SVIBOR - Project code: 3-01-149

MINISTRY OF SCIENCE AND TECHNOLOGY

Strossmayerov trg 4, HR - 10000 ZAGREB
tel.: +385 1 459 44 44, fax: +385 1 459 44 69
E-mail: ured@znanost.hr

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Project code: 3-01-149


BIOCHEMICAL PARAMETERS IN SPERM FUNCTION ASSESSMENT


Main researcher: GAVELLA, MIRJANA (13190)



Assistants
Type of research: basic
Duration from: 01/01/91. to 01/01/95.

Papers on project (total): 13
Papers on project quoted in Current Contents: 6
Institution name: Institut za dijabetes, endokrinologiju i bolesti metabolizma "Vuk Vrhovac", Zagreb (45)
Department/Institute: department of laboratory medicine
Address: Dugi Dol 4a
City: 10000 - Zagreb, Croatia
Communication
Phone: 385 (0)1 2332-222
Fax: 385 (0)1 2331-515

Summary: Original study was carried out to define at biochemical level the nature of the lesions causing sperm function impairment and subsequently the impairment of sperm fertility potential.The etiology of defective sperm function was studied and results obtained as follows: 1) - alteration of specific activity of NADH-dependent oxidoreductase(diaphorase),an enzyme located in the middle part of the spermatozoa and involved in sperm metabolism. The manyfold increase in the activity of this enzyme in oligozoospermic patients indicates higher proportion of defective sperms which retained cytoplasmic material during spermiogenesis. 2) -relationship between diaphorase, isozyme LDH-C4 and superoxide anion. A positive correlation between superoxide anion production and diaphorase activity indicates that only sperms with negligible superoxide anion generation and low diaphorase activity can be considered as functionally competent.3) - relationship between reactive oxygen species generation and the functional competence of human spermatozoa.The observed in vitro effect of superoxide anion on sperm membrane measured by ability of hypoosmotic sperm tail swelling points to the changes of functional integrity and membrane fluidity of sperms resulting in decreased fertility potential.4) - inhibitory effect of some drugs on the excessive superoxide anion generation.Superoxide anion production was reduced by 29-72% following the addition of 10 mM pentoxifylline which did not depend on the initial superoxide production by sperm.The significant inhibition of superoxide anion release with lower concentration of 3.7 mM pentoxifylline in sperm prepared for in vitro fertilization procedures was also established. These results point to a potential use for pentoxifylline a) in the treatment of male infertility caused by excessive reactive oxygen species production and b)in improvement of sperm preparation for in vitro fertilization. 5) -stimulatory effect of pentoxifylline or experimentally induced lipid peroxidation of sperm plasma membrane. Induction of lipid peroxidation in the presence of pentoxifylline and ferrous/ascorbate ions promotors may have an adverse effect on the quality of sperm suspensions prepared for in vitro fertilization.

Keywords: human sperm function, fertilizing potential, oxidative stress, reactive oxygen species, superoxide anion, lipid peroxidation, pentoxifylline, diaphorase, lactat dehydrogenase isoenzyme LDH-C4.

Research goals: The study was aimed to obtain relevant knowledge on the mechanism determining male fertility and/or infertility. From the viewpoint of an infertile couple and their andrologist,there is no firm background for diagnosing and introduction of adequate therapy.Despite attempts to solve this problem pragmatically, it is evidenced today that the determination of male infertility depends not only on possible sperm number deficiency but also on the deficiency of the functionally competent sperms.Additionally,there is not enough knowledge on biochemical and biological functions of semen components and their relationship.It is quite positive,however, that these are the important factors by which fertility,i.e. infertility can be affected. Therefore this study was carried out to identify those sperm function deficiences which could serve as specific biochemical markers of sperm quality. The study was aimed to evidence a) alterations of some enzymes participating in the metabolic pathways to provide energy necessary for sperm motility b) sperm membrane alterations as the results of sperm exposure to oxidative stress and c) new prospects in developing adequate therapy arise by defining some biochemical alterations in case of functional sperm defects.Therefore,in vitro effects of potential agents which could affect the superoxide anion generation, such as pentoxifylline,were investigated.


COOPERATION - INSTITUTIONS


  1. Name of institution: Institut "Rudjer Bošković" Odjel za eksperimentalnu biologiju i medicinu
    Type of institution: University/Faculty
    Type of cooperation: Joint publishing of scientific papers
    City: 10000 - Zagreb, Croatia

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Last update: 09/27/95
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