The Proteomic Changes Analysis In Cigarette Smoking Mice Testis: Pebp1's Potential Role In Cigarette Smoking Governing Sperm Function

Although cigarette smoking is commonly considered to be animportant risk factor for Sperm Infertilization, it is not clear how sperm infertilization is affected.Proteomics is essential to our understanding of cell function, and the identification of specific biomarkers would provide the clues for understandingthe molecular mechanisms for cigarette smoking governing Sperm Infertilization.In this study, we first compared the morphological differences between the cigarette smoking mouse testis and the corresponding normal mouse testis, Finding the number of sperm in the testis is reduced in the the cigarette smoking mouse testis and the Sertoli cell number among seminiferous tubules is aslo reduced.We also described sperm count and sperm motility in the isolated epididymis. Sperm count is significantly reduced in the cigarette smoking mouse epididymis, and the ability of sperm movement is influenced by the cigarette smoking. This indicates that cigarette smoking may affect Spermatogenesis and sperm function, thus affecting sperm quality.To further explore the effects of smoking on the molecular mechanism in testicular sperm function, this study was designed to use comparativeproteomics technology to find the differentially expressedproteins between the cigarette smoking mouse testis and the corresponding normal mouse testis. We have used the highly sensitive two-dimensional gelelectrophoresis (2-DE) coupled with matrix-assisted laserdesorption/ionization time of flight mass spectrometry(MALDI-TOF–MS) for the identification of proteins differentiallyexpressed in the treated testis and thenormalone. With statistical variance of the cigarette smoking versus normal spot volume ratio within the 95th confidence level(Student's t-test; P<0.05), We have detected differences in abundance of 35 proteins ,which consisted of 7 up-regulated and 28 down-regulatedproteins.To analyze the identified proteins,we chose the DAVID server to classify the proteins. The proteins identified are mainly involved in metabolism, signaling transduction, and cellmotility and cytoskeleton organization. Among them, PEBP1, which is down-regulated in treated testis, was significantlyassociated withspermatogenesis. Therefore, PEBP1 was chosen for western blotting and quantitative real-time RT-PCR analysis. Both PEBP1 protein and mRNA levels are decreased in treated testis, which confirmedthe results found by 2-DE. Down-regulation of PEBP1 in cigarette smoking mouse testis led to hyperactivity of MAPK signaling predicted a worse clinical outcome,which may be a useful biomarker to assess the adverse impact ofcigarette smoking to male reproduction, and provide the clues for understandingthe molecular mechanisms for tobacco smoking governing Sperm Infertilization.