Study Of The Relationship Between SLC22A14?SAPG6 And Idiopathic Asthenozoospermia

In recent years,due to factors such as environmental pollution and high living pressure,the incidence of infertility is increasing day by day.Asthenospermia?azoospermia and teratozoospermia are important diseases that affect the reproductive health of men.Among them,One of the most common causes of male infertility is idiopathic asthenozoospermia.It is caused by many factors,including urinary system infections,cryptorchidism,varicocele,and other organic diseases.It also includes late nights,smoking,drinking,and stimulating diets and other bad lifestyles.Asthenospermia is a complex disease caused by a variety of genetic and environmental factors,and its underlying causes remain unclear.It has been determined that some specific gene mutations are related to asthenospermia,but other factors that cause defects in sperm motility in men still need to be explored and studied.Asthenospermia(asthenozoospermia AZS)is characterized by low motility and decreased forward motility.According to the 2010 WHO(World Health Organization,WHO)the new standard of the parameters of semen analysis,sperm density over15�10~6/ml,sperm progressive motility rate is more than 32%for the normal activity of semen,sperm density over 15�10~6/ml,sperm progressive motility rate is less than32%for asthenospermia.Asthenospermia,which is unclear in etiology and pathogenesis,is called idiopathic asthenospermia.The motion of the sperm flagellum is the source of sperm vitality.The principle is that the double microtubules inside the flagellar internal axon are continuously separated and combined under the action of ATP,causing slippage between double microtubules;A variety of accessory structures around the microtubule form a sliding resistance,transforming the longitudinal force generated by the arm into a transverse bending force,causing the flagella to form a wave that oscillates,causing the sperm to move forward,thereby penetrating the cervical mucus to the site of fertilization,and then Pass through the zona pellucida and enter the egg cell to complete the fertilization process.Due to the high complexity of spermatozoa,any change in external factors or internal factors may cause abnormalities in flagellar structure and energy metabolism,affect sperm movement,and lead to male fertility defects.At present,the research on genes and proteins related to male idiopathic asthenospermia is more extensive.In order to further understand the pathogenesis and pathway of idiopathic asthenospermia in men,we used RT-PCR and Western Blot as two experimental methods to detect the expression of SLC22A14 and SPAG6 mRNA and protein in spermatozoa from idiopathic asthenospermia group and normal group.To explore the expression of SLC22A14 and SPAG6 in spermatozoa,thus providing assistance for the diagnosis and treatment of male reproductive health.Materials and methods1 ObjectiveCollected 50 cases of semen in patients(age 22 to 37)with idiopathic asthenozoospermia were treated with idiopathic asthenozoospermia group in the Third Affiliated Hospital of Zhengzhou University from September to November2016,and 50 cases of normal control group in healthy sperm donors(aged 20 to 39)in Human Sperm Bank of Henan Province during the same period.The subjects were satisfied with all kinds of infectious diseases,good physical condition and abstinence2-7d.The semen was collected by masturbation method,then dried in a wide sterile cup,incubated in a 37 degree water bath,and completely liquefied in 30-60min.The sperm concentration in the normal control group was?50�10~6/ml,the percentage of sperm in the forward movement was?32%,the sperm concentration in the idiopathic asthenospermia group was?50�10~6/ml,and the percentage of sperm in the forward movement was<32%.2 Methods2.1 RT-PCRRT-PCR was used to detect the expression of sperm SLC22A14 and SPAG6mRNA in the normal control group and idiopathic asthenospermia group.2.2 Western blottig testsThe expression of SLC22A14 and SPAG6 protein in normal control group and idiopathic asthenospermia group by Western Blot.3 Statistics analysisData were analyzed by SPSS version 21,and the data measured in the experimental results were expressed in x�s.The expression of SLC22A14?SPAG6mRNA and protein in the normal control group and the weak sperm group were compared using two independent sample t tests.Pearson correlation coefficient(r)was used to describe the correlation between the expression of SLC22A14 and SPAG6and the ability of sperm forward movement.The difference was statistically significant in P<0.05.Results1 Expression of SLC22A14mRNA in normal control group and idiopathic asthenospermia groupThe relative expression levels of SLC22A14 mRNA in the two groups of sperm were 0.77�0.08 and 0.53�0.10,respectively.The results showed that the relative expression of SLC22A14 mRNA in idiopathic asthenospermia group was lower than that in the normal control group(t=12.834,P<0.01).2 Expression of SLC22A14 protein in normal control group and idiopathic asthenospermia groupThe relative expression levels of SLC22A14 protein in the two groups of sperm were 0.80�0.09 and 0.55�0.10,respectively.The results showed that the relative expression of SLC22A14 protein in spermatozoa from the idiopathic asthenospermia group was lower than that in the normal control group(t=12.884,P<0.01).3 Expression of SPAG6mRNA in normal control group and idiopathic asthenospermia groupThe relative expression levels of SPAG6 mRNA in the spermatozoa of the two groups were 0.77�0.06 and 0.52�0.10,respectively.The results showed that the relative expression of SPAG6 mRNA in spermatozoa from the idiopathic asthenospermia group was lower than that in the normal control group(t=13.755,P<0.01).4 Expression of SPAG6 protein in normal control group and idiopathic asthenospermia groupThe relative expression levels of SPAG6 protein in the sperms of the two groups were 0.78�0.09 and 0.56�0.09,respectively.The results showed that the relative expression of SPAG6 protein in spermatozoa from the idiopathic asthenospermia group was lower than that in the normal control group(t=12.257,P<0.01).5 The correlation between the expression level of SLC22A14 and SPAG6in sperm and the forward movement of spermSLC22A14 mRNA and protein expression levels were correlated with sperm forward motility(r=0.761,P<0.05;r=0.748,P<0.05),SPAG6 mRNA and protein expression levels also were correlated with sperm forward motility(r=0.806,P<0.05;r=0.766,P<0.05)ConclusionsSLC22A14 and SPAG6 are related to the motility of male sperm and may be involved in the occurrence of idiopathic asthenozoospermia.