The Correlation Between Semen Elastodextrase,sperm DNA Fragment And Main Parameters Of Routine Semen Analysis

Background:In recent years,along with changes of society,environment,living habits and other factors,the male infertility rate shows an obvious upward trend.According to the world health organization(WHO),about 15% of couples worldwide suffer from infertility,of which about 20% percent are caused solely by male factors,and about 25% are caused by both male and female factors.Therefore,improving the examination for male infertility has become an urgent need in clinical practice.Reproductive tract infection can affect the quality of sperm,but it is difficult to find a referenced threshold to judge the impact of inflammation on fertility capacity.However,elastase of spermatoplasm is distributed in a variety of cells and tissues,such as neutrophils and macrophages,and can play a local anti-inflammatory effect together with other oxidation substances,and it is evenly distributed in semen.ELISA can be used for batch detection,with little difference,high reproducibility and high accuracy,and thus suitable for clinical use.In addition,some studies have pointed out that,latent infection and dominant infection can be distinguished according to the difference of elastase value,which can provide more accurate evidence for deciding the correct treatment of patients.During the treatment of assisted reproduction,a considerable number of patients failed to find abnormalities in routine or biochemical semen examination.Therefore,researchers are paying more and more attention to explore the internal causes of sperm.DNA is the carrier of genetic information and plays a decisive role in human reproduction and expression of inherited traits in offspring.Sperm DNA damage can reduce sperm fertilization and embryo development potential,thereby affecting the pregnancy rate of natural pregnancy or assisted reproduction and may increase the abortion rate.Therefore,it is of great clinical significance to analyze the relationship between sperm DNA damage and semen routine parameters and spermatozoa biochemistry,to explore the influencing factors of sperm DNA damage and its application value in male infertility diagnosis.Objective:To explore the correlation between semen elastodextrase,sperm DNA fragment and main parameters,acrosomase and anti-sperm antibody in semen routine examination,so as to provide more accurate basis for diagnosis and treatment of male infertility.Method:(1)A total of 464 semen samples of male patients admitted to the Reproductive Center of the Third Affiliated Hospital of Guangzhou Medical University from July 2017 to April 2018 were collected,and relevant clinical data were sorted out.(2)Routine semen tests were conducted in accordance with the standard operating procedures of the WHO Laboratory Manual for Human Semen Examination and Processing,5th Edition.(3)The percentage of anti-sperm antibodies in spermatozoa was detected by the mixed anti-globulin reaction.(4)The concentration of elastomeric dextrase in seminal plasma was detected by enzyme linked immunosorbent assay.(5)The sperm DNA fragment index was detected by flow cytometry-acridine orange method.(6)The Sperm acrosomase activity was detected by modified Kennedy method.(7)According to the semen routine results,464 samples were divided into normal and abnormal groups,and other parameters between the two groups were analyzed for differences and correlations.The samples were also divided into normal and abnormal groups using the sperm DNA fragment index of 30% as the cut-off level,and the differences and correlations of other parameters were analyzed between the two groups.According to the concentration of elastase in seminal plasma,the patients were divided into negative group(n=262): group A(< 290 ng/ml),inapparent infection group(n=120): group B(290-1000 ng/ml),confirmed infection group(n=82): group C(> 1000ng/ml).Results:(1)Comparing the normal and abnormal semen group,there were significant differences in age,ALH,DNA fragment index(DFI)and acrosomase content between the two groups(P<0.05),among which age(r =-0.092,P<0.05)and DFI(r =-0.268,P<0.05)were negatively correlated,ALH(r = 0.104,P<0.05)was positively correlated with acrosomase content(r = 0.227,P<0.05).Other parameters(VAP,VCL,VSL,LIN,STR,BCF,elastase,anti-sperm antibody(AsAb),leukocyte)were not correlated between the two groups(P>0.05).(2)Comparing Seminal plasma elastase negative group(group A)and the recessive infection group(group B)and confirmed infection group(group C),there are statistically significants among the three groups of age,PH,acrosin,AsAb and leukocyte(P < 0.05),There are no significant differences among the three groups(P>0.05)of the other parameters(volume,concentration,PR,PR+NP,survival rate,sperm abnormality rate,VAP,VCL,VSL,LIN,STR,ALH,BCF,DFI).The parameters of the statistical differences between the three groups of the two further comparison,there are statistically significant between the negative group and the recessive infection group of PH,acrosin between the negative group and the recessive infection group of acrosin,between negative and confirmed infection group of AsAb,between the negative group and infection group of leukocyte.Age(r = 0.127,P<0.05)and PH(r = 0.206,P<0.05)were positively correlated with the concentration of spermatic elastase,while acrosin(r =-0.105,P<0.05)was negatively correlated with the concentration of spermatic elastase.(3)Comparing the abnormal and abnormal groups,age,sperm concentration,PR,PR+NP,survival rate,abnormality rate,VAP,VCL,VSL,ALH,acrosin and AsAb showed statistically significant differences between the two groups(P<0.05).Age(r = 0.211,P<0.05),abnormality rate(r = 0.282,P<0.05),VAP(r = 0.106,P<0.05),VCL(r = 0.101,P<0.05)and VSL(r = 0.116,P<0.05)were positively correlated with DFI,while PR(r =-0.59,P<0.05),NP+PR(r =-0.585,P<0.05),sperm survival rate(r =-0.529,P<0.05),and acrosin(r =-0.177,P<0.05)were positively correlated with DFI.The other parameters(volume,PH,LIN,STR,ALH,BCF,spermatic elastase,leukocyte)were not correlated between the two groups(P>0.05).Conclusion:1.The level of elastase in spermatic plasma is positively correlated with age.Increasing the level of elastase can increase the PH value of semen,but has no effect on other results of semen routine.The increased level of elastase in spermatoplasm can lead to decreased activity of sperm acrosomase and increased level of anti-sperm antibody.2.Sperm DNA damage is positively correlated with age and sperm malformation rate.Increased sperm DNA damage can reduce sperm concentration,activity,and survival rate.In addition,increased sperm DNA damage can also lead to decreased sperm acrosomase activity and increased anti-sperm antibody levels.3.Male reproductive age should be paid attention to.Age has an important impact on sperm function,and is a risk factor for the increase of spermatic elastase level and sperm DNA damage.4.Spermatic elastase and sperm DNA damage have significant negative effects on semen quality,which is of great significance for evaluating male fertility,and can be used as a supplement to routine semen examination.Spermatic elastase can be used as an indicator of reproductive tract infection,and sperm DNA damage significantly reduces sperm motility.It is suggested that patients with asthenospermia should pay special attention to sperm DNA damage detection,and make corresponding treatment plans according to DFI value.