Study The Role And Mechanism Of Differential Expression Sperm Proteins In Asthenzoosperm

Objective1)iTRAQ (isobaric tags for relative and absolute quantitation, iTRAQ)labeling coupled with2D-LC-MS/MS (two-dimensional-liquid chromatography coupled by tandem mass spectrometry,2D-LC-MS/MS) method was used for determining the proteome in the asthenozoosperm patients from the disease degrees of the mild, the moderate and the severe to gain more knowledge about sperm proteome.2) To explore the role of differential expression of proteome in patients of asthenozoosperm by study the change expression of the sperm proteins in the mild, the moderate,the severe asthenozoosperm patients and normal.3) Study the change of sperm proteins of reproduction in proteome to explore it’s change rule of these proteins in asthenozoosperm.Contents and MethodsiTRAQ labeling coupled with2D-LC-MS/MS method was used to study the role of the expressional change of sperm proteome in the asthenozoospermia. First,we collected sperm samples of males include normal and asthenozoospermia patients(the mild, the moderate and the severe); then iTRAQ labeling was used to label sperm proteins and2D-LC-MS/MS was used to study these mix proteins for determining the expressional change of sperm proteome in normal and asthenozoospermia groups. Then, the results were analyzed through some protein website and analysis tools to gain the molecular function and biological process about the differential expression proteins. The role and mechanism of differential expression proteins in asthenozoospermia was analyzed and studied.Results1) Our study collected142sperm samples of males include91normal cases and22mild,18moderate,11severe asthenozoospermia cases. There were no significant differences in age, abstinence times and sperm form of four groups.2)1073proteins were identified and511proteins of them were2times and more expressed in the mild, the moderate and the severe groups than in normal group. These proteins include molecular function related proteins, biological process associated proteins, cellular components, protein classes and pathway related proteins. The most of molecular function related proteins were catalytic activity proteins, binding proteins and structural molecule activity proteins; most of biological process associated proteins include metabolic process, cellular process, immune system process and cell communication; the most of cellular components was intracellular and plasma membrane was less part.3) The significant increase/decrease expression proteins among the mild, the moderate and the severe group were109/193,31/282and38/211respectively; and an obviously different increase/decrease expression model was observed in the three groups(P<0.05). The together decrease/increase expression proteins amounted to104/10in three asthenozoospermia groups.4) There were48proteins associated reproduction process in1073proteins. These reproduction proteins were expressed different among three asthenozoospermia groups.Conclusions1) iTRAQ labeling can be well applied in the asthenozoospermia sperm proteins research, we can get more comprehensive proteome differential expression of information comparably with2-DE.2) In the asthenozoospermia,there are a large numble of sperm protein expression is significantly altered, involving a variety of molecular functions and biological process. Differential expression of sperm protein in molecular function are mainly embodied in catalytic activity, binding and structural molecule activity; in terms of biological process are mainly embodied in metabolic process, cellular process, immune system process and cell communication. Its main expression changes are decrease expression, sperm vitality decline may be associated with reduced expression of closely related proteins.3) Involving the reproductive process of protein in different degree of asthenozoospermia group show different change patterns.