Study Of The Warburg Effect Markers In A Modified Animal Model Of Endometriosis

I Construction of a subcutaneous model of endometriosis in immunodeficient nude miceObjective:To establish a model of human endometriosis in nude mice,and to analyze the modeling effect of two different treatments after subcutaneous transplantation.Materials and Methods:Endometrial tissues were obtained from the patients who met the criteria in our hospital.The endometrial tissues were subcutaneously implanted in standard nude mice(nude mice were provided by the Animal Centre of Sir Run Run Shaw Hospital,School of Medicine,Zhejiang University).Skin graft of one group was not sutured and that of the other group was sutured after transplantation.The success rate,the inflammatory response of skin wound and survival rate of models in two groups were calculated.The modeling effects of two different treatments of subcutaneous tissue blocks after transplantation on the incision were analyzed.Results:(1)The general situation:nude mice of the non-sutured incision group were alive,but one.of the sutured group died.The postoperative wound in the non-sutured group was completely healed after 3d.The external suture of the wound in the sutured group fell off and healed well after 2?4d.There was one mouse with no lesion in the D7 group of the non-sutured group,and one mouse in the sutured group died because of abdominal distention after 1 day.There was one mouse with no lesion observed in the D14 group of the sutured group.There was no significant difference in the time of lesion formation between the two groups.General observations found single spherical ball lesion after 7d,which then became more clear small vesicle-like protrusions in pale pink,lesions were surrounded by rich new blood vessels.14d later,lesions turned into single large bubble-like protrusions in pale pink,and peripheral blood vessels were reduced than before.After 21d,lesions turned to gray cystic protrusions,the lesion volume was smaller than that on 14d.(2).HE staining:typical glandular structure and interstitial tissue structure were seen after one week,two weeks and three weeks of operation,with glands in proliferative phase.Endometrial tissue before implantation served as DO group were detecte by IHC.The results showed that the endometrium were all in proliferative phase proliferated.(3)The growth rate of the lesions:after modeling group a,the cumulative rate of model formation and the survival rate of nude mice were 1/3 and 3/3 in one week,3/3 and 3/3 in two weeks,and 3/3 and 3/3 in three weeks.In group b,the cumulative rate of model formation and cumulative survival rate of nude mice were 2/3 and 2/3 in one week,2/3 and 3/3 in two weeks,and 3/3 and 3/3 in three weeks.The total success rate of modeling of group a was 8/9,group B was 7/9;survival rate of group a was 9/9,group B was 8/9.The results showed that the difference was not statistically significant,and no significant difference was found in wound healing between the two groups.Conclusion:(1)Subcutaneous transplantation is an ideal method,which offers more directly and more accurately to calculate the size and number of ectopic foci,and provide convenient.(2)There were no significant differences in the three indexes between the subcutaneous non-sutured group and the sutured group.However,didn't suture of the wound reduced the experimental steps and shortened the operation time,so it is worth to be popularized in the establish of mice model of endometriosis.? Study on the Expression of Warburg Effect Markers in the Animal Model of EndometriosisObjective:The Warburg effect(anaerobic glycolysis)were reported to be involved in the development of endometriosis.Using improved nude mouse model of human endometriosis,we delected the levels of hypoxia-induced glycometabolism-related markers(including HIF1?,VEGF,GLUT1,LDHA,PDKl).We aimed to clarify whether Warburg Effect participated in the pathogenesis of EM.Materials and Methods:Fourteen patients without endometriosis that met the inclusion criterias in our hospital were selected to establish the modified model of endometriosis.Then Quantitative Real-time PCR Detecting System and immunohistochemistry were performed to detect the expression of Warburg effect assaiated markers in ectopic lesions of mouse model.The expression levels of Warburg effect markers in one week,two weeks and three weeks after transplantation,including hypoxia-induced factor 1?(HIF1?),vascular endothelial growth factor(VEGF),glucose transporter(GLUT1),lactate dehydrogenase(LDHA),pyruvate dehydrogenase kinase(PDK1)were compared between different groups.Results:Eutopic endometrium scraped from nomal women were considered as the control group(the endometrial tissue was obtained directly from the female at the beginning of the experiment,denoted as 0d).Subcutaneous endometriotic tissues of the mice obtained after 1 day,7 days and 14 days of transplantation were taken as the study group(denoted as 1d,7d and 14d respectively).The expression levels of Warburg effect markers were detected by qRT-PCR and immunohistochemistry.QRT-PCR results showed that the expression of HIF-1?,VEGF,GLUT1,LDHA and PDK1 were significantly higher than that after 1 day or 7 days of transplantation.While on the D14,the expression of most genes returned to the DO level.Meanwhile,the results of immunohistochemistry found that the protein expression level of HIF-1?,VEGF,GLUT1,LDHA and PDK1 in glandular and interstitial cells on the D1 and D7 was also significantly higher than that on D0,and the expression on the D14 also returned to the D0.Conclusion:Our study confirmed that the Warburg effect in animal models of endometriosis marker at the beginning of the occurrence of endometriosis presented lower after the first high expression rule,prompt Warburg effect involved in the onset of vision.To clarify the different disease in metabolic pathway is helpful to the early onset of lesion of the etiology of the disease provide theoretical basis for treatment.