Effect of Lamotrigine on motility and cervical mucus penetration, in human sperm cells.
Marina Boxer1, Gili Band2, Igal Madgar2, Ilan Blatt1
1 Epilepsy clinic, Department of Neurology, Sheba medical centre, Tel Hashomer hospital, Israel
2 Male Infertility Unit, Sheba medical centre, Tel Hashomer hospital, Israel
Several antiepileptic drugs (AEDs) may affect sperm motility in vitro. We previously demonstrated the ability of Carbamazepine (CBZ), Phenytoine (PHT), Valproic acid (VPA) and most recently, Topiramate (TPM) to decrease sperm motility and reduse sperm ability to penetrate cervical mucus (a functional test not studied before in relation to AEDs) in concentrations much higher than those in clinical use.
In the presence study we investigated the effect of Lamotrigine (LTG) on these parameters.
Five mg of LTG was dissolved in 1.5% DMSO, and the solution volume was completed with 800μl PBS. Normal semen from healthy human donors (sperm concentration >20*106/ml, >50% progressive motility) was incubated for 2 hours with several dilutions of LTG. Parameters analyzed included sperm motility (% motile sperm), EC50, grade of motility (from 0 to 5, from immotile to good progressive motility, respectively) and the ability of spermatozoa to penetrate bovine cervical mucus after 10 minutes of contact (dept of penetration). Statistical analysis employed t-test comparisons.
After 2 hours of incubation, all the parameters checked was significantly correlated with LTG dose (p<0.05).
In t-test analysis, LTG significantly decreased sperm motility at concentrations
6250 μg/ml, 3125 μg/ml (p<0.001) and 1562.5 μg/ml (p<0.05), without reaching the EC50.
The grade of motility was also significantly decreased above 781.25 μg/ml (p<0.01).
Cervical mucus penetration was significantly inhibited at concentrations above 390.63 μg/ ml.
Effect of TPM on sperm motility and grade of motility (t=0)
Grade of motility (0-5) vs. TPM concentration
Effect of TPM on sperm motility and grade of motility after 2 hours incubation
Grade of motility (0-5) vs. TPM concentration
Effect of TPM on sperm ability to penetrate bovine cervical mucus, after 2 hours incubation
Cervical mucus penetration was significantly inhibited at concentrations ≥1250 mg/L.
Grade of motility in cervical mucus was significantly decreased at concentrations ≥ 2500mg/L.
LTG impairs sperm motility, quantitatively and qualitatively, at concentrations greater than the therapeutic range. At these concentrations it also impairs the ability of human spermatozoa to penetrate into bovine cervical mucus. Similar to our previous results with other AED's (CBZ, PHT, VPA, and TPM), LTG also impaired the physiological function of spermatozoa at higher concentrations relative to those in clinical use.
Effect of Valproate and Carbamazepine on human sperm motility and cervical mucus penetration.
Few studies have investigated the effect of Valproate (VPA) and Carbamazepine (CBZ) on human sperm, and decrease sperm motility was one of the findings. Our arm was to further study the effect of CBZ and VPA on human sperm motility, including sperm ability to penetrate cervical mucus, functional test not hitherto studied in relation to antiepileptic drugs (AEDs).
VPA was dissolved in PBS, CBZ in 2.5 % ethanol +PBS. Normal semen from healthy donors (sperm count >20 million/ml, >50%motile, progressive type of movement) was incubated for 2hr with several dilutions of VPA or CBZ. Parametrs analyzed included sperm motility (%motile cells), EC 50, motility quality grade (1-6), identification of abnormal motility patterns, and ability of spermatozoa to penetrate into bovine cervical mucus after 10 min contact ( number of cells penetrating, depth of penetration). Statistical analysis was performed using the Wilcoxon Signed Ranks Test.
VPA significantly (p<0,05) decreasd sperm motility at concentration >3125mg/ml, with EC50+12500mg/l. Motility quality grade was decreased at concentration >12500mg/l. Cervical mucus penetration was inhibited at concentration >6250mg/l. Shaking, an abnormal motility pattern, was observed at >12500mg/l.
CBZ significantly (p<0,05) decrease sperm motility at concentration >156mg/l, with EC50+312mg/l. Motility quality grade was decrease at concentration >312mg/l. The shaking pattern was observed at >156mg/l.
Both VPA and CBZ impair sperm motility, quantitatively and quantitatively, at concentration well above the therapeutic range.
These concentrations also impair the ability of human spermatozoa to penetrate into bovine cervical mucus, the composition and properties of which are similar to human cervical mucus. This finding implies impairment of the physiologic function of spermatozoa by high concentration of these AEDs.