Динамика морфологических изменений индекса фрагментации ДНК сперматозоидов у пациентов с варикоцеле

Abstract

Abstract Background: To date remains an urgent problem of apoptosis, and the dynamics of change of the index fragmentation of deoxyribonucleic acid (DNA) in spermatozoa in patients with varicocele. Material and methods: The study included two groups: osnovnaya- patients with varicocele (n = 35) and control and healthy donors (n = 32). The average age of patients in both groups was 30 ± 4,5 years. Studying the degree of DNA fragmentation in the sperm was conducted by the method of vital staining of cells with acridine orange (AO). It takes into account the percentage of the number of sperm with green fluorescence of intact sperm chromatin to yellow, orange and red fluorescence fragmented chromatin. 200 spermatozoa were counted in different fields of view of the microscope and calculate the percentage of cells with green and red glow of chromatin. For reversible inhibition of DNA breaks in the sperm were incubated in a refrigerator at 4° C for 24 hours. Accounting chromatin fragmentation was also carried out after staining sperm acridine orange method described above and subsequent microscopy preparation on fluorescence microscope. Results: Patients of the main group affected by varicocele by incubation for 1 hour DNA fragmentation index in the color AO was 19,0 ± 3,0%. Later, after 24 hours of incubation sperm to study at a + 4 оС figure dropped to 10,1 ± 2,0% (p <0.05). In the control group of healthy donors hour incubation of sperm DNA fragmentation index was 2,0 ± 0,5%. After a 24 hour incubation in the cold in this group the rate was 1,5 ± 0,5% (p> 0.05). Conclusions: Thus, this technique sperm incubation at +4 о С can be used to determine sperm count with complete and irreversible double-longitudinal break DNA molecules occurs during the late stages of apoptosis.