| Schistosomiasis is a extensive and serious hazard zoonotic parasitic disease.Praziquantel can kill adults and cercariae,which is an effective and safe therapeutic drug,but it cannot prevent infection.Objective factors for the prevalence and spread of schistosomiasis and risk factors for the recurrence of schistosomiasis still exist in some areas in china.New drugs and vaccines are key points of study on prevention and control technology of schistosomiasis.M.fortis is a mammal with natural resistance to S.japonicum,and the study of its resistance mechanism can provide new directions and clues for the prevention and control technology of schistosomiasis.In this paper,DMEM was used to lavage of the abdominal cavity of M.fortis,which was quickly shaken horizontally,then the macrophage was isolated,cultured and purified by adherence.Thus to establish a simple method for the isolation and culture of the peritoneal macrophages of Reed vole.As a result,a large number of high-purity peritoneal macrophages were obtained with normal morphology and about 98% viable cells.The cultured cells have a good phagocytic function confirmed by ink particles phagocytosis test.The staining results of intracellular acid phosphatase and non-specific ?-phenol naphthol esterase showed that the obtained cell contained abundant hydrolasesin the cytoplasm.The results indicated that the rapid shaking method is a simple and practical method for the isolation and cultivation of peritoneal macrophages from M.fortis.The NO content in the serum and liver and lung tissues of M.fortis and BALB/c before and after infection was measured.The results indicated that the NO content in M.fortis infected at 0d,3d,7d,and 15 d showed an upward trend,which were 3.99 ?M,4.95 ?M,18.89 ?M and 23.81 ?M respectively,and higher than those in BALB/c serum at the same timepoints.During infection,NO levels in the liver tissues of M.fortis and mice showed a downward trend,and the difference was not significant.The NO content in the lungs of M.fortis before and after infection 3d,7d,and 15 d increased and then decreased,which was significantly lower than that of BALB/c mice.The NO content produced by M.fortis macrophages was significantly higher than that produced by KM macrophages,and the NO content after LPS or IFN-? stimulation was significantly higher than that of unstimulated cells or KM macrophages.The results of fluorescent probe analysis showed that the proportion of M.fortis macrophages with NO activity was greater than that of KM macrophages.After the addition of LPS-stimulated macrophages and M.fortis serum for 24 h,a large number of cells adhered on the surface of larvae,which was significantly more than inactivated M.fortis serum and unstimulated macrophages.The schistomula killing effect of the normal sera of M.fortis and the LPS-stimulated macrophage group was significantly greater than that of the other groups.The schistosomicidal effect after adding macrophages was significantly different from that without macrophages.The results show that NO produced by macrophages and serum complement can synergistically kill schistosomula.In this paper,the Complement binding proteins rSjTOR-ed1 and rSjC1 qbp of S.japonicum were expressed and purified.The test results showed that the recombinant hemolytic inhibition rates of 10 ?M rSjTOR-ed1 and 5 ?M rSjC1 qbp recombinant proteins on M.fortis serums were 37.47% and 81.73% and on KM serums were 55.61% and 91.99%,respectively,the difference is very significant.The results indicated that the inhibitory effects of S.japonicum complement inhibitor proteins rSjTOR-ed1 and rSjC1 qbp on the serum complement activity of M.fortis were lower than those of KM,which provided data for explaining the role of complement in the mechanism of M.fortis against S.japonicum.The results of this paper provide data for understanding and elucidating the mechanism of M.fortis against S.japonicum. |
PDF Full Text Request