In this RCT study, the demographic variables
of patients in two
groups of three and two- drug treatment were similar in terms of age (27± 2 vs.
28± 2), duration of infertility (4.5± 0.6 vs. 4 ± .5), and BMI (22.6 ± 1.7 vs.
23.4 ± 2) respectively.
results of comparison variables in control and treatment groups are listed in
table 1, regarding global DNA methylation, DNMTs expression, Sperm
chromatin and DNA integrity tests results and sperm parameters. Sperm
concentration in both three and two-drug treatment groups showed a significant
increase compared with pretreatment (p= 0.001 vs. p= 0.035) respectively, while
it was still significantly lower compared to control group. After the
three-drug treatment, progressive motility and normal morphology improved
significantly compared to pretreatment, such improvement did not occur in the two-drug
group. Sperm viability showed a significant increase after the three-drug
treatment period consistently comparable with the control group (p=0.02 vs.
p=0.07), while this improvement as a result of treatment was not observed in
the two-drug treatment group.
Based on the results of the Aniline blue+ results, sperm chromatin excess
of histone showed a significant reduction after the three-drug treatment
compared with the pretreatment and unlike the two-drug treatment group.
However, the level of dark blue stained spermatozoa in the three-drug treatment
group was still significantly higher than in the control group. A high level of
chromatin condensation was obtained after both three-drug and two-drug treatment
consequently by the Toluidine blue+ results (p=0.001). The rate of fragmented
DNA sperms with TUNEL+ test decreased significantly in both
treatment groups (p=0.001), but it was still significantly higher than the
expression level of DNMT3A and DNMT3B genes after treatment in
two and three-drug group showed no significant difference compared with
pretreatment. Global methylation decreased significantly after the three-drug
treatment (p=0.02), while there was still a significant difference in methylation
level in the control group. However, sperm global methylation showed no
significant difference in the two-drug treatment.
sperm concentration was inversely correlated with DNMT3B transcription. No
significant correlation was observed between progressive sperm motility, normal
morphology and sperm viability with expression levels of the DNMTs genes.
The TUNEL+ results were directly related to the DNMT3A and DNMT3B
transcription. Global methylation level was negatively associated with the Sperm
concentration, progressive motility and normal morphology (p?0.001), but global
methylation was positively related to the results of TUNEL+,
Aniline blue+ and Toluidine blue+ results (p?0.001).