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Published in: Journal of Assisted Reproduction and Genetics 12/2016

01-12-2016 | Reproductive Physiology and Disease

Alterations in the proliferative/apoptotic equilibrium in semen of adolescents with varicocele

Authors: Larissa Berloffa Belardin, Paula Toni Del Giudice, Mariana Camargo, Paula Intasqui, Mariana Pereira Antoniassi, Ricardo Pimenta Bertolla, Agnaldo Pereira Cedenho

Published in: Journal of Assisted Reproduction and Genetics | Issue 12/2016

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Abstract

Purpose

To verify if the presence of varicocele (grades II and III) with and without seminal alterations, using the 5th centile cutoff values in table A1.1 of the World Health Organization (WHO, 2010) manual, alters the seminal plasma levels of proteins DNASE1 (deoxyribonuclease-1) and IGFBP7 (Insulin-like growth factor-binding protein 7), which are related to apoptosis regulation and cell proliferation, respectively, demonstrating that these proteins are important for correct spermatogenesis.

Methods

This cross sectional study was performed at the Sao Paulo Federal University Paulo between May 2014 and April 2016. A total of 61 male adolescents were included in this study, of which 20 controls without varicocele (C), 22 with varicocele and normal semen analysis (VNS) and 19 with varicocele and altered semen analysis (VAS). Seminal plasma from each patient was used for Western blotting analysis of individual protein levels. Values of each protein were normalized to a testicular housekeeping protein (PARK7—protein deglycase DJ-1).

Results

Levels of IGFBP7 protein are increased in varicocele. Levels of DNASE1 are progressively decreased in varicocele (lower in varicocele and normal semen analysis, lowest in varicocele and altered semen analysis) when compared to adolescents without varicocele. DNASE1 levels are positively correlated with sperm concentration and morphology (correlation values of 0.400 and 0.404, respectively; p values of 0.001 and 0.001, respectively).

Conclusion

In conclusion, in adolescents, seminal plasma levels of IGFBP7, responsible for proliferative activity, are increased in varicocele grades II and III, and DNASE1, responsible for apoptosis regulation, are lower in varicocele, lowest in varicocele and low semen quality. These proteins demonstrate molecular alterations brought upon by varicocele. Moreover, DNASE1 is capable of discriminating a varicocele that causes alterations to semen quality from one that does not. We propose that the initial response of varicocele is to increase proliferative activity which, if followed by regulation of apoptosis, may lead to the ejaculation of a population of sperm that are in accordance with WHO cutoff values but, in the presence of dysregulated apoptosis, leads to lower sperm concentration and morphology.
Literature
1.
go back to reference Gat Y, Bachar GN, Zukerman Z, Belenky A, Gornish M. Varicocele: a bilateral disease. Fertil Steril. 2004;81:424–9.CrossRefPubMed Gat Y, Bachar GN, Zukerman Z, Belenky A, Gornish M. Varicocele: a bilateral disease. Fertil Steril. 2004;81:424–9.CrossRefPubMed
2.
go back to reference Benoff S, Gilbert BR. Varicocele and male infertility: part I. Preface Hum Reprod Update. 2001;7:47–54.CrossRefPubMed Benoff S, Gilbert BR. Varicocele and male infertility: part I. Preface Hum Reprod Update. 2001;7:47–54.CrossRefPubMed
3.
go back to reference Hauser R, Paz G, Botchan A, Yogev L, Yavetz H. Varicocele: effect on sperm functions. Hum Reprod Update. 2001;7:482–5.CrossRefPubMed Hauser R, Paz G, Botchan A, Yogev L, Yavetz H. Varicocele: effect on sperm functions. Hum Reprod Update. 2001;7:482–5.CrossRefPubMed
4.
go back to reference Smith R, Kaune H, Parodi D, Madariaga M, Rios R, Morales I, et al. Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. Hum Reprod Oxf Engl. 2006;21:986–93.CrossRef Smith R, Kaune H, Parodi D, Madariaga M, Rios R, Morales I, et al. Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. Hum Reprod Oxf Engl. 2006;21:986–93.CrossRef
5.
6.
go back to reference Gorelick JI, Goldstein M. Loss of fertility in men with varicocele. Fertil Steril. 1993;59:613–6.CrossRefPubMed Gorelick JI, Goldstein M. Loss of fertility in men with varicocele. Fertil Steril. 1993;59:613–6.CrossRefPubMed
7.
go back to reference Saleh RA, Agarwal A, Sharma RK, Said TM, Sikka SC, Thomas AJ. Evaluation of nuclear DNA damage in spermatozoa from infertile men with varicocele. Fertil Steril. 2003;80:1431–6.CrossRefPubMed Saleh RA, Agarwal A, Sharma RK, Said TM, Sikka SC, Thomas AJ. Evaluation of nuclear DNA damage in spermatozoa from infertile men with varicocele. Fertil Steril. 2003;80:1431–6.CrossRefPubMed
8.
go back to reference Zini A, Blumenfeld A, Libman J, Willis J. Beneficial effect of microsurgical varicocelectomy on human sperm DNA integrity. Hum Reprod Oxf Engl. 2005;20:1018–21.CrossRef Zini A, Blumenfeld A, Libman J, Willis J. Beneficial effect of microsurgical varicocelectomy on human sperm DNA integrity. Hum Reprod Oxf Engl. 2005;20:1018–21.CrossRef
9.
go back to reference Cozzolino DJ, Lipshultz LI. Varicocele as a progressive lesion: positive effect of varicocele repair. Hum Reprod Update. 2001;7:55–8.CrossRefPubMed Cozzolino DJ, Lipshultz LI. Varicocele as a progressive lesion: positive effect of varicocele repair. Hum Reprod Update. 2001;7:55–8.CrossRefPubMed
10.
go back to reference The influence of varicocele on parameters of fertility in a large group of men presenting to infertility clinics. World Health Organization. Fertil Steril 1992;57:1289–93. The influence of varicocele on parameters of fertility in a large group of men presenting to infertility clinics. World Health Organization. Fertil Steril 1992;57:1289–93.
11.
go back to reference Diamond D. Adolescent versus adult varicoceles—how do evaluation and management differ? J Urol. 2009;181:2418–9.CrossRefPubMed Diamond D. Adolescent versus adult varicoceles—how do evaluation and management differ? J Urol. 2009;181:2418–9.CrossRefPubMed
12.
go back to reference Practice Committee of the American Society for Reproductive Medicine, Society for Male Reproduction and Urology. Report on varicocele and infertility: a committee opinion. Fertil Steril 2014;102:1556–60. Practice Committee of the American Society for Reproductive Medicine, Society for Male Reproduction and Urology. Report on varicocele and infertility: a committee opinion. Fertil Steril 2014;102:1556–60.
13.
go back to reference Bertolla RP, Cedenho AP, Hassun Filho PA, Lima SB, Ortiz V, Srougi M. Sperm nuclear DNA fragmentation in adolescents with varicocele. Fertil Steril. 2006;85:625–8.CrossRefPubMed Bertolla RP, Cedenho AP, Hassun Filho PA, Lima SB, Ortiz V, Srougi M. Sperm nuclear DNA fragmentation in adolescents with varicocele. Fertil Steril. 2006;85:625–8.CrossRefPubMed
14.
go back to reference Lacerda JI, Del Giudice PT, da Silva BF, Nichi M, Fariello RM, Fraietta R, et al. Adolescent varicocele: improved sperm function after varicocelectomy. Fertil Steril. 2011;95:994–9.CrossRefPubMed Lacerda JI, Del Giudice PT, da Silva BF, Nichi M, Fariello RM, Fraietta R, et al. Adolescent varicocele: improved sperm function after varicocelectomy. Fertil Steril. 2011;95:994–9.CrossRefPubMed
15.
go back to reference Agarwal A, Deepinder F, Cocuzza M, Agarwal R, Short RA, Sabanegh E, et al. Efficacy of varicocelectomy in improving semen parameters: new meta-analytical approach. Urology. 2007;70:532–8.CrossRefPubMed Agarwal A, Deepinder F, Cocuzza M, Agarwal R, Short RA, Sabanegh E, et al. Efficacy of varicocelectomy in improving semen parameters: new meta-analytical approach. Urology. 2007;70:532–8.CrossRefPubMed
16.
go back to reference Gentile DP, Cockett AT. The effect of varicocelectomy on testicular volume in 89 infertile adult males with varicoceles. Fertil Steril. 1992;58:209–11.CrossRefPubMed Gentile DP, Cockett AT. The effect of varicocelectomy on testicular volume in 89 infertile adult males with varicoceles. Fertil Steril. 1992;58:209–11.CrossRefPubMed
17.
go back to reference Yamamoto M, Hibi H, Katsuno S, Miyake K. Effects of varicocelectomy on testis volume and semen parameters in adolescents: a randomized prospective study. Nagoya J Med Sci. 1995;58:127–32.PubMed Yamamoto M, Hibi H, Katsuno S, Miyake K. Effects of varicocelectomy on testis volume and semen parameters in adolescents: a randomized prospective study. Nagoya J Med Sci. 1995;58:127–32.PubMed
18.
go back to reference Cocuzza M, Cocuzza MA, Bragais FMP, Agarwal A. The role of varicocele repair in the new era of assisted reproductive technology. Clin São Paulo Braz. 2008;63:395–404.CrossRef Cocuzza M, Cocuzza MA, Bragais FMP, Agarwal A. The role of varicocele repair in the new era of assisted reproductive technology. Clin São Paulo Braz. 2008;63:395–404.CrossRef
19.
go back to reference Del Giudice PT, da Silva BF, Lo Turco EG, Fraietta R, Spaine DM, Santos LFA, et al. Changes in the seminal plasma proteome of adolescents before and after varicocelectomy. Fertil Steril. 2013;100:667–72.CrossRefPubMed Del Giudice PT, da Silva BF, Lo Turco EG, Fraietta R, Spaine DM, Santos LFA, et al. Changes in the seminal plasma proteome of adolescents before and after varicocelectomy. Fertil Steril. 2013;100:667–72.CrossRefPubMed
20.
go back to reference Zylbersztejn DS, Andreoni C, Del Giudice PT, Spaine DM, Borsari L, Souza GHMF, et al. Proteomic analysis of seminal plasma in adolescents with and without varicocele. Fertil Steril. 2013;99:92–8.CrossRefPubMed Zylbersztejn DS, Andreoni C, Del Giudice PT, Spaine DM, Borsari L, Souza GHMF, et al. Proteomic analysis of seminal plasma in adolescents with and without varicocele. Fertil Steril. 2013;99:92–8.CrossRefPubMed
21.
go back to reference Del Giudice PT, Belardin LB, Camargo M, Zylbersztejn DS, Carvalho VM, Cardozo KHM, et al. Determination of testicular function in adolescents with varicocoele—a proteomics approach. Andrology. 2016;4:447–55.CrossRefPubMed Del Giudice PT, Belardin LB, Camargo M, Zylbersztejn DS, Carvalho VM, Cardozo KHM, et al. Determination of testicular function in adolescents with varicocoele—a proteomics approach. Andrology. 2016;4:447–55.CrossRefPubMed
22.
go back to reference Samejima K, Earnshaw WC. Trashing the genome: the role of nucleases during apoptosis. Nat Rev Mol Cell Biol. 2005;6:677–88.CrossRefPubMed Samejima K, Earnshaw WC. Trashing the genome: the role of nucleases during apoptosis. Nat Rev Mol Cell Biol. 2005;6:677–88.CrossRefPubMed
23.
go back to reference Pollak M. Insulin and insulin-like growth factor signalling in neoplasia. Nat Rev Cancer. 2008;8:915–28.CrossRefPubMed Pollak M. Insulin and insulin-like growth factor signalling in neoplasia. Nat Rev Cancer. 2008;8:915–28.CrossRefPubMed
25.
go back to reference Dubin L, Amelar RD. Varicocele size and results of varicocelectomy in selected subfertile men with varicocele. Fertil Steril. 1970;21:606–9.CrossRefPubMed Dubin L, Amelar RD. Varicocele size and results of varicocelectomy in selected subfertile men with varicocele. Fertil Steril. 1970;21:606–9.CrossRefPubMed
26.
go back to reference Mori MM, Bertolla RP, Fraietta R, Ortiz V, Cedenho AP. Does varicocele grade determine extent of alteration to spermatogenesis in adolescents? Fertil Steril. 2008;90:1769–73.CrossRefPubMed Mori MM, Bertolla RP, Fraietta R, Ortiz V, Cedenho AP. Does varicocele grade determine extent of alteration to spermatogenesis in adolescents? Fertil Steril. 2008;90:1769–73.CrossRefPubMed
27.
go back to reference World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. Cambrige University Press; 2010. World Health Organization. Laboratory manual for the examination of human semen and sperm-cervical mucus interaction. Cambrige University Press; 2010.
28.
go back to reference Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985;150:76–85.CrossRefPubMed Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985;150:76–85.CrossRefPubMed
29.
go back to reference Mori MM, Cedenho AP, Koifman S, Srougi M. Sperm characteristics in a sample of healthy adolescents in São Paulo. Brazil Cad Saúde Pública. 2002;18:525–30.CrossRefPubMed Mori MM, Cedenho AP, Koifman S, Srougi M. Sperm characteristics in a sample of healthy adolescents in São Paulo. Brazil Cad Saúde Pública. 2002;18:525–30.CrossRefPubMed
30.
go back to reference Milardi D, Grande G, Sacchini D, Astorri AL, Pompa G, Giampietro A, et al. Male fertility and reduction in semen parameters: a single tertiary-care center experience. Int J Endocrinol. 2012;2012:649149.CrossRefPubMedPubMedCentral Milardi D, Grande G, Sacchini D, Astorri AL, Pompa G, Giampietro A, et al. Male fertility and reduction in semen parameters: a single tertiary-care center experience. Int J Endocrinol. 2012;2012:649149.CrossRefPubMedPubMedCentral
31.
go back to reference Camargo M, Intasqui P, Bertolla RP. Proteomic profile of seminal plasma in adolescents and adults with treated and untreated varicocele. Asian J Androl. 2016;18:194–201.CrossRefPubMed Camargo M, Intasqui P, Bertolla RP. Proteomic profile of seminal plasma in adolescents and adults with treated and untreated varicocele. Asian J Androl. 2016;18:194–201.CrossRefPubMed
32.
go back to reference World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. 2010. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th ed. 2010.
33.
go back to reference Gerena RL, Irikura D, Urade Y, Eguchi N, Chapman DA, Killian GJ. Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase. Biol Reprod. 1998;58:826–33.CrossRefPubMed Gerena RL, Irikura D, Urade Y, Eguchi N, Chapman DA, Killian GJ. Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase. Biol Reprod. 1998;58:826–33.CrossRefPubMed
34.
go back to reference D’Amours O, Frenette G, Bordeleau L-J, Allard N, Leclerc P, Blondin P, et al. Epididymosomes transfer epididymal sperm binding protein 1 (ELSPBP1) to dead spermatozoa during epididymal transit in bovine. Biol Reprod. 2012;87:94.CrossRefPubMed D’Amours O, Frenette G, Bordeleau L-J, Allard N, Leclerc P, Blondin P, et al. Epididymosomes transfer epididymal sperm binding protein 1 (ELSPBP1) to dead spermatozoa during epididymal transit in bovine. Biol Reprod. 2012;87:94.CrossRefPubMed
36.
go back to reference Utleg AG, Yi EC, Xie T, Shannon P, White JT, Goodlett DR, et al. Proteomic analysis of human prostasomes. Prostate. 2003;56:150–61.CrossRefPubMed Utleg AG, Yi EC, Xie T, Shannon P, White JT, Goodlett DR, et al. Proteomic analysis of human prostasomes. Prostate. 2003;56:150–61.CrossRefPubMed
37.
go back to reference Akaogi K, Okabe Y, Funahashi K, Yoshitake Y, Nishikawa K, Yasumitsu H, et al. Cell adhesion activity of a 30-kDa major secreted protein from human bladder carcinoma cells. Biochem Biophys Res Commun. 1994;198:1046–53.CrossRefPubMed Akaogi K, Okabe Y, Funahashi K, Yoshitake Y, Nishikawa K, Yasumitsu H, et al. Cell adhesion activity of a 30-kDa major secreted protein from human bladder carcinoma cells. Biochem Biophys Res Commun. 1994;198:1046–53.CrossRefPubMed
38.
go back to reference Oh Y, Nagalla SR, Yamanaka Y, Kim HS, Wilson E, Rosenfeld RG. Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. Recombinant human mac25 protein specifically binds IGF-I and -II. J Biol Chem. 1996;271:30322–5.CrossRefPubMed Oh Y, Nagalla SR, Yamanaka Y, Kim HS, Wilson E, Rosenfeld RG. Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. Recombinant human mac25 protein specifically binds IGF-I and -II. J Biol Chem. 1996;271:30322–5.CrossRefPubMed
39.
go back to reference Yamauchi T, Umeda F, Masakado M, Isaji M, Mizushima S, Nawata H. Purification and molecular cloning of prostacyclin-stimulating factor from serum-free conditioned medium of human diploid fibroblast cells. Biochem J. 1994;303:591–8.CrossRefPubMedPubMedCentral Yamauchi T, Umeda F, Masakado M, Isaji M, Mizushima S, Nawata H. Purification and molecular cloning of prostacyclin-stimulating factor from serum-free conditioned medium of human diploid fibroblast cells. Biochem J. 1994;303:591–8.CrossRefPubMedPubMedCentral
40.
go back to reference Agarwal A, Hamada A, Esteves SC. Insight into oxidative stress in varicocele-associated male infertility: part 1. Nat Rev Urol. 2012;9:678–90.CrossRefPubMed Agarwal A, Hamada A, Esteves SC. Insight into oxidative stress in varicocele-associated male infertility: part 1. Nat Rev Urol. 2012;9:678–90.CrossRefPubMed
41.
go back to reference Hamada A, Esteves SC, Agarwal A. Insight into oxidative stress in varicocele-associated male infertility: part 2. Nat Rev Urol. 2013;10:26–37.CrossRefPubMed Hamada A, Esteves SC, Agarwal A. Insight into oxidative stress in varicocele-associated male infertility: part 2. Nat Rev Urol. 2013;10:26–37.CrossRefPubMed
42.
go back to reference Aitken RJ, Clarkson JS. Cellular basis of defective sperm function and its association with the genesis of reactive oxygen species by human spermatozoa. J Reprod Fertil. 1987;81:459–69.CrossRefPubMed Aitken RJ, Clarkson JS. Cellular basis of defective sperm function and its association with the genesis of reactive oxygen species by human spermatozoa. J Reprod Fertil. 1987;81:459–69.CrossRefPubMed
43.
go back to reference Henkel R, Kierspel E, Stalf T, Mehnert C, Menkveld R, Tinneberg H-R, et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril. 2005;83:635–42.CrossRefPubMed Henkel R, Kierspel E, Stalf T, Mehnert C, Menkveld R, Tinneberg H-R, et al. Effect of reactive oxygen species produced by spermatozoa and leukocytes on sperm functions in non-leukocytospermic patients. Fertil Steril. 2005;83:635–42.CrossRefPubMed
44.
go back to reference Agarwal A, Makker K, Sharma R. Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol N Y N 1989. 2008;59:2–11. Agarwal A, Makker K, Sharma R. Clinical relevance of oxidative stress in male factor infertility: an update. Am J Reprod Immunol N Y N 1989. 2008;59:2–11.
45.
go back to reference Liu QY, Ribecco M, Pandey S, Walker PR, Sikorska M. Apoptosis-related functional features of the DNaseI-like family of nucleases. Ann N Y Acad Sci. 1999;887:60–76.CrossRefPubMed Liu QY, Ribecco M, Pandey S, Walker PR, Sikorska M. Apoptosis-related functional features of the DNaseI-like family of nucleases. Ann N Y Acad Sci. 1999;887:60–76.CrossRefPubMed
46.
go back to reference Oliveri M, Daga A, Cantoni C, Lunardi C, Millo R, Puccetti A. DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis. Eur J Immunol. 2001;31:743–51.CrossRefPubMed Oliveri M, Daga A, Cantoni C, Lunardi C, Millo R, Puccetti A. DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis. Eur J Immunol. 2001;31:743–51.CrossRefPubMed
47.
go back to reference Alghamdi AS, Funnell BJ, Bird SL, Lamb GC, Rendahl AK, Taube PC, et al. Comparative studies on bull and stallion seminal DNase activity and interaction with semen extender and spermatozoa. Anim Reprod Sci. 2010;121:249–58.CrossRefPubMed Alghamdi AS, Funnell BJ, Bird SL, Lamb GC, Rendahl AK, Taube PC, et al. Comparative studies on bull and stallion seminal DNase activity and interaction with semen extender and spermatozoa. Anim Reprod Sci. 2010;121:249–58.CrossRefPubMed
48.
go back to reference Ding Z, Qu F, Guo W, Ying X, Wu M, Zhang Y. Identification of sperm forward motility-related proteins in human seminal plasma. Mol Reprod Dev. 2007;74:1124–31.CrossRefPubMed Ding Z, Qu F, Guo W, Ying X, Wu M, Zhang Y. Identification of sperm forward motility-related proteins in human seminal plasma. Mol Reprod Dev. 2007;74:1124–31.CrossRefPubMed
Metadata
Title
Alterations in the proliferative/apoptotic equilibrium in semen of adolescents with varicocele
Authors
Larissa Berloffa Belardin
Paula Toni Del Giudice
Mariana Camargo
Paula Intasqui
Mariana Pereira Antoniassi
Ricardo Pimenta Bertolla
Agnaldo Pereira Cedenho
Publication date
01-12-2016
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 12/2016
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-016-0808-z

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