Expression Of COX6B1 In Weak Spermatozoa And Its Preliminary Study On In Vitro Fertilization Outcome

Objective The aim of this study was to investigate that in mature sperm,cytochrome c oxidase 6b1 m RNA levels and protein expression differ between asthenospermia patients and normozoospermia male,and comprehend the protein positioning in the sperm. Preliminary discussion the relevance between COX6B1 and asthenospermia in sperm.Methods Collected 50 semen samples from asthenospermia patients group and normozoospermia male group,Pure Cepition density gradient centrifugation was used to separate the sperm,extracted Total RNA of sperm andreverse transcripted to c DNA.Designed COX6B1 gene-specific amplification primers,and through the method of Real-time fluorescence quantitative reverse transcription polymerase chain reaction(q RT-PCR) and compared the COX6B1 m RNA relative expression levels.Diluted sperm into proper concentration and made sperm smears.We used immunofluorescence technique and interpretated the position of COX6B1 in mature spermatozoa under 600×oil immersion of laser scanning confocal fluorescence microscopy.Selected random view and took photos,then analyzed the difference of average fluorescence intensity between the two groups.Results The COX6B1 located in the middle of the sperm tail.The asthenospermia patients group m RNA expression were down regulated and the m RNA relative expression was significantly lower than the normozoospermia male group(1.393±1.447 vs 4.249±4.933,P=0.008<0.01).And the average fluorescence intensity COX6B1 protein in asthenospermia patients group was lower than the control group(32.341±6.195 vs 48.931±5.784),the difference was statistically significant.Conclusion The COX6B1 m RNA levels and protein expression in asthenospermia patients group were lower than the normozoospermia male group.It suggests that the COX6B1 gene is expected to be a molecular marker of the asthenospermia.Objective Compared the the outcome of IVF in asthenospermia patients group and normozoospermia male group with COX6B1 differential expression.Methods The whole 100 couples in two groups accepted in vitro fertilization and embryo transfer in the Reproductive Center of the First Affiliated Hospital of Guangxi Medical University.Fertilization and clinical pregnancy outcome in the two groups were compared.Results The male age, the female age and the years of infertility there were no significant difference in two groups.The fertilization rate in asthenospermia patients was significantly lower than the normozoospermia male group(72.36% vs 79.91%,p<0.01) and the normal fertilization rate showed the same trend.The cleavage rate, excellent embryo rate, clinical pregnancy rate and early abortion rate there were no statistically significant difference in two groups(P>0.05).Conclusion The fertilization rate and the normal fertilization rate in asthenospermia patients was significantly lower than the control group and there were no statistically significant difference about the cleavage rate, excellent embryo rate, clinical pregnancy rate in two groups.It was speculated that the decrease of COX6B1 protein may have a negative effect on the energy metabolism of mitochondria, thus reducing the fertilization rate.