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Human sperm quality and lipid content after migration into normal ovulatory human cervical mucus containing low numbers of leukocytes

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Asian Journal of Andrology 2009年第3期11卷 308-316页

作者： nozha chakroun-feki Patrice Therond Martine Couturier Florence Eustache Gerard Limea Alain Legrand Pierre Jouannet Jacques Auger Laboratory of Reproductive Biology/CECOS （Center of Study and Conservation of human Eggs and Sperm） Cochin Hospital Rend Descartes University Paris 75014 France Laboratory of Clinical and Metabolic Biochemistry EA 3617 Rend Descartes University Paris 75270 Cedex 06 France INSERM U551 La Pitie Hospital 75651 Paris Cedex 13 France

The aim of this study was to investigate whether a relationship exists between the presence of low numbers of leukocytes in normal ovulatory cervical mucus and sperm quality and lipid content after migration. The percentages of live, motile and morphologically normal spermatozoa, movement parameters assessed by computer-aided sperm analysis （CASA）, and ionophore-induced acrosome reaction measured by flow cytometry were determined before and after migration. High-performance liquid chromatography with ultraviolet detection was used to measure the sperm lipid content, including the various diacyl subspecies. The number of leukocytes found in solubilized mucus samples was counted using a haemocytometric method. Overall, the presence of leukocytes in the cervical mucus samples did not significantly influence sperm motility and morphology, sperm kinematic parameters, or the sperm content in sphingomyelin or cholesterol. In contrast, after migration, the decrease in various sperm diacyls and the level of induced acrosome reaction was significantly less pronounced in mucus samples containing ≥10^4 leukocytes than in mucus samples with no or rare leukocytes whereas the level of induced acrosome reaction was higher. The present data suggest that the low level of leukocytes found in normal ovulatory cervical mucus could influence the process of sperm lipid remodelling/capacitation.

关键词： cervical mucus leukocyte lipid spermatozoa

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Sperm quality improvement after date seed oil in Vitro supplementation in spontaneous and induced oxidative stress

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Asian Journal of Andrology 2009年第3期11卷 393-398页

作者： Ben A. Fatma chakroun F. nozha Dammak Ines Attia Hamadi Hentati Basma Ammar K. Leila Human Pathologies and Oxidative Stress Unit Superior Institute of Biotechnology of Sfax Sfax 3028 Tunisia Laboratory of Histology Embryology and Reproductive Biology Medicine Faculty of Sfax Sfax 3028 Tunisia

In vitro supplementation with date seed oil （DSO） can protect spermatozoa against hydrogen peroxide （HiO2）- mediated damage and can improve sperm function, possibly owing to antioxidant properties. We tested the antioxidant effects of DSO on human sperm motility, sperm viability, reacted acrosome and lipid peroxidation assessed in vitro after H202-mediated oxidative damage in spermatozoa. Sixteen patients （mean age： 35 years; range： 25-45 years） referred to the Histology-Embryology Laboratory of the Medicine Faculty of Sfax for semen analysis after 12-24 months of sexual intercourse without conception were selected. After spermiogram, sperm selection by twointerface discontinuous Sill Select gradient was performed, and selected spermatozoa were used in four experimental assays： control; incubation with 100um H2O2; incubation with 0.1% DSO; and co-incubation with 0.1% DSO and 100 um H2O2. Motility and viability were determined using World Health Organization criteria. Acrosome reaction and lipid peroxidation were assessed by staining with fluorescein isothiocyanate-Pisum sativum and spectrophotometric measurement of malondialdehyde, respectively. Results showed that incubation with H2O2 alone led to a significant increase in lipid peroxidation （57.83%, P 〈 0.05） associated with a significant decrease in sperm motility, sperm viability （after 30 min and 24 h） and percentage of reacted acrosome （P 〈 0.05）. Date seed oil im- proved sperm motility after 24 h of incubation （P 〈 0.05） and protected spermatozoa against the deleterious effects of H2O2 on motility, viability, acrosome reaction and lipid peroxidation. We conclude that supplementation with DSO may have a function in antioxidant protection against male infertility.

关键词： acrosome reaction date seed oil in vitro lipid peroxidation sperm motility sperm viability

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Evaluation of the impact of leucocytospermia on semen oxidative status by chemiluminescence technique in infertile men

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Advances in Bioscience and Biotechnology 2013年第11期4卷 30-40页

作者： Afifa Sellami nozha chakroun Yemna Rtaib Hajer Hdhili Riadh Ben Mansour Louati Dolira Kais Chaabene Leila Keskes Saloua Lassoued Tarek Rebai Gynaecology and Obstetrics Department Hedi Chaker Academic Hospital Sfax Tunisia Laboratory of Cell Culture Biotechnology Institute Sfax Tunisia Laboratory of Histology and Biology of Reproduction Medical School Sfax Tunisia Laboratory of Histology and Biology of Reproduction Medical School Sfax Tunisia Histology Embryology Research Unit Medical School Sfax Tunisia Laboratory of Histology and Biology of Reproduction Medical School Sfax Tunisia Histology Embryology Research Unit Medical School Sfax Tunisia

The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage. The aim of our study was to evaluate the impact of leucocytospermia on basal and FMLP (Formyl-Methionyl-Leucyl-Phenylalanine) induced oxidative status in semen of infertile men. We also analyzed the correlations of the spermatic parameters with amounts of ROS in semen. Our study included 50 semen samples of infertile men. Sperm analysis was performed using WHO standardized method. Seminal leucocytes were quantified using peroxidase technique. The measurement of ROS levels in semen was made by chemiluminescence assay. We measure respectively ROS amounts in neat semen and in washed sperm cells suspension from the same ejaculate. We also applied the test of provocation of leucocytes by FMLP on neat and washed samples to assess the spermatic oxidative status after leucocyte stimulation. Our results showed significant correlations between ROS levels in neat semen and many sperm parameters: motility, sperm concentration, leucocytes concentration and the rate of sperm cytoplasmic droplets. The studied samples were divided into 2 groups: (G1) composed of 36 samples without leucocytospermia and (G2) composed of 14 leucospermic samples. ROS levels were significantly lower in G1 than in G2 (p = 0.002). ROS production was significantly increased after application of FMLP in washed leucospermic samples (p = 0.001). The measurement of ROS in neat semen is a considerable contribution to explore the impairment of semen quality in infertile men. ROS levels in washed semen reflect the oxidative status generated by sperm preparation techniques used in assisted reproductive procedures. Levels of ROS are highly influenced by the presence of leucocytes and associated with decreased seminal param

关键词： Semen Oxidative Stress Leucocytospermia Chemiluminescence ROS FMLP

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