The impact of seminal zinc and fructose concentration on human sperm characteristic

This study assessed the association between fructose and zinc concentration and various seminal characteristics. Fructose and zinc in semen reflect the secretory function of seminal vesicles. These tests may help in assessing the diagnosis and the management of male infertility

JOURNAL OF MEDICAL RESEARCH THE IMPACT OF SEMINAL ZINC AND FRUCTOSE CONCENTRATION ON HUMAN SPERM CHARACTERISTIC Vu Thi Huyen, Nguyen Thi Trang, Luong Thi Lan Anh, Vu To Giang, Bui Bich Mai, Nguyen Xuan Tung Department of Biomedical and Genetics, Hanoi Medical University, Hanoi, Vietnam This study assessed the association between fructose and zinc concentration and various seminal characteristics Fructose and zinc in semen reflect the secretory function of seminal vesicles These tests may help in assessing the diagnosis and the management of male infertility Seminal plasma was gathered from 180 males who averaged 31.1 ± 3,6 in age A specific complexant was used to form a stable coloured complex with fructose or zinc The colour intensity of the complex in a determining wavelength is proportional to the amount of fructose or zinc present in the sample The study found that seminal fructose concentration was significantly lower in the oligozoospermic group and the azoospermic group in comparison to the with normozoospermic group There were also many significant differences in zinc’s concentration in semen when two of three groups were compared with one another In conclusion, the role of seminal fructose concentration lie not only in the assessing seminal vesicle dysfunction, but also, in conjunction with other seminal properties could give a useful indication of male reproductive function, whilst seminal zinc concentration might not be most appropriate for the assessment of male reproductive dysfunction Keywords: infertility, seminal fructose, seminal zinc, azoospermia I INTRODUCTION As many reasons cause male infertility, it is essential to identify appropriate methods to diagnose the underlying cause many tests have been applied previously, such as semen analysis, genetic tests and hormone methods Recently, some of these biochemical markers zinc and fructose, are like increasingly recognized as important for Corresponding author: Nguyen Thi Trang, Department of Biomedical and Genetics,Hanoi Medical University Email: trangnguyen@hmu.edu.vn Received: 03 June 2017 Accepted: 16 November 2017 JMR 111 E2 (2) - 2018 diagnosing the cause of male infertility Fructose is essential for spermatozoa metabolism and motility Fructose is an energy source of spermatozoa It is produced by the seminal vesicles with some contribution from the ampulla of the ductus deferens [1; 2] Absence of fructose in semen is indicative of ejaculatory duct obstruction or seminal vesicle dysfunction [3; 4] Apart from fructose, zinc is another factor that is essential for the male reproductive system Deficiency of zinc in the reproductive system causes hypogonadism and gonadal 17 JOURNAL OF MEDICAL RESEARCH hypofunction [5; 6] Many studies have shown that zinc plays an important role in sperm mobility an the normal development of the testicles and prostate [2; 7; 8] However, in Vietnam, knowledge about the relationship between seminal zinc and fructose concentration in human sperm is scare Therefore, the purpose of this study was to determine the association between fructose and zinc concentration and various seminal characteristics in men II SUBJECTS AND METHODS Subjects The study design was descriptive Fructose and zinc concentration was measured in the seminal plasma of 180 patients, who visited the Fertility Department of Hanoi Medical University Hospital from March, 2016 to March, 2017 after semen analysis tests showing abnormal seminal characteristics (sperm concentration, total count, motility, progressive motility) All the samples were analyzed according to the World Health Organization criteria (1992) On the basis of the assessed parameters, sperm concentration and sperm motility were considered as the most important parameters Method Measuring the concentration of fructose and zinc After semen analysis, samples were centrifuged at 1500 x g for 10 and zinc and fructose concentrations assayed from the supernatant (i.e seminal plasma) Zinc concentration was assessed using spectrophotometry (5- Br- PAPS 18 method) – direct colorimetric test without deproteinization of the sample At pH 8.6, in a buffered media, zinc react with specific complexant 5-Br-PAPS form a stable color compound Fructose content in seminal plasma was determined by the resorcinol method where fructose reacts with resorcinol in concentrated hydrochloric acid (HCl) solution to form a red compound Measure the coloric complex of Zinc and Fructose at a wavelength of 560 nm against blanks (ROE, 1976) Statistical analysis Statistical analysis was performed using SPSS version 16.0 The means were compared using student t test The statistical tests were considered to be significant at the p ≤ 0.05 level Ethics Ethical approval to conduct the study was sought from the Hanoi Medical University Permission to use data from the Hanoi Medical University Hospital was sought from the hospital authority All the information from the database was kept under strict confidentiality No names were recorded III RESULTS Fructose concentration and seminal parameters Table shows that seminal fructose in oligozoospermia was significantly higher than normozoospermia (p < 0.05) Besides, the mean sperm concentration (133.808 ± 48.215 billion/ mL), and the mean vitality (86.483 ± 3.218 %) and the mean progressive motility (11.250 ± 10.157 %) in JMR 111 E2 (2) - 2018 JOURNAL OF MEDICAL RESEARCH males with normozoospermia were significantly higher than that in males with oligozoospermia (5.633 ± 4.992 billion/ mL and 58.183 ± 18.14 % and 11.250 ± 10.157 % respectively) (p < 0.01) Table Seminal fructose and some characteristics of the semen (Independent sample T – test) Normozoospermia Mean± SD Oligozoospermia Mean± SD p-value of fructose test Fructose 1.601 ± 0.604 1.881 ± 0.640 < 0.05 Sperm concentration (billion/ml) 133.808 ± 48.215 5.633 ± 4.992 < 0.01 Vitality (%) 86.483 ± 3.218 58.183 ± 18.114 < 0.01 Progressive motility (%) 54.667 ± 9.278 11.250 ± 10.157 < 0.01 Some sperm characteristics and seminal fructose concentration are shown in the graph below The results suggest is a significant correlation at the 0.05 level (2-tailed) between seminal fructose concentration and sperm progressive motility (z = -0.183; p < 0.05) (Spearman test) (Figure 1, Figure and Figure 3) Figure Correlation between seminal fructose concentration (g/l) and sperm concentration (billion/ml) JMR 111 E2 (2) - 2018 19 JOURNAL OF MEDICAL RESEARCH Figure Correlation between seminal fructose concentration (g/l) and sperm vitality (%) Figure Correlation between seminal fructose concentration (g/l) and sperm progressive motility (%) Zinc concentration and seminal parameters Table shows the following: • The progressive mobility of the low zinc concentration group was 16.87 ± 10.67%, lower than that of the normal zinc concentration group (49.93 ± 15.35%) This difference is statistically significant (z= -11.481, p < 0.01) 20 JMR 111 E2 (2) - 2018 JOURNAL OF MEDICAL RESEARCH • There was no statistically significant difference in mean non- progressive motility of males with low zinc concentration compared to males with normal zinc concentration (p= 0.19) Table Seminal zinc concentration and motility of the sperm (Mann – Whitney test) Low zinc concentration (n = 84) Normal zinc concentration (n = 96) z p-value Progressive motility (%) 16.87 ± 10.67 49.93 ± 15.35 - 11.481 < 0.01 Non- progressive motility (%) 3.64 ± 2.07 4.07 ± 4.63 - 1.301 > 0.05 Immotile (%) 73.00 ± 21.42 44.07 ± 15.43 10.433 < 0.01 The low zinc concentration group had an immotile percentage of 73.00 ± 21.42% was higher than the normal zinc concentration group (44.07 ± 15.43%) (z = 10.433) This difference was statistically significant with p < 0.001 Seminal zinc concentration showed a significant positive correlation (r = 0.596) with sperm progressive motility (p < 0.01) Negative correlations with sperm immotile (r = - 0.527) which were observed reached statistical significance (p < 0.01) Figure Correlation between seminal zinc concentration (g/l) and sperm progressive motility (%) (r = 0.596; p < 0.01) ( Spearman test) JMR 111 E2 (2) - 2018 21 JOURNAL OF MEDICAL RESEARCH Figure Correlation between seminal zinc concentration (g/l) and sperm immotile (%) (r = - 0.527; p < 0.01) ( Spearman test) IV DISCUSSION Fructose is a main carbohydrate source in seminal plasma and necessary for sperm motility [9; 10] The measurement of seminal fructose has been used in many laboratories Therefore, the World Health Organization manual recommends measurement of seminal fructose as a marker of seminal vesicular function [11] Methods for determination of seminal fructose include gas chromatography, indole coloration, and resorcinol coloration In particular, the resorcinol method has been used widely in clinical andrology laboratories for its simplicity operational, and high specificity Fructose is the primary source of energy for all sperm activities The higher the sperm concentration, vitality, and, motility the lower fructose will be [2; 4] Lu 22 (2007) reported that when sperm motility increased, fructose decreased, and in vitro, sperm continued using fructose [4] Normal seminal fructose concentration confirms normal levels of testosterone and function of vesicles and vas deferens [12] Biswas et al (1978) also reported that when seminal fructose concentration decreased, sperm concentration and mobility increased [13] Furthermore, Lewis Jones et al.,1996 found that fructose concentrations were inversely ratio to sperm motility with R = - 0,062 (p < 0.05) [7] However, Andrade Rocha (2001) found contrary evidence that that seminal fructose concentration was related to sperm concentration, survival, motility and morphology, but the association was not statistically significant [14] In Amidu (2012), seminal fructose concentration negatively correlated with sperm motility JMR 111 E2 (2) - 2018 JOURNAL OF MEDICAL RESEARCH (R = - 0.04) but was also not statistically significant [15] Fructose concentration was inversely celated to sperm concentration (R = - 0.21) anh this correlation was significant at 0.05 [16] Determination of seminal fructose concentration has been used in the examination of obstructive azoospermia and inflammation of male accessory glands [11; 12; 15] Inflammation may lead to atrophy of the seminal vesicles and low seminal fructose concentration When ejaculatory ducts are blocked, fructose concentration in seminal plasma usually decreases and may become undetectable [12; 17] Additionally, determination of seminal plasma fructose concentration is useful for auxiliary diagnosis of obstructive and nonobstructive azoospermia Seminal fructose concentration in non-obstructive azoospermia is usually higher than or equal to that in males of normal fertility [9] However, the fructose concentration in seminal plasma of patients with obstructive azoospermia is usually absent or significantly lower than that in men of normal fertility [12; 15] Absence of seminal fructose has also been found in patients with congenital vas deferens-seminal vesicle developmental defect (Kise et al., 2000; Kumar et al., 2005) Therefore, our results are consistent with those reported studies in other international One of the biochemical processes related to genital fluid mixing is the regulation of the fraction of free seminal zinc, which can interact with spermatozoa Zinc is first secreted in prostatic fluid in forms available for sperm cells (free zinc and zinc-citrate complex) During ejaculation, JMR 111 E2 (2) - 2018 however, a partial redistribution of the ion from citrate to very high affinity vesicular ligands reduces the unbound zinc fraction [18 - 20] The measurement of zinc in human seminal plasma is important in the evaluation of male infertility In the present study, the level of zinc in seminal plasma was found to be mor frequently immotile in the zinc concentration group was higher (73.00 ± 21.42%) than in the group with normal zinc concentration (44.07 ± 15.43%) (z = 10.433) A positive correlation between zinc levels and sperm concentration and motility was also observed in our study This isin accordance with previous studies of Doshi et al., Hussain et al., Badade et al., Atig et al., and Abed [21 - 25] Eliasson and Lindholme et al., in contrast could not find any correlation between zinc concentration and sperm density, motility, or morphology [26] Fuse et al., found a positive correlation between zinc and sperm concentration and motility, but no correlation with sperm morphology was observed [27] Mankad et al., found a positive correlation between zinc and sperm count, but no significant correlation between zinc and sperm motility [28] Thus, it seems that zinc is important for semen quality The low zinc levels in infertile men in our study might be attributed to disorders in the prostate excretory function or possibly to asymptomatic prostate infection Omu (1998), Hadwan (2013), and others found that sperm motility increased after treatment with zinc supplementation [29 23 JOURNAL OF MEDICAL RESEARCH - 33] However, Omar F Abdul-Rasheed (2009) found no correlation between zinc concentrations in semen and sperm motility [34] V CONCLUSION The seminal fructose concentration of the normozopermia group is significantly lower than oligozoospermia group Fructose seminal concentration correclated with sperm motility The progressive motility in the low zinc concentration group is significantly lower than that of the normal zinc concentration group The number of immotile sperm in the low zinc concentration group is significantly higher than that of the normal zinc concentration group Zinc concentration has a positive correlation with sperm progressive motility and a negative correlation with immotile both are statistically significant Acknowledgements The authors would like to take this opportunity to extend their sincere thanks to the Ministry of Health for providing financial support for the study They also are grateful for the technical support form the Hanoi Medical University Hospital for assaying of seminal fructose and zinc REFERENCES Schoenfeld C, Amelar RD, Dubin L, Numeroff M (1979) Prolactin,fructose, and zinc levels found in human seminal plasma Fertil Steril 32(2) , 206 - 208 Biswas S., Ferguson K.M., Stedronska J et al (1978) Fructose and hormone levels in semen: their correlations 24 with sperm counts and motility Fertility and Sterility, 2, 200 - 204 Aumuller G, Riva A (1992) Morphology and functions of the human seminal vesicle Andrologia 24(4): 183 196 Lu C.J (2007) Standardization and quality control for determination of fructose in seminal plasma Journal of Andrology, 28 (2), 207 - 213 Sandstead HH, Prasad AS, Schulert AR, Farid Z, Miale A, Jr., Bassilly S, et al (1967) Human zinc deficiency, endocrine manifestations and response to treatment Am J Clin Nutr 20(5):422 - 442 Omu A.E, Dashti H, Al Othman S (1998) Treatment of asthenozoospermic with zinc sulphate: andrological , immunological and obstetric outcome Eur J Obstet Gynecol Reprod Biol, 79, 179 - 184 Lewis Jones D.I., Aird I.A., Biljan M.M (1996) Effects of sperm activity on zinc and fructose concentrations in seminal plasma Oxford Journals, Human Reproduction, 11 (11), 2465 – 2467 Basil Oied Mohammed Saleh, Nawal Khiry Hussain, Ali Yakub Majid et al (2008) Status of Zinc and Copper Concentrations in Seminal Plasma of Male Infertility and Their Correlation with Various Sperm Parameters The Iraqi postgraduate medical journal, 7, 76 - 80 Buckett WM, Lewis-Jones DI (2002) Fructose concentrations in seminal plasma from men with nonobstructive azoospermia Arch Androl; 48 , 23 – 27 10 Santiani A., Huanca W., Sapana R., Huanca T., Sepulveda N., Sanchez JMR 111 E2 (2) - 2018 JOURNAL OF MEDICAL RESEARCH R.(2005) Effects on the quality of frozenthawed alpaca (Lama pacos) semen using two different cryoprotectants and extenders Asian J Androl, , 303 – 309 11 World Health Organization (1999) Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction 4th ed Cambridge, United Kingdom: Cambridge University Press; – 10 12 WHO (1987) Laboratory manual for the examination of human semen and semen - cervical mucus interaction, Cambridge University Press 13 Biswas S., Ferguson K.M., Stedronska J et al (1978) Fructose and hormone levels in semen: their correlations with sperm counts and motility Fertil Steril, 2, 200 – 204 14 Andrade Rocha F.T (2001) Sperm parameters in men with suspected infertility Sperm characteristics, strict criteria sperm morphology analysis and hypoosmotic swelling test J Reprod Med., 46, 577 – 582 15 Amidu N., Owiredu W.K.B.A, Bekoe M.A.T (2012) The impact of seminal zinc and fructose concentration on human sperm characteristic Journal of Medical and Biomedical Sciences, (1), 14 – 20 16 Yassa D.A., Idriss W.K., Atassi M.E et al (2001) The diagnostic value of seminal α – glucosidase enzyme index for sperm motility and fertilizing capacity Saudi Medical Journal, 22 (11), 987 – 991 17 Coppens L (1997) Diagnosis and treatment of obstructive seminal vesicle pathology Acta Urol Belg, 65 (2): 11 - 19 18 Björndahl L, Kvist U (1990) JMR 111 E2 (2) - 2018 Influence of seminal vesicular fluid on the zinc content of human sperm chromatin Int J Androl, 13, 232 - 237 19 Arver S, Eliasson R (1982) Zinc and zinc ligands in human seminal plasma II contribution by ligands of different origin to the zinc binding properties of seminal plasma Acta Physiol Scand, 115, 217 - 224 20 Kvist U, Kjellberg S, Björndahl L, Soufir JC, Arver S.(1990), Seminal fluid from men with agenesis of the wolffian ducts: zincbinding properties and effects on sperm chromatin stability Int J Androl,13, 245 - 252 21 Doshi H, Heana O, Hemali T, Minal M, Sunil K (2008) Zinc levels in seminal plasma and its relationship with seminal characteristics Journal of Obstetrics and Gynecology of India, 58, 152 - 155 22 Hussain NK, Rzoqi SS, Numan AW, Ali DT (2011) A comparative study of fructose, zinc and copper levels in seminal plasma in fertile and infertile men Iraqi Journal of Medical Sciences (1):48 23 Badade ZG, More KM, Narshetty JG, Badade VZ, Yadav BK (2011) Human seminal oxidative stress: correlation with antioxidants and sperm quality parameters Annals of Biological Research, (5), 351 – 59 24 Atig F, Raffa M, Habib BA, Kerkeni A, Saad A, Ajina M(2012) Impact of seminal trace element and glutathione levels on semen quality of Tunisian infertile men BMC Urol, 12, 25 Abed AA (2013) Essence of some trace elements in seminal fluid and their role in infertility Int J Chem and Life Sciences, 02(6):1179–84 25 JOURNAL OF MEDICAL RESEARCH 26 Eliasson R, Lindholme C (1971) Zinc in human seminal plasma Andrologia, 39(4):147 – 153 27 Fuse H, Kazama T, Ohta S, Fujiuchi Y(1999) Relationship between zinc concentrations in seminal plasma and various sperm parameters Int Urol Nephrol, 31(3), 401 – 08 28 Mankad M, Sathawara NG, Doshi H, Saiyed HN, Kumar S (2006) Seminal plasma zinc concentration and á-glucosidase activity with respect to semen quality Biol Trace Elem Res, 110 (2), 97 – 106 29 Marmar J.L, Katz S, Praiss D.E et al (1975) Semen zinc levels in infertile and postvasectomy patients and patients with prostatitis Fertil Steril, 26,1057 – 1063 30 Hartoma T.R, Nahoul K, Netter A (1977) Zinc, plasma androgens and male 26 sterility Lancet, 2, 1125 – 1126 31 Mahajan S.K, Abbasi A.A, Prasad A.S et al (1982) Effect of oral zinc therapy on gonadal function in hemodialysis patients A double-blind study Ann Intern Med, 7, 357 – 361 32 Wong W.Y, Merkus H.M, Thomas C.M et al (2002) Effects of folic acid and zinc sulfate on male factor subfertility: a doubleblind, randomized, placebo-controlled trial Fertil Steril, 77, 491 – 498 33 Caldamone A.A, Freytag M.K, Cockett A.T (1979) Seminal zinc and male infertility Urology,13, 280 – 281 34 Omar F Abdul-Rasheed (2009) The relationship between seminal plasma zinc levels and high molecular weight zinc binding protein and sperm motility in Iraqi infertile men Saudi Med J, 30, 485 - 489 JMR 111 E2 (2) - 2018 ... about the relationship between seminal zinc and fructose concentration in human sperm is scare Therefore, the purpose of this study was to determine the association between fructose and zinc concentration. .. fructose concentration (g/l) and sperm progressive motility (%) Zinc concentration and seminal parameters Table shows the following: • The progressive mobility of the low zinc concentration group... in mean non- progressive motility of males with low zinc concentration compared to males with normal zinc concentration (p= 0.19) Table Seminal zinc concentration and motility of the sperm (Mann