The Study On Proteome Of Liver Tissue Of The Porkets Experimentally Infected By Taenia Asiatica

Objective: In order to provide substantial scientific information for exploring the molecular mechanism of porcine liver injury caused by Taenia asiatica,proteomic methods were employed to research the porcine liver experimentally infected by T.asiatica.Methods: 1.The T.asiatica adults were collected from the taeniasis patients in Liangmu Township of Duyun City,Guizhou Province and identified biologically.The eggs were harvested from gravid proglottids and prepared by repeated washing and centrifugation.2.Twelve 20-days old Yorkshire and Seghers hybrid porkets were randomly divided into experimental and control groups as six pigs per group.The experimental group was orally administrated with 1.5×106 eggs per porket at day 0 post-infection.The porkets of both groups were sacrificed on the day 15 and day 75 post-infection(three pigs per time point)respectively,and liver samples were collected for further experiments.3.The unbiased iTRAQ proteomic method was employed to research the porcine liver experimentally infected by T.asiatica.The main steps included preparation of protein,enzymatic hydrolysis and peptide quantification,peptide labeling,liquid chromatography tandem mass spectrometry analysis,identification and quantitative analysis of Mascot software and gene ontology analysis.4.Quantitative real-time polymerase chain reaction,western blotting and immunohistochemistry methods were used to further verify some target proteins.Results: 1.According to the general morphological characteristics and molecular identification,the tapworms collected in Liangmu Township of Duyun City were T.asiatica.2.The infection rate of the experimental group was 100%.Cysticerci of T.asiatica were found only in the porcine liver.3.Compared with the control group,187 proteins were differentially expressed in infected livers at the day 15 post-infection,including 92 unknown proteins.Compared with the control group,158 proteins were differentially espressed in infected livers at the day 75 post-infection,including 72 unknown proteins.4.At the day 15 post-infection,the mRNA level and expression level of Caspase-3 of the experimental group were significantly decreased,comparison with the control group(P<0.05,P<0.01 respectively).However,at the day 75 post-infection,the mRNA level and expression level of Caspase-3 of the experimental group were dramatically similar to the control group.At the day 15 and 75 post-infection,the mRNA levels and expression levels of Anxa5 and Metap2 of the experimental group were all significantly increased,comparison with the control group(P<0.05,P<0.01 respectively),while the mRNA level and expression level of Cyp1a1 of the experimental group were significantly decreased,comparison with the control group(P<0.05,P<0.01 respectively).It was positive with Caspase-3 or Cyp1a1 when yellow or brown signal appeared in the cytoplasm of liver cells by immunohistochemistry.Conclusion: 1.Quantitative proteomic analysis results in porcine liver with iTRAQ technology provided substantial scientific information for further exploring the molecular mechanism of porcine liver injury caused by T.asiatica.2.The results for verification of target proteins suggested that Caspase-3 might be involved into the regulation of the damage of porcine liver at day 15 post-infection after oral challenge by T.asiatica and have an effect on the hepatic injury because of the dramatical recovery of Caspase-3 at the consequent infection stage.Meanwhile,Anxa5?Metap2 and Cyp1a1 might be involved into the regulation of the damage of porcine liver at day 15 and 75 post-infection after oral challenge by T.asiatica and have an effect on the hepatic injury.