активности Nа+,+-АТРазы: в первом случае Ki составляет 20–30 нM, а во втором (для уабаина) — 100–120 мкM.
Установлено, что концентрация нитритов в суспензии сперматозоидов не меняется в условиях олигозооспермии, относительно нормоспермии, но увеличивается в 2 раза при астенозооспермии (снижении подвижности сперматозоидов).
В ходе выполнения работы разработан новый чувствительный биохимический тест на наличие патологии олигозооспермии. В основе метода лежит определение активности “общей” эозинчувствительной АТРазной активности сперматозоидов, которая при добавлении 0,5раствора сапонина, в норме увеличивается в 3,8–4 раза (изменения относительно контрольного значения без детергента достоверны), а в условиях развития олигозооспермии — только в 1,3–1,5 раза (изменения не достоверны).
Ключевые слова: активность Са2+,2+-, Nа+,+-, Н+- и “базальной” Mg2+-АТР-гидролаз, каликсарены, нитрит-ионы, сперматозоиды, эякулят, олигозооспермия.
KocheshkovaIdentification and properties of ion-transporting ATP-hydrolases of man’s spermatozoa in the conditions of oligozoospermia. —Manuscript.
PhD thesis for obtaining the scientific degree of PhD (Biology) in the speciality 03.00.04 — biochemistry. Institute of Animals Biology, Lviv, 2007.
Dissertation is devoted to the identification of ATP-hydrolasing systems, which have extra- and intracellular localizations of their active centres in man’s spermatozoa. It was studied their properties, peculiarities of functioning in normal and oligozoospermia conditions, and also established the changes of concentration of nitrite-ions under development of this pathology.
It was shown that oligozoospermia is a widespread pathology of the male reproductive system (18all of the samples). There was defined the optimal regime of reaction of Mg2+-dependent enzymatic hydrolysis of АТР in suspension of spermatozoa.
It was identified several ‘extra’- and ‘intro’-АТР-hydrolysing systems in male gamete’s membranes. The values of Са2+-ATPase activity of endoplasmic reticulum increased significantly in the case of oligozoospermia. Contrary to this, the activity of Са2+-independent ATPase systems (Nа+,+-, ‘basal’ Mg2+- and Н+-АТРases) decreases in case of oligozoospermia. We calculate main kinetic indexes (constant of inhibition Ki and Hill’s coefficient nH) for selective (tapsigargin, ouabain, sodium azid) and non-selective (eosininhibitors of proper ATPases systems of men’s spermatozoa by linearization in the Hill’s co-ordinates of curves of catalytic titration.
The calixarenes C97 and C107 inhibited the enzymatic activity of Na+,K+ATPase more effectively than ouabain. Content of nitrite-ions in spermatozoa doesn’t change in case of oligozoospermia, but correlates positively with development of asthenozoospermia.
There was suggested new biochemical diagnostic test for oligozoospermia. It is based on the increase of eosinATPase activity in 3,8–4 times in health men’s spermatozoa (comparing to the control without detergents) and the absence of its significant increase in the case of oligozoospermia.
Key words: activity of Са2+,2+-, Nа+,+-, Н+- and ‘basal’ Mg2+-ATP-hydrolases, calixarenes, nitrite-ions, spermatozoa, ejaculate, oligozoospermia.