ORIGINAL ARTICLE
Cytochrome P450-2D6*4 polymorphism seminal relationship in infertile men A. Zalata1, A. Z. El-Samanoudy1, G. Osman1, S. Elhanbly2, H. A. Nada1 & T. Mostafa3 1 Medical Biochemistry Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 2 Dermatology & Andrology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 3 Andrology & Sexology Department, Faculty of Medicine, Cairo University, Cairo, Egypt
Keywords Acrosin activity—cytochrome 450—male infertility—polymorphism—semen Correspondence Dr Taymour Mostafa, MD, Andrology & Sexology Department, Faculty of Medicine, Cairo University, Cairo 11562, Egypt. Tel.: +2 01005150297; Fax: +2 23654133; E-mail: [email protected] Accepted: April 8, 2014 doi: 10.1111/and.12298
Summary This study aimed to assess cytochrome (CY) P450-2D6*4 polymorphism relationship with semen variables in infertile men. In all, 308 men were included; fertile normozoospermia (N) (n = 77), asthenozoospermia (A) (n = 70), asthenoteratozoospermia (AT) (n = 75) and oligoasthenoteratozoospermia (OAT) (n = 86). They were subjected to history taking, clinical examination, semen analysis, sperm acrosin activity, seminal malondialdehyde (MDA) and CYP4502D6*4 genotyping. CYP450-2D6*4 wild-type allele was represented in 76.5% of N, 70% of A, 66.7% of AT and 57.7% of OAT men where homozygous gene mutation was present in 5.9% of N, 20% of A, 26.6% of AT and 26.9% of OAT men, respectively. Sperm acrosin activity, sperm concentration, sperm motility, linear sperm velocity and sperm normal forms were significantly higher, and seminal MDA level was significantly lower in men with CYP4502D6*4 wild-type allele compared with men with homozygous mutation. It is concluded that CYP450-2D6*4 wild-type allele has higher frequency where homozygous-type allele has lower frequency in N men compared with A, AT and OAT men. Sperm acrosin activity index, sperm concentration, sperm motility, linear sperm velocity and sperm normal forms were significantly higher, and seminal MDA level was significantly lower in men with CYP4502D6*4 wild-type allele compared with men with homozygous mutation.
Introduction Genetic factors including gene mutations are lately demonstrated to be responsible for some cases of male infertility (Zalata et al., 2008, 2012a,b). Although several external factors were claimed to affect male reproduction, evidence that genetic factors play an important role is accumulating where reduced sperm count and fertility were associated with increased rate of chromosomal abnormalities (Olesen et al., 2001; Zalata et al., 2013a,b). Aydos et al. (2009) pointed that genetic polymorphisms of xenobiotic-metabolising enzymes could play an important role in infertility. Cytochrome P450 (CYP450) describes members of hemoprotein superfamily that catalyses the oxidative metabolism of endogenous substrates as steroids, fatty acids, prostaglandins, leukotrienes and biogenic amines (Lardone et al., 2010). CYP450s are found in the endoplasmic reticulum or mitochondrial fractions of many tissues and function either in catabolic pathways or in the © 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
biosynthesis of physiological molecules (Neve & Ingelman-Sundberg, 2010). CYP2D6 gene locus is highly polymorphic with more than 80 allelic variants. As a consequence, its activity ranges widely in a population comprising ultra-rapid, extensive, intermediate and poor metabolised phenotypes. CYP2D6 locus is on human chromosome 22q13.1 of DNA from somatic cell hybrid and is present in liver, peripheral blood leucocytes and other tissues (Gaedigk et al., 2005). Schur et al. (2001) pointed that in humans, CYP2D6*4 appears as two bands, at 105 and 250 bp, referred as wild-type allele determination. The appearance of an undigested 355-bp fragment indicates the presence of mutation of CYP2D6*4 gene, termed homozygous mutation. This explained that in normal CYP2D6 gene, there is a restriction site (-CCNGG-) at the boundary that is lost in individuals with G to A transition. As a consequence, these enzymes will not cut the genomic DNA in homozygous mutation. A mixture of the three fragments 1
CYP450 polymorphism
is indicative of the heterozygous mutation (Spurr et al., 1991). This study aimed to assess CYP2D6*4 gene polymorphism relationship with semen variables in infertile men. Materials and methods In all, 308 Egyptian men (mostly Caucasians) were recruited from the University Hospital after IRB approval and informed consents. These subjects were selected out of a total of 378 men during the period (January 2011– September 2013). They were grouped into healthy fertile men (N) (n = 77) [normozoospermic semen analysis and fathered a child within the last 2 years], asthenozoospermia (A) (n = 70) [infertile men with sperm count >15 million ml1, sperm motility 4%], asthenoteratozoospermia (AT) (n = 75) [infertile men with sperm count >15 million ml1, sperm motility 0.05 63.0 (49.3–84.8) P > 0.05 8.0 (1.15–64.0) P > 0.05 4.71 (1.18–10.85) P > 0.05 7.11 (2.11–43.89) P > 0.05
35.0 (27.3–43.7)
71.0 (61.2–74.7)
0.001 61.1 16.5a
index % P
GA (n = 38)
0.001 17.3 11.3a
% P
GG (n = 210)
0.001 1.3 0.7a
morphology
9.30 (4.26–11.09)
% P
0.001
0.001
0.001
N, normozoospermia; A, asthenozoospermia; AT, asthenoteratozoospermia; OAT, oligoasthenoteratozoospermia; GG, wild-type allele; GA, heterozygous allele; AA, homozygous allele. a Significant difference compared with N group.
© 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
34.9 (18.6–54.3)a P = 0.0191 19.1 (13.4–31.9)a P = 0.0093 61.6 (52.9–83.2) P > 0.05 6.0 (2–34.33)a P = 0.025 1.98 (0.72–7.01)a P = 0.002 13.51 (4.41–79.9)a P = 0.046
GG, wild-type allele; GA, heterozygous allele; AA, homozygous allele. a Significant difference compared with GG.
3
CYP450 polymorphism
Fig. 1 Agarose gel electrophoresis for restriction analysis PCR products of CYP450-2D6*4; two bands (250 bp, 105 bp, wild allele), one band (355 bp, homozygous allele) and 3 bands (355, 250 and 105 bp, heterozygous allele).
Discussion The current study demonstrated different genotypes of CYP2D6*4 among infertile men where there was a tendency to increase the frequency of wild-type gene in fertile normozoospermic men and increased frequency of homozygous mutations in OAT group. There was significantly lower acrosin activity index, sperm motility, sperm velocity, linear sperm velocity and sperm normal morphology in men with homozygous mutation of CYP2D6*4 compared with men with wild-type allele. Fritsche et al. (1998) investigated CYP1A1 polymorphisms in infertile Caucasian men showing increased frequencies of the Mspl polymorphism in the 30 -flanking region of CYP1A1 gene and a mutation in exon 7 causing an isoleucine–valine exchange in the heme-binding region of the enzyme. Liu et al. (2005) reported that CYP17, in addition to its role in steroidogenesis and androgen formation, is present in germ cells being essential for sperm function. Liu et al. (2005) added that deletion of one allele prevents genetic transmission causing male infertility followed by change in sexual behaviour due to androgen imbalance. Vani et al. (2009) showed a drastic decrease in the sperm count, sperm motility and increased dead spermaozoa in CC genotype of CYP1A1*2A compared with other genotypes in infertile Indian men. 4
A. Zalata et al.
Lambard et al. (2003) confirmed the presence of CYP450 aromatase (CYP450arom) transcript, a key enzyme responsible for the irreversible transformation of androgens into estrogens, in all normozoospermic purified samples. In men with CYP450 arom deficiency, Rochira et al. (2005) reported decreased sperm motility. In addition, the amount of P450 arom transcripts was found to be 30% lower in immotile versus motile sperms of healthy fertile men and a twofold decrease in spermatozoa from infertile men (Lambard et al., 2003; Jedrzejczak et al., 2006). Galeraud-Denis et al. (2008) added that spermatozoa of asthenospermic, teratospermic and asthenoteratospermic patients have 44%, 52% and 67% decreases in the amount of CYP450 arom transcripts compared with fertile controls. In addition, affected seminal variables could be explained on the light of the significant increase in reactive oxygen species (ROS) represented in elevated seminal MDA in homozygous mutation of CYP2D6*4 compared with men with wild-type allele. Nebert et al. (1989) reported that microsomal CYP2D6 produces ROS metabolites that could exert deleterious effects on sperm membrane composition and function. Yu et al. (2003) described CYP2D6 activity towards endogenous substrates producing ROS intermediates that causes injury to the cells. In their studies, Zalata et al. (1998), Taha et al. (2012) and Tawadrous et al. (2013) highlighted that seminal oxidative stress exerts a significant effect upon the functional quality of spermatozoa changing sperm membrane phospholipid composition by lowering its polyunsaturated fatty acids. In turn, these changes lower the membrane fluidity, reduce sperm acrosome reactivity and affect sperm fusogenic capacity (Depuydt et al., 1998; Mostafa et al., 2012). Points of limitation are the relatively small number of studied cases, the incomplete restriction enzyme digestion and the reported frequencies that were out of Hardy– Weinberg equilibrium. This indicates that more participants should be compared with a control population of the same ethnic race to explore the reasons for the difference, if any. It is concluded that CYP-2D6*4 wild-type allele has higher frequency where homozygous-type allele has lower frequency in N men compared with A, AT and OAT men. Sperm acrosin activity index, sperm motility and linear sperm velocity were significantly higher, and seminal MDA level was significantly lower in men with wildtype allele compared with men with homozygous mutation. Conflict of interest None. © 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
A. Zalata et al.
CYP450 polymorphism
References
(1989) The P450 superfamily: update listing of all genes and recommended nomenclature for the chromosomal loci. DNA 1:1–13. Neve EP, Ingelman-Sundberg M (2010) Cytochrome P450 proteins: retention and distribution from the endoplasmic reticulum. Curr Opin Drug Discov Devel 13:78–85. Olesen C, Hansen C, Bendsen E, Byskov A, Schwinger E, Pajares IL, Jensen PKA, Assche EV, Wahlstroem J, Lespinasse J, Tommerup N (2001) Identification of human candidate genes for male infertility. Mol Hum Reprod 7:11– 20. Rochira V, Granata AR, Madeo B, Zirilli L, Rossi G, Carani C (2005) Estrogens in males: what we have learned in last 10 years. Asian J Androl 7:3–20. Schur BC, Bjereke J, Nuwayhid N, Wong SH (2001) Genotyping of cytochrome P450-2D6*4 and *3 mutations using conventional PCR. Clin Chim Acta 308:25–31. Spurr NK, Alan C, Gough CA, Smith D, Wolf RC (1991) Genetic analysis of cytochrome P450 system. Methods Enzymol 206:149–166. Taha EA, Ez-Aldin AM, Sayed SK, Ghandour NM, Mostafa T (2012) Effect of smoking on sperm vitality, DNA integrity, seminal oxidative stress, zinc in fertile men. Urology 80:822– 825. Tawadrous GA, Aziz AA, Mostafa T (2013) Seminal soluble fas relationship with oxidative stress in infertile men with varicocele. Urology 82:820–823. Vani GT, Mukesh N, Siva Prasad B, Rama Devi P, Hema Prasad M, Usha Rani P, Pardhanandana Reddy P (2009) Association of CYP1A1*2A polymorphism with male infertility in Indian population. Clin Chim Acta 410:43–47. World Health Organization (2010) WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th edn. WHO press, Geneva, Switzerland. Yu AM, Idle JR, Herraiz T, Kupfer A, Gonzalez FJ (2003) Screening for endogenous substrates reveals that CYP2D6 is a 5-ethoxyindolethylamine O-demethylase. Pharmacogenetics 13:307–319. Zalata AA, Christophe AB, Depuydt CE, Schoonjans F, Comhaire FH (1998) The fatty acid composition of phospholipids of spermatozoa from infertile patients. Mol Hum Reprod 4:111–118. Zalata AA, Hassan AH, Nada HA, Bragais FM, Agarwal A, Mostafa T (2008) Follicle-stimulating hormone receptor polymorphism and seminal anti-M€ ullerian hormone in fertile and infertile men. Andrologia 40:392–397. Zalata AA, Morsy HK, Badawy A-N, Elhanbly S, Mostafa T (2012a) ACE gene insertion/deletion polymorphism seminal associations in infertile men. J Urol 187:1776–1780. Zalata A, El-Samanoudy AZ, Shaalan D, El-Baiomy Y, Taymour M, Mostafa T (2012b) Seminal clusterin gene expression associated with seminal variables in fertile and infertile men. J Urol 188:1260–1264. Zalata AA, Mokhtar N, Badawy A-N, Othman G, Alghobary M, Mostafa T (2013a) Androgen receptor expression
Aydos SE, Taspinar M, Sunguroglu A, Aydos K (2009) Association of CYP1A1 and glutathione S-transferase polymorphisms with male factor infertility. Fertil Steril 92:541–547. Depuydt C, Zalata A, Christophe A, Comhaire FH (1998) Mechanisms of sperm deficiency in male accessory gland infection. Andrologia 30:29–33. Draper HH, Squires EJ, Mahmoodi H, Wu J, Agarwal S, Hadley MA (1993) A comparative evaluation of thiobarbituric acid methods for the determination of malondialdehydes in biological materials. Free Radic Biol Med 15:353–363. Fritsche E, Schuppe HC, Dohr O, Ruzicka T, Gleichmann E, Abel J (1998) Increased frequencies of cytochrome P4501A1 polymorphisms in infertile men. Andrologia 30:125–128. Gaedigk A, Bhathena A, Ndjountche L, Pearce RE, AbdelRahman SM, Alander SW, Bradford LD, Leeder JS (2005) Identification and characterization of novel sequence variations in the cytochrome P4502D6 (CYP2D6) gene in African Americans. Pharmacogenomics J 5:173–182. Galeraud-Denis I, de Vienne C, Said L, Chocat A, Carreau S (2008) Differential expression of aromatase transcript and protein in human spermatozoa from normozoospermic and teratozoospermic patients. Int J Androl 31:1. Henkel R, Muller C, Miska W, Schill WB, Kleinstein J, Gips H (1995) Acrosin activity of human spermatozoa by means of a simple gelatinolytic technique: a method useful for IVF. J Androl 16:272–277. Jedrzejczak P, Januchowski R, Taszarek-Hauke G, Laddach R, Pawelczyk L, Jagodzinski PP (2006) Quantitative analysis of CCR5 chemokine receptor and cytochrome P450 aromatase transcripts in swim-up spermatozoa isolated from fertile and infertile men. Arch Androl 52:335–341. Lambard S, Galeraud-Denis I, Boura€ıma H, Bourguiba S, Chocat A, Carreau S (2003) Expression of aromatase in human ejaculated spermatozoa: a putative marker of motility. Mol Hum Reprod 9:117–124. Lardone MC, Castillo P, Valdevenito R, Ebensperger M, Ronco AM, Pommer R, Piottante A, Castro A (2010) P450aromatase activity and expression in human testicular tissues with severe spermatogenic failure. Int J Androl 33:650–660. Liu Y, Yao ZX, Bendavid C, Borgmeyer C, Han Z, Cavalli LR, Chan WY, Folmer J, Zirkin BR, Haddad BR, Gallicano GI, Papadopoulos V (2005) Haplo insufficiency of cytochrome P450 17alpha-hydroxylase/17,20 lyase (CYP17) causes infertility in male mice. Mol Endocrinol 19:2380–2389. Mostafa T, Anis T, El Nashar A, Imam H, Osman I (2012) Seminal plasma reactive oxygen species-antioxidants relationship with varicocele grade. Andrologia 44:66–69. Nebert DW, Nelson DR, Adesnik M, Coon MJ, Estabrook RW, Gonzalez FJ, Guengerich FP, Gunsalus IC, Johnson EF, Kemper B, Levin W, Phillips IR, Sato R, Waterman MR
© 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
5
CYP450 polymorphism
relationship with semen variables in infertile men with varicocele. J Urol 189:2243–2247. Zalata A, Atwa A, El-Naser BA, Aziz A, El-Baz R, Elhanbly S, Mostafa T (2013b) Tumor necrosis factor-a gene
6
A. Zalata et al.
polymorphism relationship to seminal variables in infertile men. Urology 81:962–966.
© 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
Cytochrome P450-2D6*4 polymorphism seminal relationship in infertile men A. Zalata1, A. Z. El-Samanoudy1, G. Osman1, S. Elhanbly2, H. A. Nada1 & T. Mostafa3 1 Medical Biochemistry Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 2 Dermatology & Andrology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 3 Andrology & Sexology Department, Faculty of Medicine, Cairo University, Cairo, Egypt
Keywords Acrosin activity—cytochrome 450—male infertility—polymorphism—semen Correspondence Dr Taymour Mostafa, MD, Andrology & Sexology Department, Faculty of Medicine, Cairo University, Cairo 11562, Egypt. Tel.: +2 01005150297; Fax: +2 23654133; E-mail: [email protected] Accepted: April 8, 2014 doi: 10.1111/and.12298
Summary This study aimed to assess cytochrome (CY) P450-2D6*4 polymorphism relationship with semen variables in infertile men. In all, 308 men were included; fertile normozoospermia (N) (n = 77), asthenozoospermia (A) (n = 70), asthenoteratozoospermia (AT) (n = 75) and oligoasthenoteratozoospermia (OAT) (n = 86). They were subjected to history taking, clinical examination, semen analysis, sperm acrosin activity, seminal malondialdehyde (MDA) and CYP4502D6*4 genotyping. CYP450-2D6*4 wild-type allele was represented in 76.5% of N, 70% of A, 66.7% of AT and 57.7% of OAT men where homozygous gene mutation was present in 5.9% of N, 20% of A, 26.6% of AT and 26.9% of OAT men, respectively. Sperm acrosin activity, sperm concentration, sperm motility, linear sperm velocity and sperm normal forms were significantly higher, and seminal MDA level was significantly lower in men with CYP4502D6*4 wild-type allele compared with men with homozygous mutation. It is concluded that CYP450-2D6*4 wild-type allele has higher frequency where homozygous-type allele has lower frequency in N men compared with A, AT and OAT men. Sperm acrosin activity index, sperm concentration, sperm motility, linear sperm velocity and sperm normal forms were significantly higher, and seminal MDA level was significantly lower in men with CYP4502D6*4 wild-type allele compared with men with homozygous mutation.
Introduction Genetic factors including gene mutations are lately demonstrated to be responsible for some cases of male infertility (Zalata et al., 2008, 2012a,b). Although several external factors were claimed to affect male reproduction, evidence that genetic factors play an important role is accumulating where reduced sperm count and fertility were associated with increased rate of chromosomal abnormalities (Olesen et al., 2001; Zalata et al., 2013a,b). Aydos et al. (2009) pointed that genetic polymorphisms of xenobiotic-metabolising enzymes could play an important role in infertility. Cytochrome P450 (CYP450) describes members of hemoprotein superfamily that catalyses the oxidative metabolism of endogenous substrates as steroids, fatty acids, prostaglandins, leukotrienes and biogenic amines (Lardone et al., 2010). CYP450s are found in the endoplasmic reticulum or mitochondrial fractions of many tissues and function either in catabolic pathways or in the © 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
biosynthesis of physiological molecules (Neve & Ingelman-Sundberg, 2010). CYP2D6 gene locus is highly polymorphic with more than 80 allelic variants. As a consequence, its activity ranges widely in a population comprising ultra-rapid, extensive, intermediate and poor metabolised phenotypes. CYP2D6 locus is on human chromosome 22q13.1 of DNA from somatic cell hybrid and is present in liver, peripheral blood leucocytes and other tissues (Gaedigk et al., 2005). Schur et al. (2001) pointed that in humans, CYP2D6*4 appears as two bands, at 105 and 250 bp, referred as wild-type allele determination. The appearance of an undigested 355-bp fragment indicates the presence of mutation of CYP2D6*4 gene, termed homozygous mutation. This explained that in normal CYP2D6 gene, there is a restriction site (-CCNGG-) at the boundary that is lost in individuals with G to A transition. As a consequence, these enzymes will not cut the genomic DNA in homozygous mutation. A mixture of the three fragments 1
CYP450 polymorphism
is indicative of the heterozygous mutation (Spurr et al., 1991). This study aimed to assess CYP2D6*4 gene polymorphism relationship with semen variables in infertile men. Materials and methods In all, 308 Egyptian men (mostly Caucasians) were recruited from the University Hospital after IRB approval and informed consents. These subjects were selected out of a total of 378 men during the period (January 2011– September 2013). They were grouped into healthy fertile men (N) (n = 77) [normozoospermic semen analysis and fathered a child within the last 2 years], asthenozoospermia (A) (n = 70) [infertile men with sperm count >15 million ml1, sperm motility 4%], asthenoteratozoospermia (AT) (n = 75) [infertile men with sperm count >15 million ml1, sperm motility 0.05 63.0 (49.3–84.8) P > 0.05 8.0 (1.15–64.0) P > 0.05 4.71 (1.18–10.85) P > 0.05 7.11 (2.11–43.89) P > 0.05
35.0 (27.3–43.7)
71.0 (61.2–74.7)
0.001 61.1 16.5a
index % P
GA (n = 38)
0.001 17.3 11.3a
% P
GG (n = 210)
0.001 1.3 0.7a
morphology
9.30 (4.26–11.09)
% P
0.001
0.001
0.001
N, normozoospermia; A, asthenozoospermia; AT, asthenoteratozoospermia; OAT, oligoasthenoteratozoospermia; GG, wild-type allele; GA, heterozygous allele; AA, homozygous allele. a Significant difference compared with N group.
© 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
34.9 (18.6–54.3)a P = 0.0191 19.1 (13.4–31.9)a P = 0.0093 61.6 (52.9–83.2) P > 0.05 6.0 (2–34.33)a P = 0.025 1.98 (0.72–7.01)a P = 0.002 13.51 (4.41–79.9)a P = 0.046
GG, wild-type allele; GA, heterozygous allele; AA, homozygous allele. a Significant difference compared with GG.
3
CYP450 polymorphism
Fig. 1 Agarose gel electrophoresis for restriction analysis PCR products of CYP450-2D6*4; two bands (250 bp, 105 bp, wild allele), one band (355 bp, homozygous allele) and 3 bands (355, 250 and 105 bp, heterozygous allele).
Discussion The current study demonstrated different genotypes of CYP2D6*4 among infertile men where there was a tendency to increase the frequency of wild-type gene in fertile normozoospermic men and increased frequency of homozygous mutations in OAT group. There was significantly lower acrosin activity index, sperm motility, sperm velocity, linear sperm velocity and sperm normal morphology in men with homozygous mutation of CYP2D6*4 compared with men with wild-type allele. Fritsche et al. (1998) investigated CYP1A1 polymorphisms in infertile Caucasian men showing increased frequencies of the Mspl polymorphism in the 30 -flanking region of CYP1A1 gene and a mutation in exon 7 causing an isoleucine–valine exchange in the heme-binding region of the enzyme. Liu et al. (2005) reported that CYP17, in addition to its role in steroidogenesis and androgen formation, is present in germ cells being essential for sperm function. Liu et al. (2005) added that deletion of one allele prevents genetic transmission causing male infertility followed by change in sexual behaviour due to androgen imbalance. Vani et al. (2009) showed a drastic decrease in the sperm count, sperm motility and increased dead spermaozoa in CC genotype of CYP1A1*2A compared with other genotypes in infertile Indian men. 4
A. Zalata et al.
Lambard et al. (2003) confirmed the presence of CYP450 aromatase (CYP450arom) transcript, a key enzyme responsible for the irreversible transformation of androgens into estrogens, in all normozoospermic purified samples. In men with CYP450 arom deficiency, Rochira et al. (2005) reported decreased sperm motility. In addition, the amount of P450 arom transcripts was found to be 30% lower in immotile versus motile sperms of healthy fertile men and a twofold decrease in spermatozoa from infertile men (Lambard et al., 2003; Jedrzejczak et al., 2006). Galeraud-Denis et al. (2008) added that spermatozoa of asthenospermic, teratospermic and asthenoteratospermic patients have 44%, 52% and 67% decreases in the amount of CYP450 arom transcripts compared with fertile controls. In addition, affected seminal variables could be explained on the light of the significant increase in reactive oxygen species (ROS) represented in elevated seminal MDA in homozygous mutation of CYP2D6*4 compared with men with wild-type allele. Nebert et al. (1989) reported that microsomal CYP2D6 produces ROS metabolites that could exert deleterious effects on sperm membrane composition and function. Yu et al. (2003) described CYP2D6 activity towards endogenous substrates producing ROS intermediates that causes injury to the cells. In their studies, Zalata et al. (1998), Taha et al. (2012) and Tawadrous et al. (2013) highlighted that seminal oxidative stress exerts a significant effect upon the functional quality of spermatozoa changing sperm membrane phospholipid composition by lowering its polyunsaturated fatty acids. In turn, these changes lower the membrane fluidity, reduce sperm acrosome reactivity and affect sperm fusogenic capacity (Depuydt et al., 1998; Mostafa et al., 2012). Points of limitation are the relatively small number of studied cases, the incomplete restriction enzyme digestion and the reported frequencies that were out of Hardy– Weinberg equilibrium. This indicates that more participants should be compared with a control population of the same ethnic race to explore the reasons for the difference, if any. It is concluded that CYP-2D6*4 wild-type allele has higher frequency where homozygous-type allele has lower frequency in N men compared with A, AT and OAT men. Sperm acrosin activity index, sperm motility and linear sperm velocity were significantly higher, and seminal MDA level was significantly lower in men with wildtype allele compared with men with homozygous mutation. Conflict of interest None. © 2014 Blackwell Verlag GmbH Andrologia 2014, xx, 1–6
A. Zalata et al.
CYP450 polymorphism
References
(1989) The P450 superfamily: update listing of all genes and recommended nomenclature for the chromosomal loci. DNA 1:1–13. Neve EP, Ingelman-Sundberg M (2010) Cytochrome P450 proteins: retention and distribution from the endoplasmic reticulum. Curr Opin Drug Discov Devel 13:78–85. Olesen C, Hansen C, Bendsen E, Byskov A, Schwinger E, Pajares IL, Jensen PKA, Assche EV, Wahlstroem J, Lespinasse J, Tommerup N (2001) Identification of human candidate genes for male infertility. Mol Hum Reprod 7:11– 20. Rochira V, Granata AR, Madeo B, Zirilli L, Rossi G, Carani C (2005) Estrogens in males: what we have learned in last 10 years. Asian J Androl 7:3–20. Schur BC, Bjereke J, Nuwayhid N, Wong SH (2001) Genotyping of cytochrome P450-2D6*4 and *3 mutations using conventional PCR. Clin Chim Acta 308:25–31. Spurr NK, Alan C, Gough CA, Smith D, Wolf RC (1991) Genetic analysis of cytochrome P450 system. Methods Enzymol 206:149–166. Taha EA, Ez-Aldin AM, Sayed SK, Ghandour NM, Mostafa T (2012) Effect of smoking on sperm vitality, DNA integrity, seminal oxidative stress, zinc in fertile men. Urology 80:822– 825. Tawadrous GA, Aziz AA, Mostafa T (2013) Seminal soluble fas relationship with oxidative stress in infertile men with varicocele. Urology 82:820–823. Vani GT, Mukesh N, Siva Prasad B, Rama Devi P, Hema Prasad M, Usha Rani P, Pardhanandana Reddy P (2009) Association of CYP1A1*2A polymorphism with male infertility in Indian population. Clin Chim Acta 410:43–47. World Health Organization (2010) WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th edn. WHO press, Geneva, Switzerland. Yu AM, Idle JR, Herraiz T, Kupfer A, Gonzalez FJ (2003) Screening for endogenous substrates reveals that CYP2D6 is a 5-ethoxyindolethylamine O-demethylase. Pharmacogenetics 13:307–319. Zalata AA, Christophe AB, Depuydt CE, Schoonjans F, Comhaire FH (1998) The fatty acid composition of phospholipids of spermatozoa from infertile patients. Mol Hum Reprod 4:111–118. Zalata AA, Hassan AH, Nada HA, Bragais FM, Agarwal A, Mostafa T (2008) Follicle-stimulating hormone receptor polymorphism and seminal anti-M€ ullerian hormone in fertile and infertile men. Andrologia 40:392–397. Zalata AA, Morsy HK, Badawy A-N, Elhanbly S, Mostafa T (2012a) ACE gene insertion/deletion polymorphism seminal associations in infertile men. J Urol 187:1776–1780. Zalata A, El-Samanoudy AZ, Shaalan D, El-Baiomy Y, Taymour M, Mostafa T (2012b) Seminal clusterin gene expression associated with seminal variables in fertile and infertile men. J Urol 188:1260–1264. Zalata AA, Mokhtar N, Badawy A-N, Othman G, Alghobary M, Mostafa T (2013a) Androgen receptor expression
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