The Effects Of Endostatin Gene On Ang-2,Tie-2 And MVD In The Transplanted Model Of Endometiosis In Babl/c Mice

Objective:By using experimental zoology,pathology and immunology methods,Our study intents to establish the nude mouse endometriosis model,injecting in local focus with recombination adenovirus to investigate the efftcts of endostatin to Ang-2,Tie-2 and MVD, in order to find out a new therapy way for endometriosis which is convenient and safe.Our experiment contains two parts:Part one:Establishment of nude mouse endometriosis modelObjects:Through the construction of nude endometriosis model to explore the feasibility of establishing nude mouse model of EMS and specific methods.Methods:Using peritoneal seeding method, to establish EMS model in nude mice by removing the person eutopic endometrium into nude mice. after 7 days of transplantation, observe the abdominaland pelvic lesion of and abdominal by naked eye, containing:size,appearance and growth of blood vessel. and to take cases of lesions for HE examination, to understand the success of animal models.Results:Nude mice EMS model was established successfully, endometriotic lesions grew well, showing a small cystic bulge translucent,and clearly visible blood vessels. By microscopic observation, clearly endometrial glands and stroma can be seen,the neovascularization of surrounding stromal is very rich. Animal model is established successfully.Part two:The effects of endostatin on Ang-2/Tie-2 system in the transplanted model of endometiosis in Babl/c miceObjects:To investigate the effects of endostatin on Ang-2/Tie-2 and MVD system in ectopic lesion of EMS model.Methods:Model mice were divided into three groups randomly including rAAV2-hEndostatin-EGFP group(n=20), rAAV2-EGFP group (n=20)and PBS control group(n=20).Histology changes were examined among the three groups,and the expression of Ang-2\Tie-2 and MVD were examined by immunohistochemistry. Results:(1) The gland proliferation of the two control groups is obvious,the mesenchymal blood vessel is rich;but the gland of endostatin group withers,the blood vessel reduces either.; The number of glands and microvessel density in indostatin group were significantly less than the empty virus group and the PBS control group. (2) The expression of MVD in endostatin group is lower than other two groups,which also has statisticly significant difference. (3) The expression of Ang-2 and Tie-2 is mainly in the glandular epithelium,and less in vascular endothelium and stroma locating in cytoplasm.The positive rate of expression about Ang-2 in endostatin group was 40%,much lower than that in other two groups(75%, 80%P< 0.05).The positive rate of expression about Tie-2 in endostatin group was 45%,also lower than that in other two groups(80%,85%P< 0.05).Conclusion:(1)We successfully established endometriosis animal model in nude mice by using human dendometrioal tissue.Model traits appeared stable and retained the original characteristics of the organization.(2) After injuction of endostatin,the expression of Ang-2 and Tie-2 were inhibited significantly and MVD were also reduced which could effect the angiogenesis, leading to gland atrophy and reduction in the number for therapeutic purposes.