The Screening And Ldentification Of Differential Expression Proteins In Placenta Tissue Of Dairy Cow Retained Fetal Membranes

Retained Fetal Membranes (RFM) is a reproductive disorder disease. It harmed the healthy of dariycow. The cause and influencing factors of this disease is complex. But the mechanism of the diseasewas not definite, especially the molecular basis was unknow, and the control efficiency was low.The comparative proteomics and bioinformatics are good methods in the study of the differentialexpression of proteins in placenta of the cow with RFM. Main pathopoiesis molecules could befound. The differential expression proteins will be screened, evaluationed and verificated by overtwo techniques. The regulating molecule network of protein will be analysised. The finding of keyproteins is important. It can reveal the pathopoiesis and channel, clarity the pathogenesis, screen theprophase clinical diagnosis marker. The molecular therapy of RFM would be possible.Ten placenta tissues were collected from3cows with RFM and3cows with health control. Proteinsin placenta tissue of RFM and healthy underwent two-dimensional polyacrylamidegelelectrophoresis,2D-PAGE and silwer stained gel imaging toget the differential expression proteins,then, the spots of the differential expression proteins were analyzed by Matrix-Assisted LaserDesorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS).For the further examinaton of regarding protein expression relations, The differential expression ofproteins were analyzed by bioinformatics and RT-qPCR respectively.Electrophoretogram of placenta tissues from cows with RFM and health control were obtained by2D-DIGE. A total of240protein spots (212up-and28down-regulated) were differentitallyexpressed in foetal placenta tissue of dairy cow,214(134up-and80down-regulated) differentialexpression protein points were founded in placenta maternal tissue of dairy cow.Ten differential expressed proteins were separated and identified by MALDI-TOF-MS and thesearch of NCBI database. Compared with the health control, in foetal placenta tissue, theupregulated proteins in RFM group were apolipoprotein A-I, annexin A8-like1, serine (or cysteine)proteinase inhibitor, glutathione transferase and transketolase,and the upregulated proteins in RFMgroup was BSA and alpha enolase; in placenta maternal tissue, the upregulated proteins in RFMgroup were serine (or cysteine) proteinase inhibitor, alpha enolase and annexin A2, and theupregulated proteins in RFM group were aldose reductase and heat shock27kDa protein1. Thedifferential expression of proteins in placenta of the cow with RFM were analyzed in the online analysis of GO, and Pathway. The cause and influencing factors of this disease is related tofibrinolysis, pyruvate metabolism, inflammatory response and oxidative stress.The results of the RT-qPCR was that: compared with the health control, in Foetal placenta tissue,The expression of apolipoprotein A-I, serine (or cysteine) proteinase inhibitor, glutathionetransferase mRNA were significantly higher (p<0.01).The expression of annexin A8-like1mRNAwas higher (p<0.05),The expression of transketolase was lower (p<0.05).In Placenta maternaltissue, The expression of serine (or cysteine) proteinase inhibitor and annexin A2mRNA weresignificantly higher (p<0.01), The expression of heat shock27kDa protein1mRNA weresignificantly lower (p<0.01).The identified proteins may be involved in the development of retained fetal membranes and stillneed further validation. The cause and influencing factors of this disease is related to fibrinolysis,pyruvate metabolism, inflammatory response and oxidative stress. However, it need to verify theseproteins and their relationship with RFM in the future, and offer a new strategy to interpretmechanism of RFM and to assess state of an illness and effect of prevention and cure in future.