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SUMMARY: Stroke is one of the main causes of death and disability worldwide. The great impact on the quality of life of the population and on the health system justifies that we seek relevant alternatives to reduce the incidence and improve the treatment and recovery of patients affected by this disease. Physical exercise appears as an important tool in this scenario, being already pointed out as a possible therapeutic approach for the prevention of non-contagious chronic diseases. In this context, biomarkers such as miRNAs that respond to physical exercise and are directly related to several epigenetic mechanisms appear. Therefore, explaining the molecular mechanisms involved during physical exercise will lead to a better understanding of each stimulus and the dose to be used to better respond to each situation, thus being a promising approach for the evolution of prescription and control of training and processes recovery from various diseases, including stroke. Forty-eight Wistar rats were used, divided into four experimental groups: control group, ischemia group, physical exercise group and exercise + ischemia group. Real-time PCR methodology was used to analyze the expression of miRNAs: miR-126, miR-133b and miR-221. In our study we observed a significant difference in the expression of miR- 221 between the control group and the others groups. However, microRNAs: miR-126 and miR-133b do not show significant differences in expression between groups.

RESUMEN: El ictus es una de las principales causas de muerte y discapacidad en todo el mundo. El gran impacto en la calidad de vida de la población y en el sistema de salud justifica buscar alternativas pertinentes para reducir la incidencia y mejorar el tratamiento y recuperación de los pacientes afectados por esta enfermedad. El ejercicio físico aparece como una herramienta importante en este escenario, siendo ya señalado como un posible abordaje terapéutico para la prevención de enfermedades crónicas no contagiosas. En este contexto, aparecen biomarcadores como los miRNAs que responden al ejercicio físico y están directamente relacionados con varios mecanismos epigenéticos. Por lo tanto, explicar los mecanismos moleculares involucrados durante el ejercicio físico conducirá a una mejor comprensión de cada estímulo y la dosis a utilizar para responder mejor a cada situación, siendo así un enfoque prometedor para la evolución de la prescripción, el control del entrenamiento y los procesos de recuperación de diversas enfermedades, incluido el accidente cerebrovascular. Se utilizaron cuarenta y ocho ratas Wistar, divididas en cuatro grupos experimentales: grupo control, grupo isquemia, grupo ejercicio físico y grupo ejercicio + isquemia. Se utilizó la metodología de PCR en tiempo real para analizar la expresión de miRNAs: miR-126, miR-133b y miR-221. En nuestro estudio observamos una diferencia significativa en la expresión de miR-221 entre el grupo control y los demás grupos. Sin embargo, los microARN: miR-126 y miR-133b no mostraron diferencias significativas en la expresión entre grupos.

MicroRNA-27b Expression in Rats Submitted to Physical Exercise and Experimental Cerebral Ischemia
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De-Russi, Brenno Marco

; Porsani, Lucas Barbosa

; Cirino, Mucio Luiz de Assis

; Silva, Jairo Pinheiro da

; Turra, Letícia Passi

; Novais, Paulo Cezar

; Lizarte-Neto, Fermino Sanches

; Fazan, Valéria de Paula Sassoli

; Thomazini, José Antônio

; Colli, Benedicto Oscar

; Tirapelli, Daniela Pretti da Cunha

; Tirapelli, Luis Fernando

; Carvalho, Camila Albuquerque Melo de

International Journal of Morphology

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Jun 2021, Volume 39 Nº 3 Páginas 754 - 758

Resumo: En Es | Texto: En Es | PDF: En | PDF: Es

SUMMARY: Cerebral ischemia has not only a high mortality rate, which is the second leading cause of death worldwide, but is also responsible for severe disabilities in working age individuals, generating enormous public expending for treatment and rehabilitation of the affected individuals. The role of microRNAs in the pathophysiology of cerebral ischemia has been highlighted in current investigations. In addition, recent studies have also highlighted physical exercise as a possible protective factor both in the prevention and in the effects of cerebral ischemia, placing it as an important study resource. Thus, we investigated the role of physical exercise in experimental cerebral ischemia associated with the expression of microRNA-27b. 16 animals were used, divided into four experimental groups: Control, Physical Exercise, Cerebral Ischemia and Cerebral Ischemia associated with Physical Exercise. The real-time PCR methodology was used to analyze the expression of microRNA-27b. Although there were no statistically significant differences in the expression of microRNA-27b between the groups studied, the increased expression of microRNA-27b in the Physical Exercise group indicates its neuroprotective role in the pathophysiology of cerebral ischemia.

RESUMEN: La isquemia cerebral no solo tiene una alta tasa de mortalidad y es la segunda causa principal de muerte en todo el mundo, sino también es la causa de enfermedades invalidantes en personas en edad laboral, lo que genera un gasto público enorme para el tratamiento y la rehabilitación de las personas afectadas. El papel de los microARN en la fisiopatología de la isquemia cerebral se ha destacado en las investigaciones actuales. Además, estudios recientes también han destacado el ejercicio físico como un posible factor protector tanto en la prevención como en los efectos de la isquemia cerebral, situándolo como un importante recurso de estudio. Por lo tanto, investigamos el papel del ejercicio físico en la isquemia cerebral experimental asociada con la expresión del microARN-27b. Se utilizaron 16 animales, divididos en cuatro grupos experimentales: Control, Ejercicio Físico, Isquemia Cerebral e Isquemia Cerebral asociada al Ejercicio Físico. Se utili- zó la metodología de PCR en tiempo real para analizar la expresión de microARN-27b. Aunque no se observaron diferencias estadísticamente significativas en la expresión de microARN-27b entre los grupos estudiados, la mayor expresión de microARN-27b en el grupo de Ejercicio Físico indica su papel neuroprotector en la fisiopatología de la isquemia cerebral.

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Expression of AIF, PARP, and miRNAs MIR-145, MIR-210, and MIR-486 Associated with Apoptosis in the Corpus Cavernosum Of Rats Subjected to Chronic Alcoholism Model
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Sarraipo, Vagner Schiavoni

; Lizarte Neto, Fermino Sanches

; Carvalho, Camila Albuquerque Mello de

; Silva, Jairo Pinheiro da

; Durand, Marina Toledo

; Schimming, Bruno Cezar

; Martinez, Marcelo

; Galli, Maria de Fátima Tazima Sorita

; Buono, Fabianna de Oliveira

; Tiezzi, Maria Fernanda Barbirato da Mata

; Novais, Paulo Cezar

; Gula, Isabella Stracieri

; Thomazini, José Antônio

; Fazan, Valeria Paula Sassoli

; Rodrigues, Andressa Romualdo

; Tirapelli, Daniela Pretti da Cunha

; Cirino, Mucio Luiz de Assis

; Molina, Carlos Augusto Fernandes

; Junior, Silvio Tucci

; Tirapelli, Luis Fernando

International Journal of Morphology

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Dez 2020, Volume 38 Nº 6 Páginas 1639 - 1644

Resumo: En Es | Texto: En Es | PDF: En | PDF: Es

SUMMARY: Previous studies from our group described the consequences of using ethanol on penile erection. Nevertheless, the molecular mechanisms surrounding microRNAs, apoptosis process and their relationship with erectile dysfunction associated with alcohol consumption are still poorly understood. The objective of this analysis was to evaluate the mechanism of apoptosis by the expression of AIF and PARP, as well as their regulatory microRNAs: miR-145, miR-210 and miR-486, in the corpus cavernosum of rats submitted to a semivoluntary alcoholism model. For this study 24 Wistar rats were divided into two groups: control (C) and treated with 20 % ethanol (A) for seven weeks. The corpus cavernosum samples were prepared for immunohistochemical analysis of AIF and PARP protein expression, and microRNAs miR-145, miR-210, miR-486 gene expression in cavernous tissue was performed by real time PCR. The immunohistochemical analysis showed little nuclear positive labeling for the protein PARP and AIF in the corpus cavernosum of control and ethanol treated animals. After analysis of miR-145, -210 and -486 microRNA expression in the 12 animals studied, no results were found with significant statistical difference between the control and alcoholized groups. The expression of AIF and PARP and their regulatory microRNAs involved in apoptotic process (miR-145, miR-210 and miR-486) were not altered in the corpus cavernosum of rats submitted to semivoluntary alcoholism.

RESUMEN: Estudios previos de nuestro grupo describieron las consecuencias del uso de etanol en la erección del pene. Sin embargo, los mecanismos moleculares que rodean a los microARN, el proceso de apoptosis y su relación con la disfunción eréctil asociada con el consumo de alcohol aún no se conocen bien. El objetivo de este análisis fue evaluar el mecanismo de apoptosis mediante la expresión de AIF y PARP, así como sus microARN reguladores: miR-145, miR-210 y miR-486, en el cuerpo cavernoso de ratas sometidas a un modelo de alcoholismo semivoluntario. Se dividieron 24 ratas Wistar en dos grupos: control (C) grupo de ratas tratadas con etanol al 20 % (A) durante siete semanas. Las muestras del cuerpo cavernoso se prepararon para el análisis inmunohistoquímico de la expresión de la proteína AIF y PARP, y la expresión del gen microRNAs miR-145, miR-210, miR-486 en tejido cavernoso se realizó por PCR en tiempo real. El análisis inmunohistoquímico mostró escaso etiquetado nuclear positivo para la proteína PARP y AIF en el cuerpo cavernoso de los animales de control y tratados con etanol. Después del análisis de la expresión de microARN miR-145, -210 y -486 no se encontraron resultados con diferencias estadísticas significativas entre los grupos control y alcoholizados. La expresión de AIF y PARP y sus microARN reguladores involucrados en el proceso apoptótico (miR-145, miR-210 y miR-486) no se alteraron en el cuerpo cavernoso de las ratas sometidas a alcoholismo semivoluntario.

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Morphometric Analysis and Pattern of Protein and Gene Expression of Apoptosis in Focal Cerebral Ischemia in Rats and the Neuroprotetive Action of Hypothermia and Ketoprofen
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Tirapelli, Daniela P. C

; Cirino, Mucio Luiz de Assis

; Carvalho, Camila A. Melo de

; Novais, Paulo Cezar

; Figueredo, Rafael Zimak

; Porsani, Lucas Barbosa

; Gula, Isabella Stracieri

; Rodrigues, Andressa Romualdo

; Silva, Jairo Pinheiro da

; Pereira, Adriana Lis

; Lizarte Neto, Fermino Sanches

; Schimming, Bruno Cesar

; Tazima, Maria de Fátima Galli Sorita

; Fazan, Valéria de Paula Sassoli

; Carlotti-Jr, Carlos Gilberto

; Tirapelli, Luis Fernando

; Colli, Benedicto Oscar

International Journal of Morphology

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Jun 2020, Volume 38 Nº 3 Páginas 523 - 529

Resumo: En Es | Texto: En Es | PDF: En | PDF: Es

SUMMARY: This study aimed to investigate the morphometric and the pattern of protein and gene expression related to the extrinsic apoptotic pathway in experimental focal cerebral ischemia and the hole of neuroprotection with hypothermia and ketoprofen. For this analysis, 120 rats were randomly divided into 3 groups (20 animals each): control - no surgery (20 animals); sham - simulation of surgery (20 animals); ischemic - focal ischemia for 1 hour, without reperfusion (80 animals) and divided into four subgroups with 20 animals each: ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the apoptosis genes (Fas, c-Flip, caspase-8 and caspase-3) and the apoptosis protein caspase-3 were evaluated by quantitative real-time PCR and immunohistochemistry, respectively. Hypo expression of genes of extrinsic pathway of apoptosis was observed: Fas receptor, c-Flip and caspase-8 in the ischemics areas. Increases in the gene and protein caspase-3 in the ischemic areas were also observed, and these increases were reduced by hypothermia and ketoprofen, also noted in the morphometric study. The caspases-3 increase suggests that this gene plays an important role in apoptosis, probably culminating in cell death and that the neuroprotective effect of hypothermia and ketoprofen is involved.

RESUMEN: Este estudio tuvo como objetivo investigar la morfometría y el patrón de expresión de proteínas y genes relacionados con la vía apoptótica extrínseca en la isquemia cerebral focal experimental y el agujero de neuroprotección con hipotermia y ketoprofeno. Se dividieron aleatoriamente 120 ratas en 3 grupos (20 animales cada uno): control - sin cirugía (20 animales); simulación - simulación de cirugía (20 animales); isquemia isquemia focal durante 1 hora, sin reperfusión (80 animales) y dividida en cuatro subgrupos con 20 animales cada uno: isquemia + hipotermia intraisquémica; isquemia + ketoprofeno intravenoso previo, e isquemia + hipotermia y ketoprofeno. El volumen del infarto se midió utilizando un análisis morfométrico de áreas de infarto definidas por cloruro de trifenil tetrazolio y los patrones de expresión de los genes de apoptosis (Fas, c-Flip, caspase-8 y caspase-3) y la proteína de apoptosis caspase-3 fueron evaluados por PCR cuantitativa en tiempo real e inmunohistoquímica, respectivamente. Se observó hipoexpresión de genes de la vía extrínseca de la apoptosis: receptor Fas, c-Flip y caspasa-8 en las áreas isquémicas. También se observaron aumentos en el gen y la proteína caspasa-3 en las áreas isquémicas y estos aumentos se redujeron por hipotermia y ketoprofeno, también observado por estudio morfométrico. El aumento de caspasas-3 sugiere que este gen tiene un papel importante en la apoptosis, y probable causa de muerte celular, involucrando el efecto neuroprotector de la hipotermia y el ketoprofeno.

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AIF, PARP-1 and MicroRNA-9 Expression in Cerebral Ischemia Associated to Alcoholism Experimental Model
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Abrahão, Dayana Pousa Siqueira

; Lizarte Neto, Fermino Sanches

; Rodrigues, Andressa Romualdo

; Cirino, Múcio Luiz de Assis

; Silva, Jairo Pinheiro da

; Sarraipo, Vagner Schiavoni

; Carvalho, Camila Albuquerque Melo de

; Tazima, Maria de Fátima Galli Sorita

; Carlotti-Jr., Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

; Tirapelli, Daniela Pretti da Cunha

International Journal of Morphology

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Jun 2020, Volume 38 Nº 3 Páginas 616 - 621

Resumo: En Es | Texto: En Es | PDF: En | PDF: Es

SUMMARY: The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.

RESUMEN: El consumo crónico de alcohol provoca un empeoramiento de los eventos que siguen a la isquemia cerebral. Estos eventos están regulados a través de la expresión de varios genes y microRNA. El objetivo de este trabajo fue analizar y describir el perfil de expresión de las proteínas PARP y AIF y microRNA-9 en ratas sometidas a isquemia cerebral focal, asociadas o no, con el modelo de alcoholismo crónico. Veinte ratas Wistar adultas se dividieron en: grupo control, isquémico alcohólico, e isquémico / alcoholizado para análisis inmunohistoquímico y expresión de genes microRNA-9. Resultados: Hubo una reducción en la expresión de proteínas de PARP-1 y un marcado positivo para AIF en el grupo isquémico / alcoholizado. No se observó una expresión significativa en el microRNA-9. La asociación de la isquemia con el consumo crónico de alcohol promovió una tendencia a la baja expresión de microRNA-9, baja expresión de PARP1 y alta expresión de AIF, lo que indica una interferencia en el efecto protector de microRNA-9 en los otros grupos.

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Chronic alcoholism associated with diabetes induced apoptosis in the corpus cavernosum of rats
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Schimming, Bruno Cesar

; Cirino, Mucio Luiz de Assis

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cezar

; Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Molina, Carlos Augusto Fernandes

; Tirapelli, Luis Fernando

; Tucci Junior, Silvio

Acta Cirúrgica Brasileira

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2020, Volume 35 Nº 8 elocation e202000805

Resumo: En | Texto: En | PDF: En

Abstract Purpose To evaluate the effects of alcohol exposure and diabetes on apoptotic process in the corpus cavernosum. Methods Forty eight male Wistar rats were divided into four groups: control, diabetic, alcoholic and diabetic-alcoholic. Samples of the corpus cavernosum were prepared to study protein expression of apoptotic genes (Caspases-3 and 9) by immunohistochemistry and Real-Time PCR. Results The immunoreactivity of Caspases-3 and -9 was diffuse and higher in the treated groups though there was no significant difference between the experimental groups, only when compared with the control group. An increase was observed in the gene expression of Caspases-9 in the diabetic and ethanol-diabetic groups when compared with control and ethanol groups. Conclusions The association of these factors (ethanol and diabetes) probably can affect the apoptosis mechanism in lesions of the cavernous tissue in the rat penis. Both gene and protein expression of Caspase-9 in diabetic and ethanol-diabetic groups suggest the involvement of the apoptosis cascade from this study model.

https://doi.org/10.1590/s0102-865020200080000005 723 downloads

Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model
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Meirelles, Rogério José de Azevedo

; Lizarte Neto, Fermino Sanches

; Cirino, Mucio Luiz de Assis

; Novais, Paulo Cezar

; Gula, Isabella Stracieri

; Silva, Jairo Pinheiro da

; Tazzima, Maria de Fátima Galli Sorita

; Fazan, Valéria Paula Sassoli

; Durand, Marina Toledo

; Tirapelli, Daniela Pretti da Cunha

; Carvalho, Camila Albuquerque Melo de

; Schimming, Bruno César

; Molina, Carlos Augusto Fernandes

; Tucci Junior, Silvio

; Tirapelli, Luis Fernando

Acta Cirúrgica Brasileira

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2020, Volume 35 Nº 3 elocation e202000305

Resumo: En | Texto: En | PDF: En

Abstract Purpose To evaluate the effect of chronic alcoholism on morphometry and apoptosis mechanism and correlate with miRNA-21 expression in the corpus cavernosum of rats. Methods Twenty-four rats were divided into two experimental groups: Control (C) and Alcoholic group (A). After two weeks of an adaptive phase, rats from group A received only ethanol solution (20%) during 7 weeks. The morphometric and caspase-3 immunohistochemistry analysis were performed in the corpus cavernosum. The miRNA-21 expression was analyzed in blood and cavernous tissue. Results Chronic ethanol consumption decreased cavernosal smooth muscle area of alcoholic rats. The protein expression of caspase 3 in the corpus cavernosum was higher in A compared to the C group. There was no difference in the expression of miRNA-21 in serum and cavernous tissue between the groups. Conclusion Chronic ethanol consumption reduced smooth muscle area and increased caspase 3 in the corpus cavernosum of rats, without altered serum and cavernosal miR-21 gene expression.

https://doi.org/10.1590/s0102-865020200030000005 715 downloads

Analysis of Caspase-9 protein and microRNAs miR-21, miR-126 and miR-155 related to the apoptosis mechanism in the cerebellum of rats submitted to focal cerebral ischemia associated with an alcoholism model
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SILVA, Jairo Pinheiro da

; LIZARTE NETO, Fermino Sanches

; CIRINO, Mucio Luiz de Assis

; CARVALHO, Camila Albuquerque Melo de

; CARLOTTI JR, Carlos Gilberto

; COLLI, Benedicto Oscar

; TIRAPELLI, Daniela Pretti da Cunha

; TIRAPELLI, Luís Fernando

Arquivos de Neuro-Psiquiatria

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Out 2019, Volume 77 Nº 10 Páginas 689 - 695

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO O objetivo deste estudo foi analisar o cerebelo de ratos submetidos à isquemia cerebral focal experimental, por oclusão da artéria cerebral média por 90 minutos, seguida de reperfusão por 48 horas, associada a um modelo de alcoolismo. Métodos Foram utilizados 50 ratos Wistar adultos, subdivididos em cinco grupos experimentais: grupo controle (C): animais submetidos apenas à anestesia; grupo sham (S): animais submetidos à simulação completa do procedimento cirúrgico; grupo isquêmico (I): animais submetidos à isquemia cerebral focal por 90 minutos, seguidos de reperfusão por 48 horas; grupo alcoólico (A): animais que receberam etanol absoluto diário diluído em 20% em água por quatro semanas; e grupo isquêmico e alcoólico (I + A): animais que recebem o mesmo tratamento do grupo A e, após quatro semanas, submetidos à isquemia cerebral focal por 90 minutos, seguidos de reperfusão por 48 horas. As amostras de cerebelo foram coletadas e a análise imuno-histoquímica da proteína Caspase-9 e a análise sérica por RT-PCR dos microRNAs miR-21, miR-126 e miR155 foram realizadas. Resultados A expressão de Caspase-9 foi maior nos grupos I, A e I + A. Nas análises de microRNAs, o miR-126 foi maior nos grupos A e I + A, o miR-155 foi maior nos grupos I e I + A. Conclusões Concluímos que a apoptose ocorre no córtex cerebelar, mesmo distante do foco isquêmico, e que os microRNAs 126 e 155 mostram uma correlação com a apoptose celular em ratos isquêmicos e submetidos ao modelo crônico de álcool.

ABSTRACT This study aimed to analyze the cerebellum of rats submitted to an experimental focal cerebral ischemia, by middle cerebral artery occlusion for 90 minutes, followed by reperfusion for 48 hours, associated with an alcoholism model. Methods Fifty adult Wistar rats were used, subdivided into five experimental groups: control group (C): animals submitted to anesthesia only; sham group (S): animals submitted to complete simulation of the surgical procedure; ischemic group (I): animals submitted to focal cerebral ischemia for 90 minutes followed by reperfusion for 48 hours; alcoholic group (A): animals that received daily absolute ethanol diluted 20% in water for four weeks; and, ischemic and alcoholic group (I + A): animals receiving the same treatment as group A and, after four weeks, submitted to focal cerebral ischemia for 90 minutes, followed by reperfusion for 48 hours. The cerebellum samples were collected and immunohistochemical analysis of Caspase-9 protein and serum analysis by RT-PCR of microRNAs miR-21, miR-126 and miR155 were performed. Results The expression of Caspase-9 was higher in groups I, A and I + A. In the microRNAs analyses, miR-126 was higher in groups A and I + A, miR-155 was higher in groups I and I + A. Conclusions We conclude that apoptosis occurs in the cerebellar cortex, even if it is distant from the ischemic focus, and that microRNAs 126 and 155 show a correlation with cellular apoptosis in ischemic rats and those submitted to the chronic alcohol model.

https://doi.org/10.1590/0004-282x20190126 1287 downloads

Morphological and immunohistochemical analysis of proteins CASPASE 3 and XIAP in rats subjected to cerebral ischemia and chronic alcoholism
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Schiavoni, Vagner Sarraipo

; Silva, Jairo Pinheiro da

; Lizarte Neto, Fermino Sanches

; Assis, Múcio Luiz Cirino de

; Tazima, Maria de Fátima Galli Sorita

; Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Carlotti Jr, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luis Fernando

Acta Cirúrgica Brasileira

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Ago 2018, Volume 33 Nº 8 Páginas 652 - 663

Resumo: En | Texto: En | PDF: En

Abstract Purpose: To evaluate histopathological and ultrastructural changes and expression of proteins related to apoptosis CASPASE 3 and XIAP after experimental induction of temporary focal cerebral ischemia (90 minutes) due to obstruction of the middle cerebral artery in alcoholism model. Methods: Forty adult Wistar rats were used, subdivided into 5 experimental groups: control group (C); Sham group (S); Ischemic group (I); Alcoholic group (A); and Ischemic and Alcoholized group (I+A): animals submitted to the same treatment of group A and after four weeks were submitted to focal cerebral ischemia during 90 minutes, followed by reperfusion of 48 hours. Were processed for histopathological analysis and immunohistochemistry (for the protein expression of CASPASE -3 and XIAP). Results: Greater histopathological changes were observed in the animals of groups I and I+A in the three areas analyzed. The neuronal loss was higher in the medial striatum region of the animals of groups I and I + A. The protein expression of CASPASE -3 was higher than that of XIAP in the groups I and I + A for both proteins. Conclusion: The expression of XIAP was slightly higher where the histopathological changes and expression of CASPASE -3 was less evident.

https://doi.org/10.1590/s0102-865020180080000001 877 downloads

10.

High expression of XIAP and Bcl-2 may inhibit programmed cell death in glioblastomas
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Tirapelli, Daniela Pretti da Cunha

; Lustosa, Isis Lacrose

; Menezes, Sarah Bomfim

; Franco, Indira Maynart

; Rodrigues, Andressa Romualdo

; Peria, Fernanda Maris

; Marinho, Alexandre Magno da Nóbrega

; Serafini, Luciano Neder

; Carlotti Jr, Carlos Gilberto

; Tirapelli, Luís Fernando

Arquivos de Neuro-Psiquiatria

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Dez 2017, Volume 75 Nº 12 Páginas 875 - 880

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO O glioblastoma (GBM) é o glioma mais maligno e representa 29% de todos os tumores cerebrais. A tumorigênese está intimamente ligada à características adquiridas na via fisiológica de morte celular. Objetivo: Avaliar a expressão de genes anti-apoptóticos (XIAP e Bcl-2) e apoptóticos (citocromo C, a caspase 9, APAF-1), caspase 3 e SMAC/DIABLO, relacionados à apoptose, em 30 amostras de tecido de pacientes com glioblastoma. Métodos: A expressão gênica foi avaliada em trinta glioblastomas e comparada a dez amostras controles de substância branca por PCR em tempo real. Resultados e Conclusão: Houve maior nível de expressão de XIAP (p = 0,0032) e Bcl-2 (p = 0,0351) em comparação com as amostras controle, de cérebro normal. Estes resultados levantam a questão de que os genes Bcl-2 e XIAP podem ser responsáveis pela inibição da morte celular programada em glioblastomas, além disso, proporcionam informação adicional capaz de permitir o desenvolvimento de novas estratégias de terapia alvo.

ABSTRACT Glioblastoma (GBM) is the most malignant glioma and represents 29% of all brain tumors. Tumorigenesis is intimately connected with characteristics acquired in the physiologic pathway of cellular death. Objective: In the present study, the expression of anti-apoptotic (XIAP and Bcl-2) and apoptotic (cytochrome C, caspase 9, APAF-1), caspase 3 and the Smac/DIABLO genes related to the apoptosis pathway were evaluated in 30 samples of glioblastoma. Methods: The gene expression was evaluated in 30 glioblastomas (WHO grade IV) and compared to 10 white matter control samples with real-time PCR. Results and Conclusion: There were higher expressions of XIAP (p = 0.0032) and Bcl-2 (p = 0.0351) in the glioblastoma samples compared to the control samples of normal brain. These results raise the question of whether Bcl-2 and XIAP genes can be responsible for the inhibition of programmed cell death in glioblastomas. Moreover, they provide additional information capable of allowing the development of new target therapy strategies.

https://doi.org/10.1590/0004-282x20170156 1605 downloads

11.

High expression of anti-apoptotic genes in grade I and II meningiomas
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Tirapelli, Daniela Pretti da Cunha

; Menezes, Sarah Bomfim

; Franco, Indira Maynart

; Lustosa, Isis Lacrose

; Rodrigues, Andressa Romualdo

; Novais, Paulo Cézar

; Santiago, Antônio César Mendes

; Peria, Fernanda Maris

; Serafini, Luciano Neder

; Marinho, Alexandre Magno da Nóbrega

; Carlotti Jr, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

Arquivos de Neuro-Psiquiatria

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Abr 2017, Volume 75 Nº 4 Páginas 209 - 215

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO Um dos diferentes mecanismos genéticos envolvidos na carcinogênese de meningiomas é influenciado por interações entre proteínas que induzem e inibem a apoptose. Objetivos Avaliar a expressão de c-FLIP, XIAP, Bcl-2, caspase 3, 8 e 9, citocromo C, APAF 1 e Smac/DIABLO, genes relacionados com as vias da apoptose. Métodos A expressão gênica foi avaliada em trinta amostras de meningiomas (OMS grau I e II) e em dez amostras normais (de aracnóide) por PCR em tempo real. Resultados Os resultados mostraram maior expressão de genes antiapoptóticos em meningiomas quando comparados com controle, em contraste com a menor expressão de genes próapoptóticos. Conclusão Há um possível bloqueio na ativação de caspases através da via intrínseca da apoptose em meningiomas. O c-FLIP modula a caspase 8 e, desse modo, inibindo a sua ativação pela ausência de ligação com o receptor, há um bloqueio na ativação de FAS pela via extrínseca da apoptose.

ABSTRACT One of the different genetic mechanisms involved in the carcinogenesis of meningiomas is influenced by interactions between proteins that induce and inhibit apoptosis. Objective To evaluate the expression of c-FLIP, XIAP, Bcl-2, caspase 3, 8 and 9, cytochrome c, APAF 1 and Smac/DIABLO genes related to apoptosis pathways. Methods The gene expression was evaluated in 30 meningiomas (WHO grades I and II) and in 10 normal samples (from arachnoid tissue) through PCR-RT. Results The results showed higher expression of anti-apoptotic genes in meningiomas when compared to the control group, which had a low expression of pro-apoptotic genes. Conclusion There is a possible block in the activation of caspases through the intrinsic apoptosis pathway in meningiomas. c-FLIP modulates caspase 8 and, by inhibiting its activation due to the lack of connection with the receiver, there is a block to the FAS activation of apoptosis by its extrinsic pathway.

https://doi.org/10.1590/0004-282x20170027 1533 downloads

12.

Apoptosis in Bone Defect of Diabetic Rats Treated with Low Intensity Laser: Radiological and Immunohistochemical Approach
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da Silva, Jairo Pinheiro

; de Carvalho, Camila Albuquerque Mello

; Romualdo, Andressa Rodrigues

; Schiavoni, Vagner Sarraipo

; Barbosa, Marcello Henrique Nogueira

; de Moraes, Marcella Suélen Torrecilla

; Tirapelli, Daniela Petri Cunha

; Fazan, Valéria de Paula Sassoli

; Thomazini, José Antônio

; Tirapelli, Luis Fernando

International Journal of Morphology

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Mar 2017, Volume 35 Nº 1 Páginas 178 - 183

Resumo: En Es | Texto: En Es | PDF: En | PDF: Es

The aim of this study is evaluate the efficacy of 904 nm laser diode in bone regeneration in the bone defect in diabetic rats. Six groups of 10 male Wistar rats and 2 mm bone defects drilled on the left and right tibia were used. The diabetic animals were treated with streptozotocin (40 mg/kg, i.v.). We compared the diode laser doses of treatment of bone defects 50 w 4 J/cm and 100 w 4J/. The right tibia was used for immunohistochemical analysis with the apoptosis markers XIAP and Caspase-3 and the left tibia was submitted to computer tomography (CT). Caspase-3 marker showed greater amount of apoptosis in all the untreated groups compared to both laser treatments. There was no statistical significance for XIAP marker. CT scan showed improvement of bone defect area and volume in both laser treated groups, control and diabetic. Therefore the low intensity laser therapy was effective in accelerating bone repair in both, control and diabetic groups. It was evidenced in our study that diabetes influences bone repair negatively.

Los objetivos de este estudio fueron evaluar la eficacia del láser diodo de 904 nm en la regeneración ósea del defecto óseo en ratas diabéticas. Se utilizaron seis grupos de 10 ratas Wistar macho y se generó un defecto óseo de 2 mm en las tibias izquierda y derecha de los animales. El animal diabético fue generado con estreptozotocina (40 mg / kg, i.v.). Se compararon las dosis de tratamiento de los defectos óseos con láser de diodo de 50 w - 4 J / cm y 100w - 4 J /. La tibia derecha fue utilizada para el análisis inmunohistoquímico con los marcadores de apoptosis XIAP y Caspasa-3 y la tibia izquierda fue sometida a tomografía computarizada. El marcador caspasa-3 mostró mayor cantidad de apoptosis en todos los grupos no tratados en comparación con ambos tratamientos con láser. No hubo significación estadística para el marcador XIAP. La tomografía computarizada mostró una mejoría del área y el volumen de los defectos óseos en ambos grupos tratados con láser, control y diabéticos. Por lo tanto, la terapia con láser de baja intensidad fue eficaz en la aceleración de la reparación ósea tanto en los grupos control como en los diabéticos. Se evidenció en nuestro estudio que la diabetes afecta negativamente la reparación ósea.

1255 downloads

13.

Expression of NMDA receptor and microRNA-219 in rats submitted to cerebral ischemia associated with alcoholism
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Silva, Cristiane Iozzi

; Novais, Paulo Cézar

; Rodrigues, Andressa Romualdo

; Carvalho, Camila A.M.

; Colli, Benedicto Oscar

; Carlotti Jr., Carlos Gilberto

; Tirapelli, Luís Fernando

; Tirapelli, Daniela P.C.

Arquivos de Neuro-Psiquiatria

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Jan 2017, Volume 75 Nº 1 Páginas 30 - 35

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

RESUMO Algumas condições podem agravar os danos causados pelo processo isquêmico, tais como o consumo de álcool, e diversos mecanismos moleculares que estão envolvidos na fisiopatologia da isquemia cerebral, incluindo a expressão de neurotransmissores, e estes podem estar regulados por microRNAs. Objetivo: Avaliar a expressão de NMDA e do microRNA-219 no tecido cerebral e no sangue de animais submetidos à isquemia cerebral associada ao alcoolismo. Métodos: 50 ratos Wistar foram divididos em: controle, sham, isquêmico, alcoólico e isquêmico mais alcoólico. A expressão de microRNA-219 e de NMDA foram analisadas por PCR em tempo real. Resultados: Quando comparado com o grupo controle, o microRNA-219 no tecido cerebral foi menos expresso nos grupos isquêmico, alcoólico e associado. No sangue, este microRNA teve menor expressão no grupo alcoólico e no associado. Em relação à expressão do gene do NMDA, em tecido cerebral foi maior nos grupos isquêmico, alcoólico e no associado. Conclusão: Uma possível modulação de NMDA pelo microRNA-219 foi observada, com uma correlação inversa entre eles.

ABSTRACT Alcohol consumption aggravates injuries caused by ischemia. Many molecular mechanisms are involved in the pathophysiology of cerebral ischemia, including neurotransmitter expression, which is regulated by microRNAs. Objective: To evaluate the microRNA-219 and NMDA expression in brain tissue and blood of animals subjected to cerebral ischemia associated with alcoholism. Methods: Fifty Wistar rats were divided into groups: control, sham, ischemic, alcoholic, and ischemic plus alcoholic. The expression of microRNA-219 and NMDA were analyzed by real-time PCR. Results: When compared to the control group, the microRNA-219 in brain tissue was less expressed in the ischemic, alcoholic, and ischemic plus alcoholic groups. In the blood, this microRNA had lower expression in alcoholic and ischemic plus alcoholic groups. In the brain tissue the NMDA gene expression was greater in the ischemic, alcoholic, and ischemic plus alcoholic groups. Conclusion: A possible modulation of NMDA by microRNA-219 was observed with an inverse correlation between them.

https://doi.org/10.1590/0004-282x20160188 1543 downloads

14.

Expression profiles of eNOS, iNOS and microRNA-27b in the corpus cavernosum of rats submitted to chronic alcoholism and Diabetes mellitus
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Cunha, Joao Paulo da

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cezar

; Gattas, Daniela

; Carvalho, Camila Albuquerque Mello de

; Tirapelli, Daniela Pretti da Cunha

; Molina, Carlos Augusto Fernandes

; Tirapelli, Luis Fernando

; Tucci Junior, Silvio

Acta Cirúrgica Brasileira

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Jan 2017, Volume 32 Nº 1 Páginas 38 - 45

Resumo: En | Texto: En | PDF: En

Abstract Purpose: To evaluate the expression of endothelial and inducible NOS in addition to the miRNA-27b in the corpus cavernosum and peripheral blood of healthy rats, diabetic rats, alcoholic rats and rats with both pathologies. Methods: Forty eight Wistar rats were divided into four groups: control (C), alcoholic (A), diabetic (D) and alcoholic-diabetic (AD). Samples of the corpus cavernosum were prepared to study protein expressions of eNOS and iNOS by immunohistochemistry and expression of miRNA-27b in the corpus cavernosum and peripheral blood. Results: Immunohistochemistry for eNOS and iNOS showed an increase in cavernosal smooth muscle cells in the alcoholic, diabetic and alcoholic-diabetic groups when compared with the control group. Similarly, the mRNA levels for eNOS were increased in cavernosal smooth muscle (CSM) in the alcoholic, diabetic and alcoholic-diabetic groups and miRNA-27b were decreased in CSM in the alcoholic, diabetic and alcoholic-diabetic groups. Conclusion: The major new finding of our study was an impairment of relaxation of cavernosal smooth muscle in alcoholic, diabetic, and alcoholic-diabetic rats that involved a decrease in the nitric oxide pathway by endothelium-dependent mechanisms accompanied by a change in the corpus cavernosum contractile sensitivity.

https://doi.org/10.1590/s0102-865020170105 1446 downloads

15.

Morphological and immunohistochemical analysis of apoptosis in the cerebellum of rats subjected to focal cerebral ischemia with or without alcoholism model
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Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Rodrigues, Andressa Romualdo

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cézar

; Silva, Jairo Pinheiro

; Carlotti Júnior, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

Acta Cirúrgica Brasileira

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Set 2016, Volume 31 Nº 9 Páginas 629 - 637

Resumo: En | Texto: En | PDF: En

ABSTRACT PURPOSE: To evaluated histopathological changes, morphometric and expression of proteins CASPASE-3, BCL-2 and XIAP related to apoptosis in the cerebellum after induction of temporary focal cerebral ischemia followed by reperfusion, with or without a model of chronic alcoholism. METHODS: Fifty Wistar rats were used and divided into: control group (C), sham group (S), ischemic group (I), alcoholic group (A), and ischemic and alcoholic group (IA). The cerebellum samples collected were stained for histopathological and morphometric analysis and immunohistochemistry study. RESULTS: Histopathological changes were observed a greater degree in animals in groups A and IA. The morphometric study showed no difference in the amount of cells in the granular layer of the cerebellum between the groups. The expression of CASPASE-3 was higher than BCL-2 and XIAP in the groups A and IA. CONCLUSION: We observed correlation between histopathological changes and the occurrence of apoptosis in cerebellar cortex.

https://doi.org/10.1590/S0102-865020160090000009 2227 downloads

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