Analysis Of The Expression Of Endostatin Gene And Protein After Endostatin Gene Transferred Into Endometriosis Rat

Objective: To judge the result of ES gene transfection and to investigate the probability of the way using in the treatment of endometriosis from injecting the Hpofectamine-endo-pBud compounds to the ectopic focus of endometriosis rat model by Western-blot and real-time FQ-PCR.Methords: Eighty-five 14-week-old female unpregnancy Lewis rats deployed autotransplantation to construct endometriosis model. Evaluated the achievement ratio 4 weeks later. Select the sixty-four successful ones as empirical study objects.Divided them randomly into endostatin transgenic group(I group),carrying agent control group(II group) and negative control group(III group). Injected 2μl Hpofectamine-endo-pBud compounds using microinjector to ectopic focus of I group, 2μl lipofectamine-pBud compounds to II group and 2μl PBS to III group by the same way. After 2 weeks divided every group into 2 subgroup randomly. Taked the EF tissue to measure the expression of endostatin gene were by real-time fluorescent quantitative PCR , the expressions of ES-HA fusion protein and endostatin protein by Western-blot.Results:1. In the experimentation 64 rats among all of the 85 ones formed ectopic focus successfully. The achievement ratio is 75.29% for using autotransplantation to construct endometriosis rat model.2. Two weeks after injecting lipofectinmine-endo-pBud compounds to the EF directly, the expressions of endostatin gene were detected by real-time fluorescent quantitative PCR, found the difference between endostatin transgenic group and in carrying agent control group and negative control group has statistical significance (P < 0.05) ,but the two control groups did not reach statistical significance(P > 0.05);ES-HA fusion protein can be detected in the ectopic focus of endostatin transgenic group. The expression of endostatin protein is much higher in the ectopic focus of endostatin transgenic group than in carrying agent control group and negative control group(P < 0.01).Conclusion:1. The model of auto-transplanted surgically Lewis rats is feasibility and repeatability, it is a suitable endometriosis animal model.2. ES gene and protein can expressed in endometriosis rats successfully by injecting lipofectinmine-endo-pBud compounds to the EF directly.