Aldoa And The Cs May Be A Potential Sperm Marker For Smoking

Background: Smoking has been proven to be harmful to human health. And many research showed that smoking was closely related to male fertility. It has been demonstrated that smoking could cause sperm chromosome aneuploidy, chromosome structural abnormalities and DNA fragmentation.Research methods: The 7 weeks age of C57/BL6 J mice were treated with cigarette smoke twice a day, 1 hour for each time. The control group were treated without cigarette smoke. We collected the sperm of these mice and extracted the whole proteins to perform 2-DE. Then MALDI-TOF-MS was used to identify the proteins from the results of 2-DE. We selected 5 proteins identified from MS to verify the results of 2-DE and MS via Western-blotting and Real-time PCR to detect the expression level of these 5 genes in protein and m RNA between the two groups. The functions of the proteins expressed differently in two groups were annotated through Blast2 GO. According to the Gene Ontology functional annotation we classified them into groups. To analyze the pathways which these proteins were involved, KEGG has been used.Results: By the result of 2-DE, we found that over 1000 protein spots showed different expression between two groups. There were 38 protein spots that expressions were changed over 1.5 fold. MALDI-TOF-MS was used to analyze these 38 protein spots and 22 proteins identified. In the 22 proteins identified from sperm, 12 and 10 proteins were down- and up- regulated. We detected these five proteins(ALDOA, ATP5A1, SPATC1, CS, and GPX4) from Western-blotting. The results of 2-DE/MS was proved to be identical. So we thought it was thought to be reliable.Therefore, we also detected these five proteins(ALDOA, ATP5A1, SPATC1, CS, and GPX4) in human sperm samples.The pathway analysis of the differently expressed proteins in sperm was used by KEGG. It was found that these proteins were mainly enriched in energy metabolism.Conclusion: It was shown that daily cigarette smoke exposure could alter the mouse sperm protein expression profile. Many of the affected proteins had functions related to catalytic activity and associated with energy production in certain pathways. Altered expression of such proteins may lower the ATP supply and impair sperm function and motility. We also identified five proteins in human samples, and two proteins(ALDOA and CS) were significantly altered associated with energy production. ALDOA and CS could perform as potential biological markers for identifying sperm from smoking male, and studying such proteins may aid in better understanding the pathogenic mechanisms underlying infertility smokers.