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

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

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

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

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

1 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

2 Method

Measuring the concentration of

fructose and zinc

After semen analysis, samples were

centrifuged at 1500 x g for 10 min and

zinc and fructose concentrations assayed

from the supernatant (i.e seminal

plasma) Zinc concentration was assessed

using spectrophotometry (5- Br- PAPS

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

3 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 1 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

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 1 Seminal fructose and some characteristics of the semen

(Independent sample T – test)

Normozoospermia Mean± SD Oligozoospermia Mean± SD fructose test p-value of

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

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 2 and Figure 3)

Figure 1 Correlation between seminal fructose concentration (g/l)

and sperm concentration (billion/ml)

Figure 2 Correlation between seminal fructose concentration (g/l)

and sperm vitality (%)

Figure 3 Correlation between seminal fructose concentration (g/l)

and sperm progressive motility (%) Zinc concentration and seminal parameters

Table 2 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

• 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 2 Seminal zinc concentration and motility of the sperm

(Mann – Whitney test)

Low zinc concentration (n = 84)

Normal zinc concentration (n = 96)

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 4 Correlation between seminal zinc concentration (g/l)

and sperm progressive motility (%) (r = 0.596; 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

Figure 5 Correlation between seminal zinc concentration (g/l)

and sperm immotile (%) (r = - 0.527; p < 0.01) ( Spearman test)

(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

(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 2

forms available for sperm cells (free zinc and

zinc-citrate complex) During ejaculation,

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

- 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

1 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.

2 Biswas S., Ferguson K.M.,

Stedronska J et al (1978) Fructose and

hormone levels in semen: their correlations

with sperm counts and motility Fertility and Sterility, 2, 200 - 204.

3 Aumuller G, Riva A (1992)

Morphology and functions of the human

seminal vesicle Andrologia 24(4): 183 -

196

4 Lu C.J (2007) Standardization and

quality control for determination of fructose

in seminal plasma Journal of Andrology, 28

(2), 207 - 213.

5 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.

6 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.

7 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.

8 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.

9 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

R.(2005) Effects on the quality of

frozen-thawed alpaca (Lama pacos) semen using

two different cryoprotectants and extenders

Asian J Androl, 7 , 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; 1 –

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 (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)

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 9 (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, 2 (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, 6

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.

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

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 double-blind, 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.