Antioxidant’s Protection Of Sperms From Oxidative Stress Injury In Centrifugal Process

Objective: To investigate determination of semen reactive oxygen speciesconcentration and semen parameters and their relationship with the outcomes ofin vitro fertilization.Analysis whether there is a correlation between semenreactive oxygen species and semen quality,the outcomes of in vitrofertilization.Assess the value of the semen reactive oxygen species in thediagnosis and treatment in male infertility.Methods: Data from October2012to January2013of72couples due totubal factor infertility who accepted the first IVF treatment in our center.Rigorous analysis of sperm morphology by using Diff-Quik sperm morphologyanalysis, using aniline blue staining for detecion of sperm nuclear maturity.The semen reactive oxygen species contents of72cases infertile men weredetected by Chemiluminescence method．Observe the fertilization on the firstday after in vitro fertilization and cleavage on the third day.Pearson correlationanalysis was used to analyze the relationship between semen reactive oxygen species content and sperm quality,outcomes of in vitro fertilization.Results: The semen reactive oxygen species contents of72cases infertilemen were (72.19±3.56)RLU/s. Pearson correlation analysis showed that thesemen reactive oxygen species content was negatively correlated with spermmotility rate, normal morphology rate, IVF fertilization rate and normalfertilization rate (r=-0.799,-0.827,-0.728and-0.792; P=0.012、0.032、0.000、0.003，all P<0.05)；and it was positively correlated with sperm density and theimmature sperm rate (r=0.317,0.794; P=0.008、0.001，all P<0.05)．However，no correlation was found between the semen reactive oxygen species contentand the age,semen volume,cleavage rate,good quality embryo rate(all P>0．05).Conclusions: There is a correlation between semen reactive oxygen speciesand semen quality, the outcomes of in vitro fertilization. The right amount ofsemen reactive oxygen species content can promote the occurrence, maturationand the capacitation mature of sperm．The excessive amount of reactive oxygenspecies can decrease sperm motility, fertilization rate；but it has no significantimpact on the IVF cleavage rate and good quality embryo rate． Objective: To study the influence of normal sperm and oligoasthenozoosp-ermia (OAS) during in-vitro process of centrifugation. To explore the protectionrole of L-carnitine and Glutathione for sperms oxidative stress injury during in-vitro process of centrifugation.Methods: According to the WHO standard,35normal sperm samples and38OAS samples were selected and1800ul was taken from each sample.Thesamples were divided into control group, centrifugal control group, centrifugaland GSH group, centrifugal and L-carnitine group, with450ul in each group.In the control group and centrifugal control group,only EBSS balanced solutionwere added.In the centrifugal and GSH group, EBSS was added with a certainconcentration of GSH.In the centrifugal and L-carnitine group, EBSS wasadded with a certain concentration of L-carnitine. Each treated sperm wasdetected with reactive oxygen species(ROS), malondialdehyde(MDA)concentration and DNA fracture index(DFI). Then we did the analysis ofvariance of randomized block design and multiple comparison.Results:1.The results in the four groups showed significant statiststical differences. The normal sperm: FROS=280.10,P=0.000；FMDA=15.98,P=0.001；FDFI=78.84,P=0.002;all P<0.05).The oligoasthenozoospermia sperm: FROS=9.45、P=0.000;FMDA=15.79,P=0.000;FDFI=13.56,P=0.000,P<0.05).2.Both in oligoasth-enozoospermia sperm and normal sperm,ROS,MDA and DFI levels ofcentrifugal control group were significantly increased than control group(allP<0.05). The ROS, MDA and DFI levels in the GSH group and L-carnitinegroup were significantly lower than the control group (all P<0.05).However,theresults showed no significant difference between the GSH group and L-carnitinegroup (P>0.05).Conclusion: All the oligoasthenozoospermia sperm and normal spermwill be Produced excessive reactive oxygen species in the centrifugalprocess,which will be resulted in peroxidative damage.Adding a certainamount concentration of GSH or L-carnitine before semen centrifugal canreduce the production of the excessive reactive oxygen species, peroxidativedamage and improve the sperm quality.