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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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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

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

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.

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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

10.

Inibição da guanilato ciclase pelo azul de metileno no choque circulatório causado por pancreatite aguda necrosante: uma palavra de cuidado embasada em modelo suíno
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Baldo, Caroline Floreoto

; Capellini, Verena Kise

; Celotto, Andrea Carla

; Sônego, Fabiane

; Tirapelli, Luis Fernando

; Batalhão, Marcelo

; Cárnio, Evelin Capellari

; Santos, José Sebastião dos

; Evora, Paulo Roberto Barbosa

Revista do Colégio Brasileiro de Cirurgiões

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Dez 2013, Volume 40 Nº 6 Páginas 480 - 489

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

OBJETIVO: estudar o uso terapêutico do bloqueio da guanilato ciclase pelo azul de metileno em um modelo experimental de pancreatite aguda grave em suínos. MÉTODOS: a pancreatite aguda necrotizante foi induzida em porcos anestesiados por infusão ductal pancreática retrógrada de 1ml/kg de taurocolato de sódio a 5% e 8U/kg de enteroquinase. Três grupos foram estudados (n=5): controle (C), pancreatite (PA), "bolus" de azul seguido por pancreatite (AM+PA). Os dados incluíram enzimas séricas e do líquido abdominal, variáveis hemodinâmicas, hemogasometria arterial, volume de líquido abdominal, marcadores inflamatórios plasmáticos, nitrito/nitrato e mieloperoxidase e malondialdeído plasmático. Aplicou-se a análise de variância seguida do pós-teste de Bonferroni (p<0,05). RESULTADOS: os valores de amilase e lipase foram três e dez vezes mais elevados no grupo PA. A atividade da mieloperoxidase foi 50% superior no grupo PA. Os dados hemodinâmicos indicaram choque hipovolêmico precoce seguido de choque cardiogênico. Observou-se grave translocação de líquidos para a cavidade peritoneal. A nitrito/nitrato plasmática permaneceu inalterada. O grupo AM+PA teve aumento de cinco vezes do mieloperoxidase em comparação com o grupo C. CONCLUSÕES: a utilização de azul de metileno em suínos com pancreatite não demonstrou efeitos significativos sobre variáveis hemodinâmicas e inflamatórias. Seu uso terapêutico na pancreatite necro-hemorrágica pode ser inadequado e extremo cuidado deve ser tomado dado o aumento da peroxidação lipídica evidenciado pelo aumento dos valores do malondialdeído.

OBJECTIVE: To study the therapeutic application of guanylate cyclase inhibition by methylene blue in an experimental model of acute pancreatitis in pigs. METHODS: acute necrotizing pancreatitis was induced in anesthetized pigs by the retrograde infusion of 1 ml/kg of 5% sodium taurocholate and 8 U/kg enterokinase in the pancreatic duct. Three groups were studied (n = 5): control (C), pancreatitis (AP), and MB bolus followed by pancreatitis (MB+P). The data included serum and abdominal fluid enzymes, hemodynamic variables, arterial hemogasometry, abdominal fluid volume, inflammatory markers, plasma nitrite/nitrate (NOx), plasma myeloperoxidase (MPO) and plasma malondialdehyde (MDA). One- and two-way analysis of variance (ANOVA) was performed, followed by the Bonferroni test (p < 0.05). RESULTS: amylase and lipase were three and 10-fold higher in the AP group. Myeloperoxidase activity was 50% higher in the AP group. The hemodynamic data indicated early hypovolemic shock followed by cardiogenic shock. Severe fluid translocation to the peritoneal cavity was observed. Plasma NOx remained unchanged. The MB+P group had a five-fold increase in MDA compared with the C group. CONCLUSION: preemptive application of MB in pigs with AP demonstrated no significant effects on hemodynamic and inflammatory variables. The use of MB is inadequate in cases of exponential NO release, and extreme caution must be exercised, given the increase in lipid peroxidation based on the malondialdehyde dosage.

3074 downloads

11.

Expression of HSP70 in cerebral ischemia and neuroprotetive action of hypothermia and ketoprofen
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Tirapelli, Daniela Pretti da Cunha

; Carlotti Junior, Carlos Gilberto

; Leite, João Pereira

; Tirapelli, Luis Fernando

; Colli, Benedicto Oscar

Arquivos de Neuro-Psiquiatria

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Ago 2010, Volume 68 Nº 4 Páginas 592 - 596

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

Proteínas de choque térmico (HSPs) são chaperones moleculares que se ligam a outras proteínas para atravessar as membranas e encaminhá-las para locais específicos dentro de uma célula. Vários estímulos nocivos podem induzir a expressão de Hsp70, incluindo isquemia. Este estudo teve como objetivo investigar o padrão de expressão protéica (imunohistoquímica) e gênica (PCR em tempo real) de Hsp70 na isquemia cerebral focal experimental em ratos pela oclusão da artéria cerebral média durante 1 hora e o papel da neuroproteção com hipotermia (H) e cetoprofeno (C). O volume de infarto foi calculado através da análise morfométrica definido por cloreto de trifenil tetrazólio. Foi observado aumento na expressão proteína (p=0,0001) e gênica (p=0,0001) de Hsp70 nas áreas isquêmicas que foram reduzidas pela H (proteína e gene: p<0,05), C (proteína: p<0,001) e H+K (proteína: p<0,01 e gene: p<0,05). O aumento de Hsp70 na área isquêmica sugere que a neuroexcitotoxicidade mediada pela Hsp70 desempenha um papel importante na morte celular e que o efeito neuroprotetor tanto da H quanto do C está diretamente envolvido com a Hsp70.

Heat shock proteins (HSPs) are molecular chaperones that bind to other proteins to shepherd them across membranes and direct them to specific locations within a cell. Several injurious stimuli can induce Hsp70 expression, including ischemia. This study aimed to investigate the pattern of expression of protein (immunohistochemistry) and gene (real-time PCR) Hsp70 in experimental focal cerebral ischemia in rats by occlusion of the middle cerebral artery for 1 hour and the role of neuroprotection with hypothermia (H) and ketoprofen (K). The infarct volume was measured using morphometric analysis defined by triphenyl tetrazolium chloride. It was observed increases in the protein (p=0.0001) and gene (p=0.0001) Hsp70 receptor in the ischemic areas that were reduced by H (protein and gene: p<0.05), K (protein: p<0.001), and H+K (protein: p<0.01 and gene: p<0.05). The Hsp70 increases in the ischemic area suggests that the Hsp70-mediated neuroexcitotoxicity plays an important role in cell death and that the neuroprotective effect of both, H and K are directly involved with the Hsp70.

3551 downloads Citado 1 vez em SciELO

12.

Compensatory renal growth and mitochondrial function: the influence of warm ischemia and reperfusion
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Tucci Jr, Silvio

; Molina, Carlos Augusto Fernandes

; Cologna, Adauto José

; Suaid, Haylton Jorge

; Tirapelli, Luis Fernando

; Celini, Fábia Martins

; Martins, Antonio Carlos Pereira

Acta Cirúrgica Brasileira

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2008, Volume 23 Supl. 1 Páginas 31 - 35

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

OBJETIVO: Avaliar a influência da lesão de isquemia/reperfusão na hipertrofia renal compensatória (HRC) e na função mitocondrial. MÉTODOS: 45 ratos Wistar foram divididos em três grupos: Grupo Controle (GC) - 21 ratos submetidos apenas à laparotomia e sacrificados no 1º dia (6 ratos) e 7º dia pós-operatório (15 ratos) para avaliar o peso seco de ambos os rins e seu crescimento durante uma semana (6 ratos) e quantificar a função mitocondrial (9 ratos); Grupo 1 (G1) - 12 ratos submetidos à nefrectomia direita e sacrificados após 7 dias para análise da função mitocondrial renal (6 ratos) e peso renal seco (6 ratos); Grupo 2 (G2) - isquemia renal quente durante 60 minutos no rim esquerdo seguida da nefrectomia direita foi realizada em 12 ratos, que foram sacrificados após 7 dias para avaliação da função mitocondrial (6 ratos) e peso seco (6 ratos). RESULTADOS: peso seco (mg) do rim esquerdo no 7º dia: GC= 219±18; G1=281±23 e G2=338±39 (GCxG1 p<0,01; GCxG2 p<0,001; G1xG2 p<0,01). O estado 4 da função mitocondrial e a Razão de Controle Respiratório (RCR) foram semelhantes em todos os grupos (p>0,05). O estado 3 da respiração mitocondrial (mMO2/min/mg) no GC, G1 e G2 foi, respectivamente: 99±23, 132±22 e 88±44 (p<0,02; a única diferença estatística foi observada entre os grupos G1xG2 - p<0,05). CONCLUSÕES: após nefrectomia unilateral a HRC está associada ao aumento do estado 3 da respiração mitocondrial. A lesão de isquemia/reperfusão renal aumenta a HRC estimulada pela nefrectomia unilateral, mas este aumento parece independer da respiração mitocondrial.

PURPOSE: To evaluate the influence of ischemia/reperfusion injury on renal compensatory growth (CGR) and mitochondrial function. METHODS: Forty five Wistar rats were divided in 3 groups: Control Group (GC) - 21 rats were submitted to a sham laparotomy and sacrificed at 1st (6 rats) and 7th (15 rats) postoperative days to evaluate the dry weight of both kidneys and their growth during 1 week (6 rats) and to quantify mitochondrial respiration (9 rats); Group 1 (G1) - 12 rats underwent right nephrectomy and were sacrificed 7 days later for analysis of renal mitochondrial function (6 rats) and dry weight (6 rats). Group 2 (G2) - renal warm ischemia for 60 minutes followed by right nephrectomy was performed in 12 rats; they were sacrificed 7 days later to evaluate renal mitochondrial function (6 rats) and dry weight (6 rats). RESULTS: Dry weight (mg) of left kidneys at 7th day: GC - 219±18, G1 - 281±23 and G2 - 338±39 (GCxG1 p<0.01; GCxG2 p<0.001; G1xG2 p<0.01). State 4 mitochondrial respiration rate and respiratory control ratio (RCR) were similar in all groups (p>0.05). State 3 respirations (mM/min/mg) in GC, G1 and G2 was respectively: 99±23, 132±22 and 82±44 (p<0.02; the only statistical difference noted was between groups G1xG2 - p<0.05). CONCLUSIONS: Following unilateral nephrectomy CRG is associated with an increase in state 3 of mitochondrial respiration. Renal ischemia/reperfusion injury enhances the CRG provoked by unilateral nephrectomy but such enhancement seems independent on mitochondrial respiration.

2656 downloads Citado 1 vez em SciELO

13.

Renal ischemia and reperfusion injury: influence of chorpromazine on renal function and lipid peroxidation
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Tucci Junior, Silvio

; Carvalho, Roberto Marins de

; Celini, Fábia Martins

; Cologna, Adauto José

; Suaid, Haylton Jorge

; Tirapelli, Luis Fernando

; Martins, Antonio Carlos Pereira

Acta Cirúrgica Brasileira

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2008, Volume 23 Supl. 1 Páginas 42 - 46

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

OBJETIVO: avaliar a influência da clorpromazina (CPZ) na função renal e na peroxidação lipídica num modelo de lesão de isquemia/reperfusão renal em ratos. MÉTODOS: 48 ratos Wistar foram submetidos à laparotomia para clampamento da artéria renal esquerda durante 60 minutos, seguido da reperfusão e nefrectomia contralateral. Destes animais, 26 receberam 3 mg/kg de CPZ intravenosa 15 minutos antes da isquemia renal (G-E), sendo os 22 animais restantes utilizados como grupo controle isquêmico (G-C). Em 11 ratos do G-E e 8 do G-C foi feita a dosagem de uréia e creatinina sérica antes da isquemia renal e no 1º, 4º e 7º dia pós-operatório. Amostras de tecido do rim esquerdo foram obtidas aos 5 minutos (5 ratos de cada grupo) e 24 horas após reperfusão (9 G-C e 10 G-E) para dosagem de malondialdeído (MDA). Valores controle para níveis de MDA foram obtidos em rins retirados de 6 ratos normais. RESULTADOS: insuficiência renal aguda ocorreu em todos os animais mas os níveis séricos de uréia e creatinina foram significativamente menores no G-E (p<0,001). Os níveis de MDA apresentaram elevação acentuada na avaliação aos 5 minutos de reperfusão em ambos os grupos (p<0,05), retornando a valores próximos aos normais na avaliação com 24 horas. CONCLUSÃO: a CPZ conferiu proteção parcial da função renal aos rins submetidos à lesão de isquemia e reperfusão, aparentemente independente da inibição da peroxidação lipídica.

PURPOSE: To evaluate the influence of chlorpromazine (CPZ) on renal function and lipid peroxidation in a rat model of kidney ischemia/reperfusion injury. METHODS: Forty eight Wistar rats underwent a laparotomy for hilar clamping of left kidney with a bulldog clamp for 60 minutes followed by organ reperfusion and contralateral nephrectomy. Of these, 26 received 3mg/kg of CPZ intravenously 15 minutes before renal ischemia (G-E) while the remaining 22 were used as ischemic control group (G-C). Eleven rats of G-E and 8 of G-C were followed for blood urea nitrogen and creatinine determinations before renal ischemia and at 1st, 4th and 7th postoperative days. Samplings of left renal tissue were obtained at 5 minutes (5 rats from each group) and 24 hours (9 G-C and 10 of G-E) of reperfusion for malondialdehy (MDA) content determination. Controls of renal MDA content were determined in kidneys harvested from 6 additional normal rats. RESULTS: Acute renal failure occurred in all animals but levels of BUN and creatinine were significantly lower in G-E (p<0.001). MDA content rose strikingly at 5 minutes of reperfusion in both groups (p>0.05) and returned near to normal levels 24 hours later. CONCLUSION: CPZ conferred partial protection of renal function to kidneys submitted to ischemia/reperfusion injury that seems to be not dependent on inhibition of lipid peroxidation.

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14.

Cyclosporine action on kidneys of rats submitted to normothermic ischaemia and reperfusion
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Cologna, Adauto José

; Lima, Lucy Vieira da Silva

; Tucci Jr., Silvio

; Suaid, Haylton Jorge

; Reis, Rodolfo Borges

; Tirapelli, Luis Fernando

; Rodrigues Jr., Antônio Antunes

; Martins, Antonio Carlos Pereira

Acta Cirúrgica Brasileira

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2008, Volume 23 Supl. 1 Páginas 36 - 41

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

OBJETIVO: Verificar se a ciclosporina A (CsA) afeta a lesão provocada pela isquemia seguida de reperfusão em rins de ratos. MÉTODOS: Ratos Wistar machos foram separados em grupos: 1 grupo controle (GS) e 2 grupos experimentais (G1 e G2). O G1 foi dividido em G1A - isquemia de 60 minutos; e G1C - isquemia de 60 minutos associada a 20 mg/kg/dia de CsA. O G2 foi dividido em G2C semelhante ao G1, exceto pelo tempo de isquemia de 40 minutos. A isquemia renal foi provocada pelo pinçamento da artéria renal esquerda, após o procedimento, foi realizada a nefrectomia direita. Dosagem de uréia e creatinina séricos foram feitos no dia da cirurgia (D0) e no dia seguinte (D1). Após 24 horas de reperfusão o rim esquerdo foi removido para análise histológica. RESULTADOS: No GS não foram observados alterações de uréia e creatinina em D0 e D1, tampouco alterações estruturais. A comparação entre GS e G2C não mostrou diferença na função renal (p>0,05); o grau de lesão tecidual foi menor em G2C do que nos demais grupos experimentais. CONCLUSÃO: A CsA protegeu a função renal e reduziu a lesão tecidual em rins de ratos submetidos a 40 minutos de isquemia.

PURPOSE: To verify if rat kidneys lesioned by ischaemia followed by reperfusion are affected by cyclosporine A (CsA). METHODS: Male Wistar rats were randomly divided into three groups, control (GS) and experimental (G1 and G2). G1 was subdivided in two: G1A composed of animals submitted to 60 minutes ischaemia and G1C with the same ischaemic procedure associated to 20 mg/kg/day CsA. Group G2 was subdivided and treated in the same way as G1 except that ischaemia was applied only for 40 minutes. Clamping the left renal artery followed by right side nephrectomy induced kidney ischaemia. Serum urea and creatinine were quantified on the day of surgery (D0) and in the following day (D1). Twenty four hours after reperfusion the left kidney was removed and histologically analyzed. RESULTS: Group GS had normal values for urea and creatinine both on D0 and D1 and did not show structural alterations. Renal function was not significantly different when G2C was compared to GS (p>0.05). Tissue lesions were smaller in G2C than in the other groups. CONCLUSIONS: Renal function was protected by CsA, which also reduced tissue lesions in the kidneys of rats submitted to 40 minutes ischaemia.

2078 downloads Citado 2 vezes em SciELO

15.

Biochemical evaluation of focal non-reperfusion cerebral ischemia by middle cerebral artery occlusion in rats
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Colli, Benedicto Oscar

; Tirapelli, Daniela Pretti da Cunha

; Carlotti Jr, Carlos Gilberto

; Lopes, Luiza da Silva

; Tirapelli, Luis Fernando

Arquivos de Neuro-Psiquiatria

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2008, Volume 66 Nº 3b Páginas 725 - 730

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

Isquemia cerebral é um acontecimento importante na prática neurológica clínica e cirúrgica, uma vez que é uma das doenças que mais comprometem a espécie humana. No presente estudo 40 ratos adultos foram submetidos a períodos de isquemia focal de 30, 60 e 90 min e como controle foram utilizados animais do grupo sham. Foram analisados os níveis de ATP, malondialdeído e caspase-3. Nenhuma diferença significativa nas dosagens bioquímicas foram observadas entre os hemisférios cerebrais direito e esquerdo do mesmo animal em cada grupo experimental. Foi observada redução nos níveis de ATP após os três períodos de isquemia, em comparação com o grupo sham. Nenhum aumento significativo dos níveis de malondialdeído ou caspase-3 foi observado. Apesar das alterações significativas nos níveis ATP, os resultados indicaram viabilidade celular na região isquêmica como demonstrado pela baixa taxa de peroxidação lipídica e apoptose, achados que provavelmente estão relacionados com a falta de reperfusão.

Cerebral ischemia is an important event in clinical and surgical neurological practice since it is one of the diseases that most compromise the human species. In the present study 40 adult rats were submitted to periods of focal ischemia of 30, 60 and 90 min without reperfusion and animals submitted to a sham procedure were used as controls. We analyzed the levels of ATP, malondialdehyde and caspase-3. No significant differences in the biochemical measurements were observed between the right and left brain hemispheres of the same animal in each experimental group. Reduced ATP levels were observed after the three periods of ischemia compared to the sham group. No significant increase in malondialdehyde or caspase-3 levels was observed. Despite significant changes in ATP levels, the results indicated cell viability in the ischemic region as shown by the low rates of lipid peroxidation and apoptosis, findings probably related to the lack of reperfusion.

1904 downloads Citado 1 vez em SciELO

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