Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model

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

doi:10.1590/s0102-865020200030000005

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ORIGINAL ARTICLE, Experimental Surgery • Acta Cir. Bras. 35 (3) • 2020 • https://doi.org/10.1590/s0102-865020200030000005 linkcopy

Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model ¹

Authorship SCIMAGO INSTITUTIONS RANKINGS

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.

Ethanol; Erectile Dysfunction; Apoptosis; MicroRNAs; Rats

Authorship

person Rogério José de Azevedo Meirelles

schoolPhD, Department of Surgery and Anatomy, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (FMRP-USP), Ribeirao Preto-SP, Brazil. Acquisition of data.Universidade de São PauloBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (FMRP-USP), Ribeirao Preto-SP, Brazil. Acquisition of data.

0000-0001-9208-7120

person Fermino Sanches Lizarte Neto

schoolPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Technical procedures.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Technical procedures.

0000-0002-0046-7300

person Mucio Luiz de Assis Cirino

schoolFellow PhD degree, Postgraduate Program in Clinical Surgical, Department of Surgery and Anatomy, Surgical Clinic Program, FMRP-USP, Ribeirao Preto-SP, Brazil. Acquisition of data.USPBrazilRibeirao Preto, SP, BrazilFellow PhD degree, Postgraduate Program in Clinical Surgical, Department of Surgery and Anatomy, Surgical Clinic Program, FMRP-USP, Ribeirao Preto-SP, Brazil. Acquisition of data.

0000-0001-7872-8331

person Paulo Cezar Novais

schoolPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Postgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília (UNIMAR), Marilia-SP, Brazil. Histopathological examinations.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Postgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília (UNIMAR), Marilia-SP, Brazil. Histopathological examinations.schoolPostgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília, Marilia, SP, BrazilUniversidade de MaríliaBrazilMarilia, SP, BrazilPostgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília, Marilia, SP, Brazil

0000-0001-8372-6293

person Isabella Stracieri Gula

schoolGraduate student, School of Medicine, Universidade de Ribeirão Preto (UNAERP), Brazil. Acquisition of data.Universidade de Ribeirão PretoBrazilBrazilGraduate student, School of Medicine, Universidade de Ribeirão Preto (UNAERP), Brazil. Acquisition of data.

0000-0003-0797-9525

person Jairo Pinheiro da Silva

schoolPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Professor, Faculdade de Taquaritinga (FTGA), Brazil. Manuscript preparation.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Professor, Faculdade de Taquaritinga (FTGA), Brazil. Manuscript preparation.schoolFaculdade de Taquaritinga, BrazilFaculdade de TaquaritingaBrazilBrazilFaculdade de Taquaritinga, Brazil

0000-0002-5213-8525

person Maria de Fátima Galli Sorita Tazzima

schoolPhD, Assistant Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilRibeirao Preto, SP, BrazilPhD, Assistant Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.

0000-0002-5844-7310

person Valéria Paula Sassoli Fazan

schoolPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilRibeirao Preto, SP, BrazilPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.

0000-0003-1293-5308

person Marina Toledo Durand

schoolPhD, UNAERP, Ribeirao Preto-SP, Brazil. Manuscript writing.UNAERPBrazilRibeirao Preto, SP, BrazilPhD, UNAERP, Ribeirao Preto-SP, Brazil. Manuscript writing.

0000-0002-5625-4662

person Daniela Pretti da Cunha Tirapelli

schoolPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Final approval.USPBrazilRibeirao Preto, SP, BrazilPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Final approval.

0000-0002-1886-4988

person Camila Albuquerque Melo de Carvalho

schoolPhD, Department of Surgery and Anatomy, FMRP-USP, and Professor, Centro Universitário Barão de Mauá, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilBrazilPhD, Department of Surgery and Anatomy, FMRP-USP, and Professor, Centro Universitário Barão de Mauá, Ribeirao Preto-SP, Brazil. Manuscript writing.schoolCentro Universitário Barão de Mauá, Ribeirao Preto, SP, BrazilCentro Universitário Barão de MauáBrazilRibeirao Preto, SP, BrazilCentro Universitário Barão de Mauá, Ribeirao Preto, SP, Brazil

0000-0003-2325-2844

person Bruno César Schimming

schoolPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Associate Professor, Department of Bioscience, Universidade Estadual Paulista (UNESP), Botucatu-SP, Brazil. Critical revision.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Associate Professor, Department of Bioscience, Universidade Estadual Paulista (UNESP), Botucatu-SP, Brazil. Critical revision.schoolDepartment of Bioscience, Universidade Estadual Paulista, Botucatu, SP, BrazilUniversidade Estadual PaulistaBrazilBotucatu, SP, BrazilDepartment of Bioscience, Universidade Estadual Paulista, Botucatu, SP, Brazil

0000-0002-0837-9612

person Carlos Augusto Fernandes Molina

0000-0002-5118-778X

person Silvio Tucci Junior

0000-0002-7656-0181

person Luis Fernando Tirapelli

0000-0002-2251-1106

Correspondence: Luis Fernando Tirapelli Avenida Bandeirantes, 3900 14049-900 Ribeirão Preto-SP Brasil Tel.: (55 16)3315-3305
emaillab.biomol.cirurgia@fmrp.usp.br

Conflict of interest: none

SCIMAGO INSTITUTIONS RANKINGS

PhD, Department of Surgery and Anatomy, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (FMRP-USP), Ribeirao Preto-SP, Brazil. Acquisition of data.Universidade de São PauloBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (FMRP-USP), Ribeirao Preto-SP, Brazil. Acquisition of data.

PhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Technical procedures.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Technical procedures.

Fellow PhD degree, Postgraduate Program in Clinical Surgical, Department of Surgery and Anatomy, Surgical Clinic Program, FMRP-USP, Ribeirao Preto-SP, Brazil. Acquisition of data.USPBrazilRibeirao Preto, SP, BrazilFellow PhD degree, Postgraduate Program in Clinical Surgical, Department of Surgery and Anatomy, Surgical Clinic Program, FMRP-USP, Ribeirao Preto-SP, Brazil. Acquisition of data.

PhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Postgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília (UNIMAR), Marilia-SP, Brazil. Histopathological examinations.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Postgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília (UNIMAR), Marilia-SP, Brazil. Histopathological examinations.

Postgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília, Marilia, SP, BrazilUniversidade de MaríliaBrazilMarilia, SP, BrazilPostgarduate Program in Structural and Functional Interactions in Rehabiliattion, Universidade de Marília, Marilia, SP, Brazil

Graduate student, School of Medicine, Universidade de Ribeirão Preto (UNAERP), Brazil. Acquisition of data.Universidade de Ribeirão PretoBrazilBrazilGraduate student, School of Medicine, Universidade de Ribeirão Preto (UNAERP), Brazil. Acquisition of data.

PhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Professor, Faculdade de Taquaritinga (FTGA), Brazil. Manuscript preparation.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Professor, Faculdade de Taquaritinga (FTGA), Brazil. Manuscript preparation.

Faculdade de Taquaritinga, BrazilFaculdade de TaquaritingaBrazilBrazilFaculdade de Taquaritinga, Brazil

PhD, Assistant Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilRibeirao Preto, SP, BrazilPhD, Assistant Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.

PhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilRibeirao Preto, SP, BrazilPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Manuscript writing.

PhD, UNAERP, Ribeirao Preto-SP, Brazil. Manuscript writing.UNAERPBrazilRibeirao Preto, SP, BrazilPhD, UNAERP, Ribeirao Preto-SP, Brazil. Manuscript writing.

PhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Final approval.USPBrazilRibeirao Preto, SP, BrazilPhD, Associate Professor, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, Brazil. Final approval.

PhD, Department of Surgery and Anatomy, FMRP-USP, and Professor, Centro Universitário Barão de Mauá, Ribeirao Preto-SP, Brazil. Manuscript writing.USPBrazilBrazilPhD, Department of Surgery and Anatomy, FMRP-USP, and Professor, Centro Universitário Barão de Mauá, Ribeirao Preto-SP, Brazil. Manuscript writing.

Centro Universitário Barão de Mauá, Ribeirao Preto, SP, BrazilCentro Universitário Barão de MauáBrazilRibeirao Preto, SP, BrazilCentro Universitário Barão de Mauá, Ribeirao Preto, SP, Brazil

PhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Associate Professor, Department of Bioscience, Universidade Estadual Paulista (UNESP), Botucatu-SP, Brazil. Critical revision.USPBrazilRibeirao Preto, SP, BrazilPhD, Department of Surgery and Anatomy, FMRP-USP, Ribeirao Preto-SP, and Associate Professor, Department of Bioscience, Universidade Estadual Paulista (UNESP), Botucatu-SP, Brazil. Critical revision.

Department of Bioscience, Universidade Estadual Paulista, Botucatu, SP, BrazilUniversidade Estadual PaulistaBrazilBotucatu, SP, BrazilDepartment of Bioscience, Universidade Estadual Paulista, Botucatu, SP, Brazil

Figures

Figures (8)

Thumbnail

Figure 1 Cross-sectional photomicrograph of the rat penis where the two corpus cavernosum (CC) covered by the tunica albuginea ( arrow ) can be identified and contain a large number of cavernous ( blank ) spaces within it; ventrally the urethra (U) inside the spongy body and the dorsal deep vein of the penis (VDP). Masson’s trichrome, x50.

Thumbnail

Figure 2 Photomicrography showing detail of some connective trabeculae composed mainly of collagen fibers (CF) in blue and by subendothelial smooth muscle ( thinner arrows ). In the center a large cavernous space (EC) containing red blood cells inside ( red ) and endothelial ( thick arrow ). Masson’s trichrome, x400.

Thumbnail

Figure 3 Average area occupied by the smooth muscle fibers of the corpus cavernous of each animal from A and C groups (Two- way ANOVA, p=0.0002).

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Figure 4 Average area occupied by collagen fibers corpus cavernosum of the C and A groups (Mann-Whitney test, p=0.1988).

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Figure 5 Average area occupied by gaps or cavernous spaces of the corpus cavernosum in groups C and A (Mann-Whitney test, p=0.1583).

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Figure 6 Average of the protein expression of caspase 3 in the corpus cavernosum of C and A groups (Two-way ANOVA, p<0.0001).

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Figure 7 Gene expression of miRNA -21 in the serum in groups C and A (Mann- Whitney test, p=0.2135).

Thumbnail

Figure 8 Gene expression of miRNA -21 in the corpus cavernosum tissue in groups C and A (Mann- Whitney test, p=0.864).

imageFigure 1 Cross-sectional photomicrograph of the rat penis where the two corpus cavernosum (CC) covered by the tunica albuginea ( arrow ) can be identified and contain a large number of cavernous ( blank ) spaces within it; ventrally the urethra (U) inside the spongy body and the dorsal deep vein of the penis (VDP). Masson’s trichrome, x50. open_in_new

imageFigure 2 Photomicrography showing detail of some connective trabeculae composed mainly of collagen fibers (CF) in blue and by subendothelial smooth muscle ( thinner arrows ). In the center a large cavernous space (EC) containing red blood cells inside ( red ) and endothelial ( thick arrow ). Masson’s trichrome, x400. open_in_new

imageFigure 3 Average area occupied by the smooth muscle fibers of the corpus cavernous of each animal from A and C groups (Two- way ANOVA, p=0.0002). open_in_new

imageFigure 4 Average area occupied by collagen fibers corpus cavernosum of the C and A groups (Mann-Whitney test, p=0.1988). open_in_new

imageFigure 5 Average area occupied by gaps or cavernous spaces of the corpus cavernosum in groups C and A (Mann-Whitney test, p=0.1583). open_in_new

imageFigure 6 Average of the protein expression of caspase 3 in the corpus cavernosum of C and A groups (Two-way ANOVA, p<0.0001). open_in_new

imageFigure 7 Gene expression of miRNA -21 in the serum in groups C and A (Mann- Whitney test, p=0.2135). open_in_new

imageFigure 8 Gene expression of miRNA -21 in the corpus cavernosum tissue in groups C and A (Mann- Whitney test, p=0.864). open_in_new

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Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model 1

Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model ¹