RESUMO
SUMMARY: In this study we aimed to examine the effect of novel vasodilatory drug Riociguat co-administration along resveratrol to recover neurodegeneration in experimental stroke injury. For that purpose, thirty-five adult female rats were divided into five groups (Control, MCAO, MCAO + R, MCAO + BAY, MCAO + C) of seven animals in each. Animals in Control group did not expose to any application during the experiment and sacrificed at the end of the study. Rats in the rest groups exposed to middle cerebral artery occlusion (MCAO) induced ischemic stroke. MCAO + R group received 30 mg/kg resveratrol, and MCAO + BAY group received 10 mg/kg Riociguat. The MCAO + C group received both drugs simultaneously. The drugs were administered just before the reperfusion, and the additional doses were administered 24h, and 48h hours of reperfusion. All animals in this study were sacrificed at the 72nd hour of experiment. Total brains were received for analysis. Results of this experiment indicated that MCAO led to severe injury in cerebral structure. Bax, IL-6 and IL-1ß tissue levels were up-regulated, but anti-apoptotic Bcl-2 immunoexpression was suppressed (p<0.05). In resveratrol and Riociguat treated animals, the neurodegenerations and apoptosis and inflammation associated protein expressions were improved compared to MCAO group, but the most success was obtained in combined treatment exposed animals in MCAO + C group. This study indicated that the novel soluble guanylate stimulator Riociguat is not only a potent neuroprotective drug in MCAO induced stroke, but also synergistic administration of Riociguat along with resveratrol have potential to increase the neuroprotective effect of resveratrol in experimental cerebral stroke exposed rats.
En este estudio, nuestro objetivo fue examinar el efecto de la coadministración del nuevo fármaco vasodilatador Riociguat junto con resveratrol para recuperar la neurodegeneración en lesiones por ataques cerebrovasculares experimentales. Para ello, se dividieron 35 ratas hembras adultas en cinco grupos (Control, MCAO, MCAO + R, MCAO + BAY, MCAO + C) de siete animales en cada uno. Los animales del grupo control no fueron sometidos a ninguna aplicación durante el experimento y se sacrificaron al final del estudio. Las ratas de los grupos expuestas a la oclusión de la arteria cerebral media (MCAO) indujeron un ataque cerebrovascular isquémico. El grupo MCAO + R recibió 30 mg/kg de resveratrol y el grupo MCAO + BAY recibió 10 mg/kg de Riociguat. El grupo MCAO + C recibió ambos fármacos simultáneamente. Los fármacos se administraron antes de la reperfusión y las dosis adicionales se administraron a las 24 y 48 horas de la reperfusión. Todos los animales en este estudio fueron sacrificados a las 72 horas del experimento. Se recibieron cerebros totales para su análisis. Los resultados indicaron que la MCAO provocaba lesiones graves en la estructura cerebral. Los niveles tisulares de Bax, IL-6 e IL- 1ß estaban regulados positivamente, pero se suprimió la inmunoexpresión antiapoptótica de Bcl-2 (p <0,05). En los animales tratados con resveratrol y Riociguat, las neurodegeneraciones y las expresiones de proteínas asociadas a la apoptosis y la inflamación mejoraron en comparación con el grupo MCAO, sin embargo el mayor éxito se obtuvo en el tratamiento combinado de animales expuestos en el grupo MCAO + C. Este estudio indicó que el nuevo estimulador de guanilato ciclasa soluble Riociguat no solo es un fármaco neuroprotector potente en el ataque cerebrovascular inducido por MCAO, sino que también la administración sinérgica de Riociguat junto con resveratrol tiene el potencial para aumentar el efecto neuroprotector del resveratrol en ratas experimentales expuestas a un ataque cerebrovascular.
Assuntos
Animais , Feminino , Ratos , Pirazóis/administração & dosagem , Pirimidinas/administração & dosagem , Acidente Vascular Cerebral/tratamento farmacológico , Resveratrol/administração & dosagem , Arteriopatias Oclusivas , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Interleucina-6/análise , Apoptose/efeitos dos fármacos , Fármacos Neuroprotetores , Artéria Cerebral Média , Acidente Vascular Cerebral/patologia , Ativadores de Enzimas/administração & dosagem , Modelos Animais , Quimioterapia Combinada , Interleucina-1beta/análise , Guanilato Ciclase/efeitos dos fármacos , InflamaçãoRESUMO
SUMMARY: BPA is a multifunctional endocrine disruptor with ubiquitous presence in aquatic ecosystems. The Mexican Central Plateau is an area severely impacted by pollution, inhabited by endemic viviparous fish. However, efforts to understand the effects of BPA on native species such as Goodea atripinnis are non-existent. This study focused on providing in vivo evidence of alterations in the testes of G. atripinnis males due to acute exposure to BPA at test concentrations of 1 mg/L, 10 mg/L, and 50 mg/L for 96 h. BPA exposition 1 mg/L and 10 mg/L showed degeneration and disorganization in germinal tissue. Furthermore, there was a notable decrease in sperm within the seminiferous tubules of males exposed to 10 mg/L of BPA. In all treatments, somatic cells had alterations by connective tissue thickening and an increase in collagen fibers. Additionally, inflammation and bleeding occurred in the testes of males exposed to 1 and 10 mg/L BPA. The alterations in the testes of G. atripinnis are related to BPA toxicity, which can lead to apoptosis in germ cells increasing connective tissue. Finally, even though the changes produced by BPA became evident in acute exposure (96 h), its effects are probably irreversible, compromising the reproduction of G. atripinnis.
El BPA es un disruptor endocrino multifuncional con presencia ubicua en los ecosistemas acuáticos. La Meseta Central mexicana habitada por peces vivíparos endémicos, es una zona severamente impactada por la contaminación. Sin embargo, los esfuerzos por comprender los efectos del BPA en especies nativas como Goodea atripinnis son inexistentes. Este estudio se centró en proporcionar evidencia in vivo de alteraciones en los testículos de machos de G. atripinnis debido a la exposición aguda al BPA en concentraciones de prueba de 1 mg/L, 10 mg/L y 50 mg/L durante 96 h. La exposición a BPA 1 mg/L y 10 mg/L mostró degeneración y desorganización en el tejido germinal. Además, hubo una disminución notable de los espermatozoides dentro de los túbulos seminíferos de machos expuestos a 10 mg/L de BPA. En todos los tratamientos las células somáticas presentaron alteraciones por engrosamiento del tejido conectivo y aumento de las fibras de colágeno. Además, se produjo inflamación y sangrado en los testículos de machos expuestos a 1 y 10 mg/L de BPA. Las alteraciones en los testículos de G. atripinnis están relacionadas con la toxicidad del BPA, lo que puede provocar apoptosis en las células germinales aumentando el tejido conectivo. Finalmente, si bien los cambios producidos por el BPA se hicieron evidentes en la exposición aguda (96 h), sus efectos probablemente sean irreversibles, comprometiendo la reproducción de G. atripinnis.
Assuntos
Animais , Fenóis/toxicidade , Testículo/efeitos dos fármacos , Compostos Benzidrílicos/toxicidade , Ciprinodontiformes , Testículo/patologia , Disruptores Endócrinos , PeixesRESUMO
SUMMARY: Resveratrol (RES) and quercetine (QRC), is a promising agent relevant for both cancer chemoprevention and treatment via several signaling pathways, involved in their anticancer activity related to its chemotherapeutic potential, associated with the induction of ROS generation in cancer cells, leading to apoptosis. In our study, we have summarized the mechanisms of action of RES and QRC, and their pharmacological implications and potential therapeutic applications in cancer therapy. After treatment of Hep 2 cells with QRC or RES, the death pathways such as the cytochrome c release, ERK1/2 and IRS-1 pathways were upregulated, while cell survival pathway, including PI3K/AKT were downregulated. The RES and QRC caused oncosis, cells hypertrophy, hypercondensatin of chromatin, rupture of the plasma membrane and nuclear membrane, and formation of apoptotic bodies. Morphometric measurements of some cellular and nuclear parameters showed that RES and QRC induced an increase in cells and nuclear size, the nucleocytoplasmic ratio remained below 1 (N-Cyt R < 1), sign of low nuclear activity. The RES and QRC induced apoptosis of Hep2 cells by increasing of oxidative stress markers, MDA, and by modulating detoxifying enzymes, CAT and SOD. Our study results prove antiproliferative and proapoptotic properties of quercetin and resveratrol with regard to larynx cancer.
Resveratrol (RES) y quercetina (QRC), es un agente prometedor y relevante tanto para la quimioprevención como para el tratamiento del cáncer a través de varias vías de señalización, involucrado en su actividad anticancerígena relacionada con su potencial quimioterapéutico, asociado con la inducción de la generación de especies reactivas del oxígeno (ROS) en células cancerosas, lo que lleva a apoptosis. En nuestro estudio, hemos resumido los mecanismos de acción de RES y QRC, y sus implicaciones farmacológicas y posibles aplicaciones terapéuticas en la terapia del cáncer. Después del tratamiento de las células Hep 2 con QRC o RES, las vías de muerte, tal como la liberación de citocromo c, las vías ERK1/2 e IRS-1, se regulaban positivamente, mientras que la vía de supervivencia celular, incluida PI3K/AKT, se regulaba negativamente. El RES y el QRC provocaron oncosis, hipertrofia celular, hipercondensación de la cromatina, rotura de la membrana plasmática y nuclear y formación de cuerpos apoptóticos. Las mediciones morfométricas de algunos parámetros celulares y nucleares mostraron que RES y QRC indujeron un aumento en las células y el tamaño nuclear, la proporción nucleocitoplasmática se mantuvo por debajo de 1 (N- Cyt R <1), signo de baja actividad nuclear. RES y QRC indujeron la apoptosis de las células Hep2 aumentando los marcadores de estrés oxidativo, MDA, y modulando las enzimas desintoxicantes, CAT y SOD. Los resultados de nuestro estudio demuestran las propiedades antiproliferativas y proapoptóticas de la quercetina y el resveratrol con respecto al cáncer de laringe.
Assuntos
Humanos , Quercetina/farmacologia , Linhagem Celular Tumoral/efeitos dos fármacos , Resveratrol/farmacologia , Sobrevivência Celular , Morte Celular , Apoptose , Estresse Oxidativo , Proliferação de Células/efeitos dos fármacosRESUMO
Epilepsy is the chronic non-communicable disease of the nervous system most prevalent in the world. Valproic acid (VPA) is one of the most used drugs in the treatment of epilepsy but with various side effects. One of the organs that can be affected is the testis, where it has been seen that men treated with VPA reduce their fertility rates, in addition to causing endocrine disorders by decreasing androgens and gonadotropins. In animal models, it has been shown to reduce the weights of the glands attached to the male reproductive tract, as well as at the testicular level, decreasing sperm concentration and increasing apoptotic cell count. These effects are because VPA increases reactive oxygen species (ROS), causing damage to macromolecules and affecting all cellular processes sensitive to oxide reduction. Throughout testicular development, in utero, it has been seen that the expression of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase, are lower during early embryonic development, as well as vitamin E (VE) is decreased. Therefore, they are not sufficient to reverse the toxic effects of ROS. The objective of this study was to review the use of VPA during pregnancy, its effect on testicular development, and to explore the potential protective role of vitamin E.
La epilepsia es una enfermedad crónica no transmisible que afecta al sistema nervioso más prevalente en el mundo. Dentro de los tratamientos, uno de los fármacos más utilizados es el ácido valproico (AVP), el que ocasiona diversos efectos secundarios. Entre los órganos que se pueden ver afectados se encuentra la gónada masculina, en donde se ha visto que hombres en tratamiento con AVP reducen sus tasas de fecundidad, además de causar trastornos endocrinos disminuyendo andrógenos y gonadotrofinas. En modelos animales, se ha visto que disminuye los pesos de las glándulas anexas al tracto reproductor masculino, como también a nivel testicular, disminuyendo la concentración espermática y aumentando el recuento de células apoptóticas. Estos efectos se deberían a que el AVP aumenta las especies reactivas de oxígeno (ROS), ocasionando daño en macromoléculas, afectando todos los procesos celulares sensibles a óxido reducción. A lo largo del desarrollo testicular, in utero se ha visto que la expresión de enzimas antioxidantes como superóxido dismutasa, catalasa y glutatión peroxidasa, son más bajos durante el desarrollo embrionario temprano, como también la vitamina E (VE) se encuentra disminuida. Por tanto, no resultan suficientes para revertir los efectos tóxicos de las ROS. El objetivo de esta revisión fue asociar el uso de AVP durante la gestación y sus efectos a nivel del desarrollo testicular y describir el potencial rol protector de la VE.
Assuntos
Humanos , Animais , Masculino , Feminino , Gravidez , Testículo/efeitos dos fármacos , Vitamina E/farmacologia , Ácido Valproico/efeitos adversos , Teratogênicos , Testículo/crescimento & desenvolvimento , Ácido Valproico/toxicidade , Espécies Reativas de Oxigênio , Epilepsia/tratamento farmacológico , Desenvolvimento Embrionário e Fetal/efeitos dos fármacosRESUMO
SUMMARY: Underage drinking has become a major public concern having a negative impact on the growth and development of the skeleton. Peak bone mass is attained during adolescence hence the aim of the study was to investigate the effect of acute binge alcohol consumption on trabecular morphometry and tensile strength of the adolescent mandible in the Sprague Dawley (SD) rat. The study comprised of 24 SD rats, aged 7 weeks, placed into either the alcohol-exposed [n=12 (6 males and 6 female)] or pair-fed control group [n=12 (6 male and 6 female)]. The treatment of the groups was as follows; the alcohol exposed group and the pair-fed control were administered a single daily dose of 3 g/kg of 20 % alcohol 3 days a week (alternate days) for 7 days and a caloric equivalent dose of maltose dextrin via oral gavage, respectively. The animals were terminated on day 7 via pentobarbital injection. The mandibles were harvested and scanned using a Nikon XTH 255L 3D-microCT scanner (Nikon Metrology, Leuven, Belgium), and biomechanical tests were done using a Shimadzu universal tensile strength testing machine (China). Following scanning and reconstruction, the trabecular morphometry was assessed using Volume Graphics Studio® software. A 3-point bending test was used to evaluate the tensile strength of the bone. Findings from our study showed changes in some trabecular parameters in the female alcohol-exposed group, while the male groups remained unaffected. No changes in tensile strength were seen when comparing male pair-fed control and alcohol-exposed groups and when comparing female pair-fed control and alcohol-exposed groups. Trabecular and tensile strength differences were observed between the sexes when comparing male pair-fed control and alcohol-exposed groups to female pair-fed control and alcohol-exposed groups. These findings do suggest that acute binge alcohol consumption has detrimental effects on the bone micro-architecture in female alcohol-exposed rats and that differences are seen between the sexes.
El consumo de alcohol entre menores de edad se ha convertido en una importante preocupación pública que tiene un impacto negativo en el crecimiento y desarrollo del esqueleto. La masa ósea máxima se alcanza durante la adolescencia, por lo que el objetivo del estudio fue investigar el efecto del consumo excesivo de alcohol en forma aguda sobre la morfometría trabecular y la resistencia a la tracción de la mandíbula en ratas adolescente Sprague Dawley (SD). El estudio estuvo compuesto por 24 ratas, de 7 semanas de edad, colocadas en el grupo control expuesto al alcohol [n=12 (6 machos y 6 hembras)] y alimentado en parejas [n=12 (6 machos y 6 hembras)]. El tratamiento de los grupos fue el siguiente; al grupo expuesto al alcohol y al control alimentado en parejas se les administró una dosis única diaria de 3 g/kg de alcohol al 20 % 3 días a la semana (días alternos) durante 7 días y una dosis equivalente calórica de maltosa dextrina mediante sonda oral, respectivamente. Los animales fueron sacrificados el día 7 mediante inyección de pentobarbital. Las mandíbulas se recolectaron y se escanearon utilizando un escáner 3D-microCT Nikon XTH 255L (Nikon Metrology, Lovaina, Bélgica), y las pruebas biomecánicas se realizaron utilizando una máquina de prueba de resistencia a la tracción universal Shimadzu (China). Después del escaneo y la reconstrucción, la morfometría trabecular se evaluó utilizando el software Volume Graphics Studio®. Se utilizó una prueba de flexión de 3 puntos para evaluar la resistencia a la tracción del hueso. Los hallazgos de nuestro estudio mostraron cambios en algunos parámetros trabeculares en el grupo de hembras expuestas al alcohol, mientras que los grupos de machos no se vieron afectados. No se observaron cambios en la resistencia a la tracción al comparar los grupos control de machos alimentados en parejas y los grupos expuestos al alcohol y al comparar los grupos control de las hembras alimentadas en parejas y los grupos expuestos al alcohol. Se observaron diferencias trabeculares y de resistencia a la tracción entre los sexos al comparar los grupos control de los machos alimentados en parejas y expuestos al alcohol con los grupos de control de hembras alimentadas en parejas y expuestas al alcohol. Estos hallazgos sugieren que el consumo excesivo de alcohol tiene efectos perjudiciales sobre la microarquitectura ósea en ratas hembras expuestas al alcohol y que se observan diferencias entre los sexos.
Assuntos
Animais , Masculino , Feminino , Ratos , Etanol/toxicidade , Consumo Excessivo de Bebidas Alcoólicas , Mandíbula/efeitos dos fármacos , Resistência à Tração , Fenômenos Biomecânicos , Densidade Óssea , Fatores Sexuais , Ratos Sprague-Dawley , Modelos Animais de Doenças , Concentração Alcoólica no Sangue , Osso Esponjoso/efeitos dos fármacosRESUMO
Mare endometrosis is a major reproductive problem associated with low fertility and is characterized by persistent inflammation, TGFß-1 signaling, and consequently, extracellular matrix deposition, which compromises endometrial glands. Mesenchymal stem cell-based products (MSCs), such as extracellular vesicles (EVs), have gained attention due to the regulatory effects exerted by their miRNA cargo. Here, we evaluated the impact of preconditioning equine adipose mesenchymal stem cells with TGFß-1 for short or long periods on the anti-fibrotic properties of secreted extracellular vesicles. MSCs were isolated from six healthy horses and exposed to TGFß-1 for 4, 24, and 0 h. The expression of anti-fibrotic and pro-fibrotic miRNAs and mRNAs in treated cells and miRNAs in the cargo of secreted extracellular vesicles was measured. The resulting EVs were added for 48 h to endometrial stromal cells previously induced to a fibrotic status. The expression of anti-fibrotic and pro-fibrotic genes and miRNAs was evaluated in said cells using qPCR and next-generation sequencing. Preconditioning MSCs with TGFß-1 for 4 h enriched the anti-fibrotic miRNAs (mir29c, mir145, and mir200) in cells and EVs. Conversely, preconditioning the cells for 24 h leads to a pro-fibrotic phenotype overexpressing mir192 and mir433. This finding might have implications for developing an EV-based protocol to treat endometrial fibrosis in mares.
Assuntos
Endométrio , Vesículas Extracelulares , Fibrose , Células-Tronco Mesenquimais , MicroRNAs , Fator de Crescimento Transformador beta1 , Animais , Cavalos , Feminino , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Vesículas Extracelulares/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/genética , Endométrio/metabolismo , Endométrio/citologia , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Células Estromais/metabolismo , Células Estromais/efeitos dos fármacos , Doenças dos Cavalos , Regulação da Expressão Gênica/efeitos dos fármacos , Endometriose/veterinária , Endometriose/metabolismo , Endometriose/genéticaRESUMO
In brief: FSH leads to glutamine dependence, which is required for mTORC1 activation and in consequence Sertoli cell proliferation. Abstract: The spermatogenic capacity of adult individuals depends on, among other factors, the number of Sertoli cells (SCs) that result from the proliferative waves during development. FSH upregulates SC proliferation at least partly, through the activation of the PI3K/Akt/mTORC1 pathway, among other mechanisms. It is widely known that mTORC1 is a sensor of amino acids. Among amino acids, glutamine acquires relevance since it might contribute to cell cycle progression through the modulation of mTORC1 activity. It has not been studied yet whether glutamine intervenes in FSH-mediated regulation of SC proliferation and cell cycle progression, or if FSH has any effect on glutamine metabolism. Eight-day-old rat SCs were incubated in culture media without glutamine or with glutamine in the absence or presence of a glutamine transporter inhibitor or a glutaminase activity inhibitor under basal conditions or stimulated with FSH. The results obtained show that FSH does not promote SC proliferation and mTORC1 activation in the absence of glutamine. Also, FSH modulates glutamine metabolism increasing glutaminase isoform 2 and reducing glutamine synthetaseexpression. FSH did not promote SC proliferation and mTORC1 activation when glutaminase activity was inhibited. The results suggest that glutamine or its metabolites might cooperate with FSH in the upregulation of SC proliferation through mTORC1. In addition, as FSH modulates glutamine metabolism through the induction of glutaminase isoform 2, the hormonal control of glutamine metabolism might be part of the intricate signaling network triggered by FSH, which is crucial to establish the population of mature SCs that supports the reproductive function.
Assuntos
Proliferação de Células , Hormônio Foliculoestimulante , Glutamina , Alvo Mecanístico do Complexo 1 de Rapamicina , Células de Sertoli , Animais , Glutamina/metabolismo , Glutamina/farmacologia , Masculino , Células de Sertoli/metabolismo , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/citologia , Hormônio Foliculoestimulante/farmacologia , Hormônio Foliculoestimulante/metabolismo , Proliferação de Células/efeitos dos fármacos , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Ratos , Células Cultivadas , Transdução de Sinais/efeitos dos fármacos , Glutaminase/metabolismo , Ratos Sprague-Dawley , Ratos WistarRESUMO
The present study investigated the effect of adding allopathic doxorubicin (DOX 0.3⯵g/mL), the vehicle of ultradiluted/dynamized doxorubicin (0.2â¯% ethanol), different dynamizations of ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH), both in the absence or presence of chemical stress induced by doxorubicin at 0.3⯵g/mL on follicular survival and activation, antioxidant capacity of the medium, Catalase activity (CAT), production of reactive protein thiol, maintenance of type I and III collagen fibers and accumulation of lipofuscin in porcine ovarian tissue cultured in vitro for 48â¯hours. To do this, part of the ovarian tissue fragments was fixed for the uncultured control and the rest were cultured in: MEM (cultured control), DOX 0.3⯵g/mL, Ethanol, DOX 6CH, DOX 12CH, DOX 30CH, DOX (0.3⯵g/mL) + DOX 6CH, DOX (0.3⯵g/mL) + DOX 12CH, DOX (0.3⯵g/mL) + DOX 30CH treatments. The results showed that, in general, ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH) mitigated the toxic effect of allopathic doxorubicin (0.3⯵g/mL) on the morphology of preantral follicles, the content of type I and III collagen fibers, and the production of lipofuscin in the tissue. However, only DOX (0.3⯵g/mL) + DOX 6CH attenuated the oxidative stress induced by DOX (0.3⯵g/mL), maintaining adequate CAT activity that was similar to the uncultured control. Additionally, when the three isolated ultradiluted/dynamized doxorubicin were considered, only DOX 12CH increased the reduced thiol levels compared to the uncultured control and MEM. In conclusion, supplementing the culture medium with ultradiluted/dynamized DOX (DOX 6CH, DOX 12CH and DOX 30CH) attenuated the toxicity induced by allopathic doxorubicin during the in vitro culture of pig preantral follicles enclosed in ovarian tissue.
Assuntos
Antibióticos Antineoplásicos , Doxorrubicina , Folículo Ovariano , Animais , Doxorrubicina/toxicidade , Feminino , Suínos , Antibióticos Antineoplásicos/toxicidade , Folículo Ovariano/efeitos dos fármacos , Catalase/metabolismo , Técnicas de Cultura de Tecidos , Lipofuscina/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Antioxidantes/farmacologia , Colágeno Tipo I/metabolismo , Ovário/efeitos dos fármacos , Compostos de Sulfidrila/metabolismo , Colágeno Tipo III/metabolismoRESUMO
In brief: The hypoglycemic drug metformin has shown reproductive effects in women, although its mechanism of action is not fully understood. In this study, we demonstrate the direct effects of metformin on the ovary of healthy mice, with no alterations in fertility. Abstract: Metformin is a hypoglycemic drug widely used in type-2 diabetes (T2D) patients. In recent years, this drug has been suggested as a treatment for gestational diabetes and recommended to women with ovarian hyperstimulation syndrome (PCOS) to increase the chances of pregnancy or avoid early miscarriages. However, the exact effects of metformin on the female reproductive tract in general, and on the ovary in particular, are still not completely understood. In this study, we analyzed the effect of metformin on fertility and ovarian physiology in healthy female mice. We found that this drug altered the estrous cycle, early follicular development, serum estradiol and progesterone levels, and ovarian steroidogenic enzyme expression. Moreover, ovarian angiogenesis was lower in metformin-treated animals compared with untreated ones, whereas natural or gonadotropin-induced fertilization rates remained unchanged. However, offspring of metformin-treated animals displayed decreased body weight at birth. In this work, we unraveled the main effects of metformin on the ovary, isolated from other conditions such as hyperglycemia and hyperandrogenism, which is essential for a better understanding of metformin's mechanisms of action on reproduction and fertility.
Assuntos
Ciclo Estral , Fertilidade , Hipoglicemiantes , Metformina , Ovário , Animais , Feminino , Metformina/farmacologia , Camundongos , Ovário/efeitos dos fármacos , Ovário/metabolismo , Fertilidade/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Ciclo Estral/efeitos dos fármacos , Gravidez , Estradiol/sangue , Progesterona/sangueRESUMO
(-)-Carvone, a ketone monoterpene, is the main component of essential oils from several medicinal plants and has been reported to have anti-arthriric, anticonvulsive, antidiabetic, anti-inflammatory, anticancer, and immunomodulatory effects. Therefore, this study aimed to investigate the spasmolytic activity of (-)-carvone in rodent models. The isolated virgin rat uterus was mounted in an organ bath apparatus, and the relaxing effect of ( -)-carvone and its mechanism of action were evaluated in tonic contractions induced by carbachol, KCl, PGF2α, or oxytocin. The animal model of primary dysmenorrhea was replicated with the injection of estradiol benzoate in female mice for three consecutive days, followed by intraperitoneal administration of oxytocin. Non-clinical acute toxicity evaluation was also performed. (-)-Carvone potency and effectiveness were larger in carbachol (pEC50 = 5.41 ± 0.14 and Emax = 92.63 ± 1.90% at 10-3 M) or oxytocin (pEC50 = 4.29 ± 0.17 and Emax = 86.69 ± 1.56% at 10-3 M) contractions. The effect of ( -)-carvone was altered in the presence of 4-aminopyridine, glibenclamide, L-NAME, or methylene blue. Mice pre-treated with (-)-carvone at a dose of 100 mg/kg showed a significant reduction in the number of writhing after oxytocin administration. No toxicity was observed after oral administration of 1 g/kg ( -)-carvone. Taken together, we showed that (-)-carvone reduced writhing by a spasmolytic effect, probably through the participation of KV and KATP channels and the nitric oxide pathway.
Assuntos
Monoterpenos Cicloexânicos , Monoterpenos , Ocitocina , Útero , Animais , Ocitocina/farmacologia , Feminino , Monoterpenos Cicloexânicos/farmacologia , Camundongos , Útero/efeitos dos fármacos , Monoterpenos/farmacologia , Contração Uterina/efeitos dos fármacos , Ratos , Ratos Wistar , Parassimpatolíticos/farmacologia , Relaxamento Muscular/efeitos dos fármacos , Carbacol/farmacologiaRESUMO
OBJECTIVES: Chemoprevention can be a treatment for potentially malignant lesions (PMLs). We aimed to evaluate whether artemisinin (ART) and cisplatin (CSP) are associated with apoptosis and immunogenic cell death (ICD) in vitro, using oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC) cell lines, and whether these compounds prevent OL progression in vivo. METHODS: Normal keratinocytes (HaCat), Dysplastic oral cells (DOK), and oral squamous cell carcinoma (SCC-180) cell lines were treated with ART, CSP, and ART + CSP to analyze cytotoxicity, genotoxicity, cell migration, and increased expression of proteins related to apoptosis and ICD. Additionally, 41 mice were induced with OL using 4NQO, treated with ART and CSP, and their tongues were histologically analyzed. RESULTS: In vitro, CSP and CSP + ART showed dose-dependent cytotoxicity and reduced SCC-180 migration. No treatment was genotoxic, and none induced expression of proteins related to apoptosis and ICD; CSP considerably reduced High-mobility group box-1 (HMGB-1) protein expression in SCC-180. In vivo, there was a delay in OL progression with ART and CSP treatment; however, by the 16th week, only CSP prevented progression to OSCC. CONCLUSION: Expression of proteins related to ICD and apoptosis did not increase with treatments, and CSP was shown to reduce immunogenic pathways in SCC-180, while reducing cell migration. ART did not prevent the malignant progression of OL in vivo; CSP did despite significant adverse effects.
Assuntos
Apoptose , Artemisininas , Movimento Celular , Cisplatino , Progressão da Doença , Leucoplasia Oral , Neoplasias Bucais , Artemisininas/farmacologia , Animais , Leucoplasia Oral/patologia , Leucoplasia Oral/tratamento farmacológico , Humanos , Cisplatino/farmacologia , Camundongos , Neoplasias Bucais/patologia , Neoplasias Bucais/tratamento farmacológico , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proteína HMGB1/metabolismo , Antineoplásicos/farmacologiaRESUMO
This study investigates the toxic effects of the insecticide spinetoram on the model organism Bombyx mori (Linnaeus) and explores the potential ameliorative properties of O-Vanillin. Sub-lethal concentrations of spinetoram were given to silkworm larvae via oral feed, resulting in reduced body weight, larval length, and impaired cocoon characteristics. A study of the enzymatic and non-enzymatic antioxidants revealed oxidative stress in the gut, fat body, and silk gland tissues, characterized by decreased antioxidants and increased lipid peroxidation. However, post-treatment with O-Vanillin effectively mitigated these toxic effects, preserving antioxidant capacities and preventing lipid peroxidation. Additionally, O-Vanillin prevented the loss of body weight and improved cocoon characteristics. At the histological level, spinetoram exposure caused mild histological damage in the gut, fat body, and silk gland. However, O-Vanillin post-treatment had ameliorative effects and mitigated the histological damages. To delve deeper into the mechanism of amelioration of O-Vanillin, in silico studies were used to study the interaction between an important xenobiotic metabolism protein of the Bombyx mori, i.e., Cytochrome p450, specifically CYP9A19, and O-Vanillin. We performed blind molecular docking followed by molecular dynamic simulation, and the results demonstrated stable binding interactions between O-Vanillin and CYP9A19, a cytochrome P450 protein in silkworm, belonging to the subfamily CYP9A, suggesting a potential role for O-vanillin in modulating xenobiotic metabolism.
Assuntos
Benzaldeídos , Bombyx , Inseticidas , Larva , Estresse Oxidativo , Animais , Bombyx/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Benzaldeídos/farmacologia , Larva/efeitos dos fármacos , Simulação de Acoplamento Molecular , Antioxidantes , Peroxidação de Lipídeos/efeitos dos fármacosRESUMO
Coffee berry borer (CBB) Hypothenemus hampei is a major biotic threat to coffee production worldwide. Studies have reported negative effects on CBB by oil-based formulations of neem (Azadirachta indica), but little information is available for other neem-extract formulations. This study evaluated CBB preference and performance in arabica coffee fruits and artificial diet treated with a neem-extract formulation (Openeem Plus®) in the field and laboratory conditions. Field experiments were performed using CBB females artificially infested in cherry or green coffee fruits confined in voile-fabric cages tied to branches of neem-treated and control plants, recording the adult mortality and offspring production. Dual-choice and no-choice bioassays assessed CBB preference and development in fruits and artificial diet treated with the neem extract compared to controls in the laboratory, respectively. As main results obtained in the field and laboratory experiments, the neem extract significantly reduced CBB oviposition in both cherry and green fruits, as well as in artificial diet compared to controls. However, the botanical product did not affect CBB adult survival and preference in the laboratory bioassays. The neem extract is promising for use in pest management strategies in sustainable arabica coffee crops by reducing CBB oviposition and offspring. These effects can contribute to lowering the pest population buildup along the crop cycle and damage potential to coffee production.
Assuntos
Azadirachta , Coffea , Frutas , Oviposição , Gorgulhos , Animais , Feminino , Oviposição/efeitos dos fármacos , Dieta , Controle de Insetos/métodosRESUMO
Novel open-chain merocytochalasans, perochalasins A-C (1-3), containing an unusual N-O six-membered heterocyclic moiety, were isolated from cultures of the marine-derived Peroneutypa sp. M16 fungus, along with cytochalasin Z27 (4), cytochalasin Z28 (5), [12]-cytochalasin (6), and phenochalasin B (7). The structures of compounds 1-3 were established by analysis of the spectroscopic data. Full genome sequencing of Peroneutypa sp. M16 enabled the identification of a cytochalasan biosynthetic gene cluster and a proposal for the biosynthetic assembly of perochalasins. The proposal is supported by the nonenzymatic conversion of phenochalasin B (7) into 1-3, based on isotope-labeled hydroxylamine (15NH2OH and ND2OD) feeding studies in vivo and in vitro. In contrast to other merocytochalasans, these are the first cytochalasans confirmed to arise via nucleophilic addition and at a distinct location from the reactive macrocycle olefin, potentially expanding further the range of merocytochalasans to be discovered or engineered. Cytochalasin Z27 (4) exhibited antiplasmodial activities in the low micromolar range against the chloroquine-sensitive Plasmodium falciparum 3D7 strain as well as against resistant strains of the parasite (Dd2, TM90C6B, and 3D7r_MMV848).
Assuntos
Citocalasinas , Citocalasinas/farmacologia , Citocalasinas/química , Citocalasinas/biossíntese , Citocalasinas/isolamento & purificação , Estrutura Molecular , Plasmodium falciparum/efeitos dos fármacos , Antimaláricos/farmacologia , Antimaláricos/química , Família MultigênicaRESUMO
OBJECTIVE: Tumor hypoxia is associated with a poorer prognosis in cancer patients and can diminish the efficacy of radiation therapy (RT). This study investigates the potential of metformin to enhance radiosensitivity in hypoxic cancer cells. METHODS: Preliminary experiments were conducted to validate the impact of hypoxia on radiation response. Reactive oxygen species (ROS) levels, cell migration, and cell death were assessed in hypoxic, radiated cells treated with metformin. Proteomic and ontological analyses were employed to identify molecular targets associated with the radiosensitizing effect of metformin. Proteomic and ontological findings were validated through patient samples and in vitro studies. RESULTS: Metformin amplified cell death, induced DNA fragmentation, decreased cell migration, and elevated ROS levels in hypoxic, radiated cells. Proteomic analyses revealed that GAPDH and TAGLN2 were identified as pivotal targets linked to the radiosensitizing effect of metformin. Oral cancer patients exhibited elevated levels of TAGLN2 and reduced levels of GAPDH. Metformin downregulated TAGLN2 and upregulated GAPDH in hypoxic, radiated cells. Additionally, metformin reduced levels of mutated p53. CONCLUSIONS: This study suggests that metformin can enhance radiosensitivity in hypoxic cells, operating through modulation of GAPDH and TAGLN2. Furthermore, metformin effectively reduces mutated p53 levels in radiated cells under hypoxic conditions.
Assuntos
Carcinoma de Células Escamosas , Metformina , Neoplasias Bucais , Radiossensibilizantes , Humanos , Metformina/farmacologia , Metformina/uso terapêutico , Neoplasias Bucais/radioterapia , Radiossensibilizantes/farmacologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Tolerância a Radiação/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Proteômica , Gliceraldeído-3-Fosfato Desidrogenases , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora) , Hipóxia Celular/efeitos dos fármacos , Hipóxia Tumoral/efeitos dos fármacosRESUMO
This study assessed the physicochemical and antibiofilm properties of white mineral trioxide aggregate (MTA) associated with 1 or 2% of farnesol. Setting time was evaluated based on ISO 6876/2012. Radiopacity was evaluated by radiographic analysis. pH was assessed after time intervals of 1, 3, 7, 14, 21, and 28 days. Solubility (% mass loss) and volumetric change (by micro-CT) of the cements were evaluated after immersion in distilled water. The presence of voids inside the materials was assessed by using micro-CT. Antibiofilm activity against Enterococcus faecalis was evaluated by crystal violet assay and the modified direct contact test performed with biofilm previously formed on bovine root dentin for 14 days. Data were submitted to ANOVA/Tukey tests with 5% significance level. The incorporation of farnesol into MTA increased its setting time, but decreased its solubility at 30 days and its volumetric loss in all periods (p < 0.05). Radiopacity and solubility after 7 days were similar among the materials (p > 0.05). The association of farnesol showed the highest pH value after 1 and 3 days (p < 0.05). The association of farnesol with MTA promoted a decrease in the presence of voids, and increased the antimicrobial activity on biofilm biomass of E. faecalis (p < 0.05). In conclusion, the addition of farnesol can be suggested to improve the antimicrobial properties and the consistency of MTA.
Assuntos
Compostos de Alumínio , Biofilmes , Compostos de Cálcio , Combinação de Medicamentos , Enterococcus faecalis , Farneseno Álcool , Teste de Materiais , Óxidos , Materiais Restauradores do Canal Radicular , Silicatos , Solubilidade , Silicatos/farmacologia , Silicatos/química , Óxidos/farmacologia , Óxidos/química , Biofilmes/efeitos dos fármacos , Compostos de Cálcio/farmacologia , Compostos de Cálcio/química , Enterococcus faecalis/efeitos dos fármacos , Compostos de Alumínio/farmacologia , Compostos de Alumínio/química , Farneseno Álcool/farmacologia , Farneseno Álcool/química , Concentração de Íons de Hidrogênio , Fatores de Tempo , Bovinos , Materiais Restauradores do Canal Radicular/farmacologia , Materiais Restauradores do Canal Radicular/química , Animais , Análise de Variância , Reprodutibilidade dos Testes , Dentina/efeitos dos fármacos , Valores de Referência , Propriedades de Superfície/efeitos dos fármacosRESUMO
This is a nonclinical, controlled, and triple-blind study to investigate the effects of codeine-associated geraniol on the modulation of orofacial nociception and its potential central nervous system depressing effect in an animal model. The orofacial antinociceptive activity of geraniol in combination with codeine was assessed through the following tests: (i) formalin-induced pain, (ii) glutamate-induced pain, and (iii) capsaicin-induced pain. Six animals were equally distributed into six groups and received the following treatments, given intraperitoneally (i.p.) 30 minutes before the experiments: a) geraniol/codeine 50/30 mg/kg; b) geraniol/codeine 50/15 mg/kg; c) geraniol/codeine 50/7.5 mg/kg; d) geraniol 50 mg/kg; e) codeine 30 mg/kg (positive control); or f) 0.9% sodium chloride (negative control). We performed pain behavior analysis after the injection of formalin (20 µL, 20%), glutamate (20 µL, 25 µM), and capsaicin (20 µL, 2.5 µg) into the paranasal region. Rubbing time of the paranasal region by the hind or front paw was used as a parameter. In the neurogenic phase of the formalin test, the geraniol/codeine at 50/7.5 mg/kg was able to promote the maximum antinociceptive effect, reducing nociception by 71.9% (p < 0.0001). In the inflammatory phase of the formalin test, geraniol/codeine at 50/30 mg/kg significantly reduced orofacial nociception (p < 0.005). In the glutamate test, geraniol/codeine at 50/30 mg/kg reduced the rubbing time by 54.2% and reduced nociception in the capsaicin test by 66.7% (p < 0.005). Geraniol alone or in combination does not promote nonspecific depressing effects on the central nervous system. Based on our findings, we suggest the possible synergy between geraniol and codeine in the modulation of orofacial pain.
Assuntos
Monoterpenos Acíclicos , Analgésicos , Capsaicina , Codeína , Dor Facial , Medição da Dor , Terpenos , Animais , Codeína/farmacologia , Dor Facial/induzido quimicamente , Dor Facial/tratamento farmacológico , Monoterpenos Acíclicos/farmacologia , Masculino , Medição da Dor/efeitos dos fármacos , Capsaicina/farmacologia , Terpenos/farmacologia , Analgésicos/farmacologia , Camundongos , Fatores de Tempo , Modelos Animais de Doenças , Reprodutibilidade dos Testes , Formaldeído , Ácido Glutâmico , Resultado do Tratamento , Nociceptividade/efeitos dos fármacos , Análise de Variância , Estatísticas não Paramétricas , Comportamento Animal/efeitos dos fármacosRESUMO
This study aimed to assess the influence of streptozotocin (STZ)-induced diabetes on the nociceptive behavior evoked by the injection of hypertonic saline (HS) into the masseter muscle of rats. Forty male rats were equally divided into four groups: a) isotonic saline control, which received 0.9% isotonic saline (IS), (Ctrl-IS); b) hypertonic saline control, which received 5% HS (Ctrl-HS); c) STZ-induced diabetic, which received IS, (STZ-IS); d) STZ-induced diabetic, which received HS (STZ-HS). Experimental diabetes was induced by a single intraperitoneal injection of STZ at dose of 60 mg/kg dissolved in 0.1 M citrate buffer, and 100 µL of HS or IS were injected into the left masseter to measure the nociceptive behavior. Later on, muscle RNA was extracted to measure the relative expression of the following cytokines: cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF-α), and interleukins (IL)-1ß, -2, -6, and -10. One-way analysis of variance (ANOVA) was applied to the data (p < 0.050). We observed a main effect of group on the nociceptive response (ANOVA: F = 11.60, p < 0.001), where the Ctrl-HS group presented the highest response (p < 0.001). However, nociceptive response was similar among the Ctrl-IS, STZ-IS, and STZ-HS group (p > 0.050). In addition, the highest relative gene expression of TNF-α and IL-6 was found in the masseter of control rats following experimental muscle pain (p < 0.050). In conclusion, the loss of somatosensory function can be observed in deep orofacial tissues of STZ-induced diabetic rats.
Assuntos
Citocinas , Diabetes Mellitus Experimental , Músculo Masseter , Ratos Wistar , Estreptozocina , Animais , Masculino , Músculo Masseter/efeitos dos fármacos , Músculo Masseter/fisiopatologia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/fisiopatologia , Análise de Variância , Citocinas/análise , Solução Salina Hipertônica/farmacologia , Medição da Dor , Fatores de Tempo , Reprodutibilidade dos Testes , Dor Facial/fisiopatologia , Distribuição Aleatória , RatosRESUMO
Insect growth regulators (IGRs) have been playing a major role in the effective management of a range of stored product insect pests including species that have developed resistance to major groups of insecticides, such as organophosphates (OPs) and synthetic pyrethroids (SPs). In the present study, we evaluated the efficacy of S-methoprene alone and in combination with piperonyl butoxide (PBO), an adjuvant component of insecticides for synergy, against two strains, Lab-S (susceptible) and Met-R (Methoprene resistant) of an economically important pest species, the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). Adults of both Lab-S and Met-R strains were exposed to wheat treated with multiple concentrations of S-methoprene ranging from 0.001 to 0.01 and 10 to 60 mg/kg, respectively, alone and in combination with PBO. A variety of concentrations, including 0.27, 0.53, 0.80, and 1.07 g/kg, were evaluated for PBO. Mortality of adults and percent reduction in progeny were assessed after 14 and 65 days of treatment, respectively. As anticipated, the adult mortality rates of both strains were not significantly affected by S-methoprene alone. However, the number of progeny produced confirmed that the Met-R strain exhibited a high level of resistance to S-methoprene alone, with over 50 F1 progeny adults surviving in wheat treated with the maximal rate, 10 mg/kg. In contrast, the toxicity of S-methoprene was increased against the same resistant strain (Met-R), by 0.80 or 1.07 g/kg of PBO in combination treatment, resulting in a significant reduction in progeny numbers (25 adults per container). Although the tested concentrations of S-methoprene and PBO were well above the currently registered rate globally, our results highlight the fact that PBO enhances the toxicity of S-methoprene to some extent, reaffirming that the mode of action of the latter involves the inhibition of mixed-function oxidases (MFOs) and highlights the need for further research into developing potential binary or triplet formulations containing these two active ingredients (AIs).