| Background and object: The incidence of male infertility is increasing year by year,and the factors causing male infertility varied,including varicocele is one of the most common causes.Although the clinical doctors recognized that varicocele will affect the fertility of patients significantly,but its specific mechanism of causing infertility is still not clear.At present,it has been found that varicocele is an obstacle to testicular temperature regulation,leading to the chronic hypoxia and excessive production of the reaction oxygen.Proteomics is an advanced research method,and it can be a good study of cell or tissue in protein function and metabolic process at the molecular level.In the different disease states,isobaric tags for relative and absolute quantification(i TRAQ)has been widely used in the research of the male infertility proteins with a highly value.Seminal plasma is composed of lipids,carbohydrates,a large number of peptides and proteins.And seminal plasma is secreted by male prostate,seminal vesicle,bulbourethral gland,epididymis and other glands.High concentration of the tissue-specific proteins in the seminal plasma can be used as the biomarkers of the male fertility assessment.This study is based on the previous experimental study,in order to analyze the seminal plasma protein proteomics.The study will find out the protein which has significant differences.Analyze the molecular function of the differential proteins and explore the mechanism of infertility induced by varicocele.Then identify the differential proteins by western blot.Finally,the study can obtain the biomarkers of varicocele-induce infertility,and can provide support for clinical diagnosis and treatment.Methods: Collect 20 samples of semen from the left varicocele infertility patients and 20 samples of normal male semen.All the seminal plasma of the collected semen samples were extracted as the experimental samples.The experimental samples were labeled with the i TRAQ kit,117 labeled the varicocele infertility group,and 114 labeled the normal group.Then analyzed the samples by proteomics.The data was analyzed by Proteome Discoverer 1.3 software.The protein which up multiple ? 1.5,down multiple ? 0.7,P ? 0.05 was considered significantly statistical difference.The differential proteins were enrich-analyzed by the molecular of gene ontology,and can be used as the infertility marker.Testing both varicocele infertility group and normal group by western blotting to verify the differentially expressed proteins.Use the image pro plus 7.0 software to analyze the grayscale of the resulting image.Results: Totally,39 differentially expressed proteins were identified,including 22 up regulated proteins and 17 down regulated proteins.Molecular function analysis showed that there were 14 kinds of molecular functions of differential proteins,which were mainly involved in protein binding,catalytic activity and structural activity.C2CD2 up regulated 2.532 times,EML1 down regulated 2.162 times,LIM3 down regulated 0.544 times.Western blot verified that C2CD2 and LIM3 were significantly higher than those in normal group and EML1 was lower than that in normal group.Conclusion: Varicocele mainly affects male infertility in molecular function through protein binding,catalytic activity and structural molecular activity.There is a significant correlation between varicocele and the abnormal expression of C2CD2,LIM3 and EML1.Also C2CD2,LIM3 and EML1 can be regarded as the infertility markers of varicocele patients.ITRAQ in an effective tool for screening infertility-related proteins in varicocele patients. |
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