Résumé
L’irradiation de lignées somatiques diverses, de spermatozoïdes et des cellules embryonnaires au laser HeNe de faible énergie (LFE) agirait sur des aspects divers de leur métabolisme cellulaire. Cette action porterait notamment sur l’activité proliferative et respiratoire, ainsi que la synthèse de l’ATP ou l’afflux intracellulaire de Ca2+. Le but de ce travail est d’examiner les effets de l’irradiation au LFE: 1) sur le pouvoir fécondant in vitro des spermatozoïdes de souris en fécondation in vitro (FIV) et 2) sur le pouvoir fusiogénique des spermatozoïdes humains dans l’épreuve du hamster (Sperm-zona free hamster egg penetration assay, SPA). Dans le système de FIV, des ovocytes de souris ont été incubés pendant 24 heures avec des spermatozoïdes épididymaires homologues irradiés ou non. L’irradiation a accru le pourcentage des œufs fécondésin vitro ainsi que le taux du Ca2+ intracellulaire impliquant ainsi l’activation du transport mitochondrial du Ca2+ et la génération de molécules d’H2O2. Dans l’épreuve de fusiogénicité, des ovocytes dépellucidés de hamster ont été incubés pendant trois heures avec des spermatozoïdes irradiés ou non de 43 hommes investigués pour infécondité. Dans 16 cas de faible fusiogénicité sans irradiation, l’irradiation a augmenté le pourcentage de fusion des spermatozoïdes avec les ovocytes de hamster. Dans 27 cas de fusiogénicité normale, l’irradiation n’a eu aucun effet significatif. Ainsi, si cette irradiation est bénéfique chez la souris en FIV, cet effet bénéfique ne se manifeste chez l’homme en SPA que sur les spermatozoïdes déficients.
Abstract
Introduction
The effects of low energy He−Ne 630 laser (LEL) irradiation on various aspects of cell metabolism, including proliferation, respiration, ATP synthesis and Ca2+ uptake, have been recently recognized in somatic cells, animal spermatozoa and embryonic cells.
Objectives
1- To analyze thein vitro effects of LEL irradiation on the mouse IVF model. 2- To assess the effects of experimental LEL irradiation on human spermatozoa in the sperm-zona free hamster egg penetration (SPA) model.
Material & Methods
1- In a mouse IVF model, pooled oocytes were incubated for 24 h. with LEL-irradiated and non-irradiated epididymal mouse spermatozoa. The percentage of fertilized eggs, intracellular Ca2+ and Ca2+ uptake as well as the effects of reactive oxygen scavengers were compared in the two groups. 2- In the enhanced SPA model, with overnight TEST-Yolk preincubation, LEL-irradiated and non-irradiated ejaculated spermatozoa from 43 infertile men were incubated for 3 h with fresh zona-free hamster eggs. The percentage of eggs penetrated (SPA%) by irradiated or control sperm was compared. Acrosome reaction (AR) and Hemizona Assay (HZA) were also performed on the same ejaculates.
Results
1- In mice, brief LEL irradiation enhanced intracellular Ca2+ influx and increased thein vitro fertilization capacity of spermatozoa. Mitochondrial Ca2+ transport mechanisms and H2O2 appeared to be involved in these LEL effects. 2- In the 43 male patients, AR and SPA were correlated, while SPA and HZA were poorly correlated. In the whole group, the mean SPA% of LEL-irradiated and control sperm were not significantly different: 49.8±39.9 and 51.0±34.9, respectively. No significant change was observed in 50% of cases. Following irradiation, SPA% increased in 25% of cases and decreased in 25% of cases. However, classifying all cases according to the SPA cut-off point of 30% as poor and good sperm in this enhanced SPA model, showed that LEL irradiation significantly increased SPA% only in the poor sperm subgroup (n=16) from 5.5.±8.9 to 23.1±25.2 (p<0.0001). This increase was observed in 50% of these patients. In the good sperm category (n=27), no improvement of SPA% was observed. On the contrary, LEL irradiation decreased SPA% from 76.1±24.4 to 67.6±28.
Conclusion
In a mouse IVF model, LEL irradiation of spermatozoa improved intracellular Ca2+influx and egg fertilization. H2O2 seems to participate in the biochemical cascade transforming light signals into a biological response. In man, using the SPA model, LEL sperm irradiation might improve the penetration capacity only in the case of poor quality sperm, by increasing sperm egg penetration in one half of cases, otherwise LEL is ineffective. Further studies are required to confirm these results, but LEL should not be considered for clinical use until its safety has been proven. (Supported by grants from the Chief Scientist Office, Ministry of Health)
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Soutenu par des subventions du Directeur Scientifique du Ministère de la Santé (Israël).
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Soffer, Y., Lubart, R. & Breitbart, H. L’irradiation des spermatozoïdes au Laser HeNe à faible énergie chez la souris et chez l’homme. Androl. 10, 417–426 (2000). https://doi.org/10.1007/BF03034498
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DOI: https://doi.org/10.1007/BF03034498