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1.
Int. j. morphol ; 38(6): 1639-1644, Dec. 2020. graf
Article in English | LILACS | ID: biblio-1134491

ABSTRACT

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.


Subject(s)
Animals , Rats , Apoptosis , Alcoholism/metabolism , Erectile Dysfunction/metabolism , Penis/physiopathology , Penis/chemistry , Immunohistochemistry , Rats, Wistar , MicroRNAs/analysis , MicroRNAs/genetics , MicroRNAs/metabolism , Disease Models, Animal , Alcoholism/physiopathology , Apoptosis Inducing Factor/analysis , Apoptosis Inducing Factor/genetics , Apoptosis Inducing Factor/metabolism , Real-Time Polymerase Chain Reaction , Erectile Dysfunction/physiopathology
2.
Acta cir. bras ; 35(3): e202000305, 2020. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1130627

ABSTRACT

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.

3.
Acta cir. bras ; 35(8): e202000805, 2020. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1130667

ABSTRACT

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.

4.
Acta cir. bras ; 32(1): 38-45, Jan. 2017. graf
Article in English | LILACS | ID: biblio-837673

ABSTRACT

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.


Subject(s)
Animals , Male , Rats , Penis/chemistry , MicroRNAs/analysis , Diabetes Mellitus, Experimental/metabolism , Alcoholism/metabolism , Nitric Oxide Synthase Type II/analysis , Nitric Oxide Synthase Type III/analysis , Penis/physiopathology , Immunohistochemistry , Rats, Wistar , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/physiopathology , Alcoholism/complications , Alcoholism/physiopathology
5.
Acta cir. bras ; 31(supl.1): 8-12, 2016. tab
Article in English | LILACS | ID: lil-779767

ABSTRACT

PURPOSE : Bladder augmentation is an effective surgical procedure for increasing bladder capacity and reducing pressure on the urinary system. It is indicated for patients with anomalies such as spina bifida, myelomeningocele, urethral valve and bladder exstrophy, who progress with low tolerance of medication. CASES : This was a retrospective study conducted on pediatric patients submitted to bladder augmentation from 2000 to 2011. RESULTS : 34 patients aged 4 to 17 years were submitted to bladder augmentation, 30 of them with an ileal loop and 4 with a ureter.A continent urinary shunt was performed in 16 patients, the Mitrofanoff conduit was associated in 15, and the Macedo technique was used in one. Mean follow-up was 34.35 months (1 to 122 months). Mean creatinine was 1.5 ng/ml (0.4 to 7.5 ng/ml) preoperatively and 1.78 ng/ml postoperatively. Three patients required a renal transplant during follow-up. There was improvement or resolution of vesicoureteral reflux in 83.5% of the kidneys on the right and in 75% on the left. Bladder capacity increased, on average, from 152.5 ml to 410 ml. The main complications were vesical lithiasis in 3 patients and conduit perforation in one. CONCLUSION : Bladder augmentation showed good results in this series, preserving renal function in most of the patients.


Subject(s)
Humans , Male , Female , Child, Preschool , Child , Adolescent , Postoperative Complications/etiology , Urologic Surgical Procedures/adverse effects , Urinary Bladder Diseases/surgery , Urologic Surgical Procedures/methods , Time Factors , Ureter/surgery , Urinary Bladder/surgery , Urinary Catheterization/adverse effects , Reproducibility of Results , Retrospective Studies , Risk Factors , Follow-Up Studies , Treatment Outcome
6.
Acta cir. bras ; 28(supl.1): 33-36, 2013. ilus
Article in English | LILACS | ID: lil-663889

ABSTRACT

PURPOSE: Fetal hydronephrosis is a frequent finding due to advances in prenatal ultrasonography. The definition of fetal and neonatal urinary tract obstruction is a very difficult task requiring confirmation of reduced renal function and hydronephrosis. In this study we followed a series of consecutive patients with intrauterine hydronephrosis that persisted during post-natal life. METHODS: 116 newborns with antenatal hydronephrosis diagnosed by ultrasound and submitted to a specific post-natal evaluative protocol with a follow-up period of 6 years. RESULTS: In 45 (38.8%) of 116 patients, ureteropelvic junction (UPJ) obstruction was confirmed and surgical correction of the UPJ obstruction was done in 19 patients. From 26 children who were initially submitted to non-surgical treatment, only 6 (23%) needed a surgical approach during follow up. Overall analysis showed that surgery was performed in 25 patients with UPJ obstruction, and the others 20 patients were kept under clinical observation, since normal renal function was confirmed by scintigraphy scans. CONCLUSION: Fetal hydronephrosis due to UPJ obstruction deserves careful postnatal evaluation. UPJ obstruction is the most frequent anomaly and its surgical treatment has very precise indications. The evaluative protocol was useful in identify patients that could be followed-up with a non-surgical approach.


OBJETIVO: Com a ampla utilização dos exames ultrassonográficos na avaliação pré-natal, é frequente o diagnóstico da hidronefrose fetal. A definição de obstrução do trato urinário no periodo pós-natal necessita da confirmação de redução da função renal além da hidronefrose. Neste estudo, acompanhamos uma série de pacientes consecutivos com hidronefrose intra-útero que persistiu no periodo pós-natal. MÉTODOS: 116 recém-nascidos com hidronefrose pré-natal diagnosticada pela ultrassonografia foram submetidos a protocolo específico de avaliação e companhados pelo periodo de 06 anos. RESULTADOS: Em 46 (38,8%) dos 116 pacientes foi confirmado o diagnóstico de estenose da junção ureteropiélica (JUP). Conforme os resultados do protocolo aplicado a correção cirúrgica da estenose da JUP foi realizada em 19 pacientes. Das 26 crianças encaminhadas inicialmente para observação clínica, apenas 6 (23%) necessitaram cirurgia durante o seguimento ambulatorial. Na análise geral, o procedimento cirúrgico para correção da estenose da JUP foi indicado em 25 pacientes. Nas outras 20 crianças não houve necessidade da realização da cirurgia. CONCLUSÃO: a hidronefrose fetal requer cuidadosa avaliação pós-natal. A estenose da junção pielo-ureteral é a anomalia mais frequente como causa da hidronefrose, e sua correção cirúrgica tem indicações precisas. O protocolo aplicado foi útil em diferenciar pacientes que não necessitaram cirurgia para tratamento da estenose da JUP.


Subject(s)
Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Pregnancy , Hydronephrosis/etiology , Ureteral Obstruction/complications , Hydronephrosis , Hydronephrosis/therapy , Hydronephrosis , Prospective Studies , Ultrasonography, Prenatal , Ureteral Obstruction , Ureteral Obstruction/surgery , Ureteral Obstruction , Urinary Tract/abnormalities , Urinary Tract , Urinary Tract
7.
Int. braz. j. urol ; 38(4): 536-543, July-Aug. 2012. ilus, tab
Article in English | LILACS | ID: lil-649448

ABSTRACT

OBJECTIVE: To investigate the VEGF expression and collagen deposition using a latex biomembrane as bladder replacement in rabbits. MATERIALS AND METHODS: After partial cystectomy, a patch of a non-vulcanized latex biomembrane (2 x 2 cm) was sewn to the bladder of rabbits with 5/0 monofilament polydioxanone sulfate sutures in a watertight manner. Groups of 5 animals were killed at 15, 45 and 90 days after surgery and the bladder was removed. Sections of 5µm were cut and stained with picrosirius-red in order to estimate the amount of extracellular matrix in the graft. To confirm the presence of VEGF in tissues, protein expression was determined by immunohistochemistry. RESULTS: No death, urinary leakage or graft extrusion occurred in any group. All bladders showed a spherical shape. A progressive reduction in the amount of collagen occurred in the graft area and was negatively and linearly correlated with time (p < 0.001). VEGF expression was higher in grafted areas when compared to controls at 15 and 45 days after surgery and decreased with time (p < 0.001). CONCLUSION: The latex biomembrane as a matrix for partial bladder replacement in rabbits promotes temporary collagen deposition and stimulates the angiogenic process.


Subject(s)
Animals , Male , Rabbits , Biocompatible Materials/therapeutic use , Collagen/analysis , Latex/therapeutic use , Urinary Bladder/surgery , Vascular Endothelial Growth Factor A/analysis , Collagen/metabolism , Disease Models, Animal , Immunohistochemistry , Membranes, Artificial , Regeneration , Time Factors , Vascular Endothelial Growth Factor A/metabolism
8.
Acta cir. bras ; 27(7): 477-481, jul. 2012. tab
Article in English | LILACS | ID: lil-640096

ABSTRACT

PURPOSE: To investigate the effect of lovastatin on renal ischemia followed by reperfusion. METHODS: Thirty one Wistar rats submitted to left renal ischemia for 60 minutes followed by contralateral nephrectomy were divided into two groups: A (n =17, control, no treatment), and B (n=14, lovastatin 15 mg/kg/day p.o. ten days before ischemia). The animals were sacrificed at the end of ischemia, after 24 hours and at seven days after reperfusion. Survival, serum urea and creatinine levels and renal mitochondrial function were evaluated. RESULTS: Mortality was 29.4% in group A and 0.7% in group B. Urea and creatinine levels were increased in both groups, but the values were significantly lower in group B. Mitochondrial function showed decoupling in 83.4% of group A, as opposed to 38.4/% of group B. CONCLUSIONS: The result shows a protective action of renal function by lovastatin administered before ischemia/reperfusion. Since most of the mitochondrial fraction presented membranes with the ability to maintain ATP production in group B, stabilization of the mitochondrial membrane should be considered as part of the protective action of lovastatin on renal function in ischemia/reperfusion.


OBJETIVO: Investigar a ação da lovastatina na isquemia renal seguida de reperfusão. MÉTODOS: Trinta e um ratos Wistar submetidos à isquemia renal esquerda durante 60 minutos, seguida da nefrectomia contralateral, foram distribuídos em dois grupos: A (n=17, controle, sem tratamento) e B (n=14, recebendo 15 mg/Kg/dia de lovastatina via oral), durante os dez dias que antecederam a isquemia. Os animais foram mortos ao final da isquemia, e com 24 horas e sete dias após a reperfusão. Foram avaliadas a sobrevida, os valores séricos de uréia e creatinina e a função mitocondrial renal. RESULTADOS: A mortalidade foi 29,4% no grupo A e 0,7% no grupo B. Os níveis de uréia e creatinina elevaram-se nos dois grupos, mas foram significativamente menores no grupo B. No grupo A a função mitocondrial renal ficou desacoplada em 83,4% dos ensaios, enquanto que no grupo B isto ocorreu em apenas 38,4% dos ensaios. CONCLUSÕES: Os resultados mostram que a administração de lovastatina antes do episódio de isquemia protege a função renal. No grupo B, como a maior parte da fração mitocondrial isolada apresentou função acoplada à produção de ATP, deve-se também considerar a estabilização da membrana mitocondrial como parte da ação protetora da lovastatina na função renal durante isquemia e reperfusão.


Subject(s)
Animals , Male , Rats , Hypolipidemic Agents/pharmacology , Kidney/drug effects , Lovastatin/pharmacology , Mitochondria, Liver/drug effects , Reperfusion Injury/drug therapy , Creatinine/blood , Kidney/blood supply , Kidney/physiopathology , Mitochondria, Liver/physiology , Nephrectomy , Rats, Wistar , Renal Circulation/drug effects , Renal Circulation/physiology , Reperfusion Injury/blood , Reperfusion Injury/physiopathology , Time Factors , Urea/blood
9.
Acta cir. bras ; 23(supl.1): 31-35, 2008. graf, tab
Article in English | LILACS | ID: lil-483120

ABSTRACT

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.


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.


Subject(s)
Animals , Male , Rats , Kidney/growth & development , Mitochondria/physiology , Reperfusion Injury/pathology , Warm Ischemia , Adaptation, Physiological/physiology , Disease Models, Animal , Kidney/physiopathology , Kidney/surgery , Nephrectomy , Organ Size , Rats, Wistar , Reperfusion Injury/physiopathology , Time Factors
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