| Mycoplasma hyopneumoniae (Mhp) is the main causative agent of Mycoplasma pneumonia of swine (MPS), which can cause pneumonic lesions by specific adhesion in the respiratory tract inducing necrosis of epithelial cells, causing. In this study explored the relationship between Mhp infection and host cell response processes in pigs, analyze the infection from two part of different protein and oxidative damage by porcine respiratory epithelial cells in response to the strength strains of Mycoplasma hyopneumoniae infection.Detection of oxidative damage of the infected tracheal epithelial cells in vitro, from pigs infected by Mhp strains168(Virulent strains) and168L (attenuated strain) was carried out in this study. The time and dose of infection, and nitrogen monoxide (NO) release of the infected cell was measured. On this basis the effects of hydrogen peroxide (H2O2) and the indirect immunofluorescence(IFA)and4’,6-diamidino-2-phenylindole (DAPI) was verified by assessing the differences in cell oxidative damage. The results indicate that the optimum infection time is36h post-infection and the infectious dosage is5×106CCU. The NO in the supernatants of the infected cells was significantly higher in the cells infected by the virulent strain as compared to those infected with the attenuated strain (P<0.05),while there is a significant difference between the groups inoculated with the virulent and attenuated strains against the control group (P＜0.01).Detection of H2O2shows that the oxidative damage to cells was significantly higher in the groups inoculated with the virulent and attenuated strain than that of the control group (P <0.01). Additionally, the results demonstrates a significant difference (P<0.05) in oxidative damage between the cells infected with the virulent against those infected with the attenuated strain. IFA results reveals that the cells inoculated with the virulent have higher quantities of cell adhesion than those infected with the attenuated strain while the DAPI results show that the cells infected with the virulent strain are more adhered than that of the group infected with the attenuated strain and the extent of damage to cells is also larger. Currently, a research study on proteomics focused on Mhp itself with few aspects regarding the host response to the respiratory pathogens, In this experimental study, the strength of strain168of Mhp was evaluated after the in vitro infection of pig tracheal epithelial cells (STEC) and then the whole proteins were separated by two-dimensional electrophoresis (2-DE). Using PDQuestV8.0software, the proteins were analyzed and compared on the basis of their expression of its mass spectrometry (T-test, difference>1.5) to give a total of16points difference in the expression and11proteins were successfully identified. From these differences protein spots,it also found that five protein spots were up-regulated, while four protein spots were down-regulated in\Mhp168infected cells protein map and one protein spots were up-regulated, while five protein spots were down-regulated in\Mhp168L infected cells protein map. Simultaneously in Mhp168and Mhp168L infected cells protein map,there were one up-regulated and three protein spots down-regulated.To verify the results of2-D gel electrophoresis, glyceraldehyde3-phosphate dehydrogenase (GAPDH) was chosen and subjected to Western blot analysis. The changes in protein abundance noted in the Western blot analysis were considerably consistent with the result obtained by proteomics, which confirms the accuracy of proteomic study.By analyzing the differences in oxidative damage and the differentially expressed proteins of the cells infected with either the virulent or the attenuated strains, the present study will help in understanding Mhp pathogenesis and thereby provide a reference for early diagnosis, vaccine development and the basis to explore the appropriate drug to be administered in order to manage porcine respiratory illnesses in swine. |
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