Thus, it seems that resveratrol improved kidney damages induced by morphine in mice.

RESUMEN: La morfina produce radicales libres y causa apoptosis en algunas células. El resveratrol (RSV) es un tipo de fenol natural y una fitoalexina producida por varias plantas en respuesta a una lesión. Se asignaron al azar 48 ratones machos a 8 grupos. En este estudio se administraron varias dosis de RSV (2, 8 y 20 mg/kg) y RSV más morfina (2, 8 y 20 mg/kg) intraperitoneal en ratones machos durante 20 días consecutivos y se estudió el peso de los riñones, las características bioquímicas, los marcadores morfométricos y el nivel de óxido nítrico en suero sanguíneo. Los resultados indicaron que la administración de morfina aumentó significativamente el diámetro medio del glomérulo y de los túbulos distal y proximal, los niveles de lactato deshidrogenasa (LDH), nitrógeno ureico en sangre (BUN), la creatinina y el óxido nítrico en comparación con el grupo salino (p <0,05). Sin embargo, el RSV y el RSV más morfina en todas las dosis redujeron significativamente el número de glomérulos y LDH, BUN, la creatinina y el óxido nítrico en comparación con los grupos de morfina (p <0,05). Por lo tanto, los resultados podrían indicar que el resveratrol mejoró el daño renal inducido por la morfina en ratones.

Lycopene and resveratrol pretreatment did not interfere with the liver of hepatectomized rats

Abstract Purpose: To investigate the effects of lycopene and resveratrol pretreatment on hepatic hyperplasia in partially hepatectomized rats. Methods: The lycopene group and the resveratrol group received 40 mg/kg/day of lycopene or resveratrol, respectively (dissolved in olive oil or in saline solution, respectively) and administered via a gastric tube for 30 days. The partially hepatectomzed (PH) control groups received saline or olive oil via a gastric tube for 30 days, respectively, and the normal control group received no treatment. Liver tissue and intracardiac blood samples were obtained 24, 36 or 48 h after PH. Results: No areas of fibrosis were detected. No significant changes in mitotic index, in the number of apoptosis events or in aspartate aminotransferase and alanine aminotransferase levels were observed. Conclusions: Lycopene and resveratrol pretreatment did not interfere on hepatic hyperplasia in partially hepatectomized rats.

Resveratrol Treatment Normalizes the Endothelial Function and Blood Pressure in Ovariectomized Rats / Tratamento com Resveratrol Normaliza a Função Endotelial e a Pressão Arterial em Ratas Ovariectomizadas

Abstract Background: Despite knowing that resveratrol has effects on blood vessels, blood pressure and that phytostrogens can also improve the endothelium-dependent relaxation/vasodilation, there are no reports of reveratrol's direct effect on the endothelial function and blood pressure of animals with estrogen deficit (mimicking post-menopausal increased blood pressure). Objective: To verify the effect of two different periods of preventive treatment with resveratrol on blood pressure and endothelial function in ovariectomized young adult rats. Methods: 3-month old female Wistar rats were used and distributed in 6 groups: intact groups with 60 or 90 days, ovariectomized groups with 60 or 90 days, and ovariectomized treated with resveratrol (10 mg/kg of body weight per day) for 60 or 90 days. The number of days in each group corresponds to the duration of the experimental period. Vascular reactivity study was performed in abdominal aortic rings, systolic blood pressure was measured and serum nitric oxide (NO) concentration was quantified. Results: Ovariectomy induced blood pressure increase 60 and 90 days after surgery, whereas the endothelial function decreased only 90 days after surgery, with no difference in NO concentration among the groups. Only longer treatment (90 days) with resveratrol was able to improve the endothelial function and normalize blood pressure. Conclusion: Our results suggest that 90 days of treatment with resveratrol is able to improve the endothelial function and decrease blood pressure in ovariectomized rats.

Resumo Fundamentos: Apesar de se saber que o resveratrol apresenta efeitos sobre a pressão arterial e os vasos sanguíneos, e que os fitoestrógenos podem melhorar o relaxamento/vasodilatação dependente do endotélio, não há relatos do efeito direto do resveratrol sobre a pressão arterial e a função endotelial em animais com deficiência de estrógeno (mimetizando a pressão arterial aumentada pós-menopausa). Objetivo: Verificar o efeito de dois diferentes períodos de tratamento preventivo com resveratrol sobre a pressão arterial e a função endotelial em ratas adultas jovens ovariectomizadas. Métodos: Foram utilizadas ratas Wistar com 3 meses de idade, distribuídas em 6 grupos: grupos intactas com 60 ou 90 dias, grupos ovariectomizadas com 60 ou 90 dias, grupos ovariectomizadas e tratadas com resveratrol na dose de 10mg/kg de massa corporal por dia, durante 60 ou 90 dias, sendo o número de dias em cada grupo relativo à duração do período experimental. Foi realizado um estudo de reatividade vascular em anéis da aorta abdominal, mensurada a pressão arterial sistólica e quantificada a concentração sérica de óxido nítrico (NO). Resultados: A ovariectomia induziu aumento da pressão arterial 60 e 90 dias após a cirurgia, enquanto a função endotelial decaiu apenas após 90 dias, e não houve diferença na concentração de NO entre os grupos. Apenas o tratamento prolongado com resveratrol (90 dias) foi capaz de melhorar a função endotelial e normalizar a pressão arterial. Conclusão: Nossos resultados sugerem que o tratamento por 90 dias com resveratrol é capaz de melhorar a função endotelial e diminuir a pressão sanguínea em ratas ovariectomizadas.

High temperature and UV-C treatments affect stilbenoid accumulation and related gene expression levels in Gnetum parvifolium

Background: Gnetum parvifolium stems and roots have been used for a long time in traditional Chinese medicines. Stilbenes are bioactive compounds present in G. parvifolium plants, and they possess antioxidative and anticancer properties. However, little is known about the responses of G. parvifolium stilbene biosynthetic pathways to stress conditions. Therefore, we investigated stilbene biosynthesis, including the expression of relevant genes, in G. parvifolium exposed to high-temperature and ultraviolet-C treatments. Results: High temperatures did not influence the accumulation of total stilbenes in stems but decreased stilbene concentrations in roots at 3 h, with a subsequent restoration to control levels. In contrast, ultraviolet irradiation induced the accumulation of total stilbenes in stems but not in roots. We also observed that high temperatures inhibited the production of resveratrol and piceatannol in G. parvifolium stems and roots, whereas ultraviolet treatments initially inhibited their accumulation (up to 6 h) but induced their production at later time points. Analyses of specific genes (i.e., PAL, C4H, 4CL, STS, and CYP) revealed that their expression levels generally increased in stress-treated stems and roots, although there was some variability in the expression profiles during treatments. Conclusions: Our results indicated that high temperatures and ultraviolet irradiation differentially affect the biosynthesis of specific stilbenes in G. parvifolium stems and roots. Therefore, cultivating G. parvifolium seedlings under optimal stress conditions may increase the biosynthesis of specific stilbene compounds.

Effect of different presentations of resveratrol on cell proliferation and epitelial thickness of the oral mucosa of wistar rats

Introduction: Grape is one of the most important fruit crops across the world and can be consumed in different ways. There has been a growing interest in the role of antioxidants such as resveratrol, which can be found in grape skin, in oral and dental tissues. Thus, the objective of this study was to analyze the effect of different presentations of resveratrol on cell proliferation and epithelial thickness of the oral mucosa of Wistar rats. Methods: Fifty male Wistar rats were randomly divided into five groups: water/control, red wine, grape juice, 12% alcoholic solution/ethanol and aqueous solution of resveratrol. Samples of palatal and tongue mucosa were collected for a histomorphometric analysis using hematoxylin-eosin staining and the argyrophilic nucleolar organizer region (AgNOR) technique for quantification of cell proliferation. Results: As to epithelial thickness, both the tongue and the palate showed a statistically significant difference between the control group and the other groups, with greater decrease in the resveratrol and the wine groups. In the suprabasal layer of both the tongue and the palate epithelium, red wine reduced the rate of cell proliferation, while ethanol increased it. In the basal layer of the tongue epithelium, there was a statistically significant difference between the control, the grape juice and the resveratrol groups and the ethanol group, with increased cell proliferation in the ethanol group. Conclusions: Wine does not interfere in the physiological renewal of the basal layer of the buccal epithelium and exerts a protective action by reducing the cell proliferation rate of the suprabasal layer (AU)

Piceatannol induced apoptosis through up-regulation of microRNA-181a in melanoma cells

BACKGROUND: Melanoma took top position among the lethal cancers and, despite there have been some great attempts made to increase the natural life of patients with metastatic disease, long-lasting and complete remissions are few. Piceatannol, owns the similar function as resveratrol, has been defined as an anti-cancer agent playing important role in inhibition of proliferation, migration and metastasis in various cancer. Thus, we aim to investigate the anti-cancer effect and mechanisms of piceatannol in melanoma cells. METHODS: Melanoma cell lines WM266-4 and A2058 were treated either with or without piceatannol. Cell viability and cell apoptosis were assessed by using MTT and Annexin V/PI assay, respectively. Cells were transfected with specific miRNA using Lipfectamine 2000. miRNA bingding ability to 3'-UTR region within specific gene was assed by firefly luciferase analysis. Gene and protein expression was eveluated by qRT-PCR and western blot analysis, respectively. RESULTS: Our study showed that piceatannol inhibited WM266-4 and A2058 cells growth and induced apoptosis. Totally, 16 differentially expressed miRNAs were screened out including 8 up-regulated and 8 down-regulated miRNAs. Expression level of miR-181a is significantly higher in piceatannol-treated cells than normal control and is lower in melanoma cancer tissues than its adjacent normal tissues. Bcl-2 is a target gene of miR-181a. Moreover, silencing of miR-181a reverses the decrease of cell viability induced by piceatannol in WM266-4 and A2058 cells. Taken together, present study uncovered the ability of piceatannol to repress melanoma cell growth and clarified the contribution of miR-181a in the anticancer role of piceatannol. CONCLUSION: The present study proposes that piceatannol can be taken into account to be a hopeful anticancer agent for melanoma.

Effect of Resveratrol on periodontal pathogens during experimental periodontitis in rats

Abstract The objective of this study was to investigate the antibacterial effect of resveratrol against putative periodontal pathogens during the progression of experimental periodontitis in rats. Periodontitis was induced in rats in one of the first molars chosen to receive a ligature. Animals were assigned to one of two groups: daily administration of the placebo solution (control group, n = 12) or 10 mg/Kg of resveratrol (RESV group, n = 12). The therapies were administered systemically for 30 days, for 19 days before periodontitis induction and then for another 11 days. Then, the presence and concentrations of Porphyromonas gingivalis, Tannerella forsythia and Aggregatibacter actinomycetemcomitans in the cotton ligatures collected from the first molars were evaluated using real-time PCR. Inter-group comparisons of the microbiological outcomes revealed that no differences were detected for P. gingivalis, T. forsythia and A. actinomycetemcomitans levels (p > 0.05). Continuous use of resveratrol did not promote additional benefits in microbiological outcomes during experimental periodontitis in rats.

Micro-CT evaluation of the radioprotective effect of resveratrol on the mandibular incisors of irradiated rats

Abstract The purpose of this study was to perform a microcomputed tomographic evaluation of the radioprotective effect of resveratrol on the volume of mandibular incisors of irradiated rats. A second aim was to make a quantitative assessment of the effect of x-ray exposure on these dental tissues. Twenty adult male rats were divided into four groups: control, irradiated control, resveratrol, and irradiated resveratrol. The resveratrol groups received 100 mg/kg of resveratrol, whereas the irradiated groups were exposed to 15 Gy of irradiation. The animals were sacrificed 30 days after the irradiation procedure, and their mandibles were removed and scanned in a microcomputed tomography unit. The images were loaded into Mimics software to allow segmentation of the mandibular incisor and assessment of its volume. The results were compared by One-way ANOVA and Tukey's post hoc test, considering a 5% significance level. The irradiated groups showed significantly diminished volumes of the evaluated teeth, as compared with the control group (p < 0.05). The resveratrol group presented higher values than those of the irradiated groups, and volumes similar to those of the control group. High radiation doses significantly affected tooth formation, resulting in alterations in the dental structure, and thus lower volumes. Moreover, resveratrol showed no effective radioprotective impact on dental tissues. Future studies are needed to evaluate different concentrations of this substance, in an endeavor to verify its potential as a radioprotector for these dental tissues.

Effect of resveratrol on mitochondrial activity in differentiated mature adipocytes / Efecto del resveratrol sobre la actividad mitocondrial en adipocitos maduros diferenciados

Obesity has reached epidemic proportions, the World Health Organization (WHO) estimates that there are more than 1,000 million overweight adults world-wide. Furthermore, obesity is characterized as an overgrowth of white adipose tissue as a result of adipocyte hypertrophy and hyperplasia. Mitochondria is considered the source of energy within the adipocyte, since it contains the molecular machinery, and it is involved in a large number of metabolic pathways, besides the transformation of chemical energy into adenosine triphosphate. Mitochondria shortage and adipocyte dysfunction result in an excessive accumulation of triacylglycerol in the cytoplasm, which determines an imbalance between energy production and energy expenditure. Resveratrol (RSV) is a polyphenol found in different plants and its effects have been associated with mitochondrial biogenesis. An adipogenesis in vitro model (3T3-L1 preadipocytes) was used, and these cells were differentiated into mature adipocytes. Subsequently the effect of RSV on the adipocytes morphology, the lipid content and mitochondrial activity was evaluated using microscopic and flow cytometry techniques. The effect of RSV on differentiated mature adipocytes, was characterized by the decrease in lipid content and the consequently declination of the mitochondrial activity. 3T3-L1 preadipocytes retained the differentiation ability until passage 18. The RSV at doses of 25 and 50 µM for 48 hours in differentiated mature adipocytes promoted the decreased in lipid content probably due to an increase in mitochondrial activity in the early hours of RSV exposure, causing the consequently declination of mitochondrial activity at the end of 48 hours.

La obesidad ha tomado dimensiones epidémicas globales y la Organización Mundial de la Salud estima que hay más de 1,000 millones de adultos con sobrepeso. Así mismo, la obesidad se ha caracterizado como la expansión del tejido adiposo blanco condicionada por la hipertrofia e/o hiperplasia de los adipocitos. La mitocondria es considerada la fuente de energía dentro del adipocito, debido a que contiene la maquinaria molecular que dirige, a través de diversas vías metabólicas, la transformación de la energía química en adenosíntrifosfato. La escasez de mitocondrias así como su disfunción en el adipocito, resulta en una acumulación excesiva de triacilgliceroles en el citoplasma, lo que condiciona un desequilibrio entre producción de energía y gasto energético. El resveratrol (RSV) es un polifenol que se encuentra en diferentes grupos de plantas y sus efectos se han asociado con la inducción de genes para la biogénesis mitocondrial. Se empleó un modelo de adipogénesis (in vitro) materializado por una línea celular de preadipocitos 3T3-L1, mismos que se diferenciaron a adipocitos maduros. Posteriormente se evaluó el efecto del RSV sobre la morfología, contenido lipídico y actividad mitocondrial en los adipocitos maduros diferenciados a través de las técnicas: microscopía invertida, confocal y citometría de flujo. El efecto del RSV sobre los adipocitos maduros diferenciados, se caracterizó por la disminución del contenido lipídico y consecuentemente de la actividad mitocondrial. Los preadipocitos 3T3-L1 conservaron la capacidad de diferenciación hasta el pase 18. Por otra parte, el resveratrol a dosis de 25 y 50 µM durante 48 horas en adipocitos maduros diferenciados, promueve una disminución en el contenido lipídico probablemente debido a un aumento de la actividad mitocondrial en las primeras horas de exposición al tratamiento, provocando la disminución de la actividad mitocondrial al término de 48 horas.