| Objective The endometriosis(EMT)rat model and the postoperative recurrence state model were established,and the rats with EMT recurrence state model were treated with the comprehensive scheme of jianpishugan qingshihuayu(Traditional Chinese medicine intragastric administration + rectum introduction of Neiyi rehabilitation Suppository).By observing the general situation of rats,ectopic cyst formation and size,extent of pelvic adhesions and uterine residual oedema circumstance,the model rats in endometrium and ectopic lesions and pathological observation analysis of appearance,as well as the detection of Th1/Th2-related factor IFN-?,the expression of IL-4 after treatment and the relationship between them,to explore the immune mechanism of the comprehensive treatment of jianpishugan qingshihuayu for postoperative recurrence of EMT.Methods The EMT rat model was established by autologous endometrial transplantation.1 month later,partial ectopic lesions were resected to establish the recurrence state model after EMT.Then the rats in the model state of recurrence after EMT were randomly divided into the model group,the Gn group,the medium and high dose groups.The sham operation group was set up.There were 6 rats in each group.The model group and the sham operation group were given normal saline and rectal instillation,and the other groups were given different drugs for 4 weeks of continuous instillation and rectal instillation.The model group and the sham operation group were given normal saline by gavage and rectum,and the other groups were given different drugs to intervene in the same way for 4 weeks.After the last administration,rats in each group were put to death,and the healthy uterus and ectopic lesions were removed by laparotomy.HE staining was used to observe the tissue morphology of the uterus and the heterotopic lesion in rats,and the expression of IFN-?and IL-4 in the uterine and heterotopic lesion markers of rats was detected by Elisa.During the experiment,the general conditions of the rats were observed,and the formation and size of ectopic cysts,pelvic adhesions and uterine edema of the rats were observed and recorded during the second and third operations(establishment of the recurrence state model after EMT and the execution of specimens from the rats).Result(1)General conditions of rats:(1)Weight: Both before and after treatment,Compared with the sham group,there was no significant difference in the treatment precursor weight of all rats with postoperative recurrence state(P>0.05).Compared with the model group of postoperative recurrence status,there was no significant difference in body weight between the treatment groups(P>0.05).Comparison of rats with postoperative recurrence state model before and after treatment: compared with rats with postoperative recurrence state model before treatment,the weight of rats in each group increased after treatment,which was statistically significant(P<0.05).(2)Behavior and state performance: After 2 months of observation and recording,all the rats showed no abnormal mental state and reaction,and only after EMT model and the model of postoperative recurrence state after EMT modeling,the rats showed low spirit,decreased activity and slow reaction within 2 to3 days.In addition,the feces in the middle and high dose groups were often thin and mushy during gavage and enema.Gn group's fur was sparse and disorderly,and the activity was reduced.(2)Formation of ectopic cyst:(1)EMT model rats: The formation of heterotopic lesions such as ectopic vessels,nodules or vesicles of different sizes was observed by naked eye.(2)Before treatment: There was no significant difference in the size of left and right ectopic cysts in the model rats before treatment(P>0.05).(3)After treatment:After treatment: Left ectopic cyst: Compared with the recurrent state model group,the ectopic cyst was significantly reduced in the high-dose group of traditional Chinese medicine(P<0.05),while the Gn group and the medium-dose group showed a trend of reduction(P>0.05).Right ectopic cyst: compared with the recurrent state model group,the Gn group and the TCM ectopic cyst were significantly reduced(P<0.01 and P<0.05).There was a tendency to reduce the size of ectopic cyst in the medium dose group,but the difference was not statistically significant(P>0.05).(4)Comparison of postoperative recurrence status model rats before and after treatment:Left and right ectopic cysts: The recurrence status model group and the Chinese medicine medium dose group showed a tendency of reduction before and after treatment(P>0.05).After treatment,the size of ectopic cyst in the high-dose group and the Gn group was significantly reduced,and the difference was significant compared with that before treatment(P<0.01 in the right Gn group and P<0.05 in the rest).(3)Pelvic adhesions and uterine stump edema:(1)Pelvic adhesions: before treatment: Compared with the sham operation group,there was no statistically significant difference between the other groups(P>0.05).There was no significant difference between the other groups(P>0.05).After treatment: compared with the sham operation group,there was no significant difference in the degree of pelvic adhesions between the other groups(P>0.05).Compared with the model group,there was no significant difference in pelvic adhesions between the treatment groups(P>0.05).Comparison of rats with postoperative recurrence state model before and after treatment: there was no significant difference in pelvic adhesions between rats with postoperative recurrence state model and rats with sham operation group before and after treatment(P>0.05).(2)Residual edema: before treatment:When the rat model of recurrence status after EMT was established,21 rats were recorded with residual edema,and the rate of residual edema was 70%(21/30).None of the 6 patients in the sham operation group had uterine stump edema.After treatment: 16(16/30)rats with postoperative recurrent status had uterine stump edema,and the rate of uterine stump edema was 53.3%.None of the 6 patients underwent sham operation had uterine stump edema.Comparison before and after treatment: except for the sham operation group and western medicine group,stump edema remained unchanged.In the medium dose group,stump edema was reduced from 6(20.0%)to 3(10.0%),and in the high dose group,edema was reduced from 5(17.0%)to 1(3.3%),which was less than that before treatment.The model group increased from 4(13.3%)to 6(20%).(4)Pathological manifestations of endometrium and ectopic lesions in model rats after EMT:(1)Endometrium: Normal uterine morphology.(2)Ectopic lesions: Recurrent state model group: Cyst-like structure was observed,the inner layers of the epithelium were monolayer columnar with regular arrangement,some subnuclear vacuoles were seen,similar to the morphology and structure of the endometrium,and vasodilatation,neovascularization and hyperplasia were observed in the interstitium.Compared with the model group,the proliferation of new capillaries was reduced in the medium dose group,the western medicine control group and the high dose group.Endometrioid epithelial cells and mesenchymal cells proliferated significantly in the medium dose group,followed by the western drug control group,and the lighter in the high dose group.(5)The expression of INF-?in model rats with postoperative recurrence state of EMT:(1)Endometrium:Compared with the sham operation group,the INF-?concentration in the endometrial tissue of rats in the postoperative recurrence status model group was significantly reduced(P<0.01).Compared with the model group of postoperative recurrence status,INF-? concentration in the endometrium of rats in the high-dose group and Gn group increased significantly(P<0.05),while INF-?concentration in the endometrium of rats in the middle-dose group increased significantly(P>0.05).(2)Ectopic lesions: Compared with the model group of postoperative recurrence status,INF-?concentration in ectopic endometrial lesions increased significantly in the high-dose group(P<0.05)and Gn group(P<0.01).There was a trend of increasing INF-?in ectopic endometrial lesions of rats in the medium dose group(P>0.05).(6)The expression of IL-4 in model rats with postoperative recurrence state of EMT:(1)Endometrium:Compared with the sham operation group,the IL-4concentration in the endometrium of rats in the model group was significantly increased(P<0.01).Compared with the model group,the concentration of IL-4 in the endometrium of rats in the western medicine control group was significantly reduced(P<0.01).The concentration of IL-4 in the endometrium of rats in the high-dose group and the medium-dose group showed a decreasing trend(P>0.05).(2)Ectopic lesions:Compared with the model group,the concentration of IL-4 in heterotopic lesions in the western medicine control group was significantly reduced(P<0.01).The concentration of IL-4 in heterotopic lesions of rats in the high-dose group and the medium-dose group showed a decreasing trend(P>0.05).Conclusion1.In this study,the EMT rat model was established by autologous endometrial transplantation.The ectopic endometrium of the model group was similar to that of the endometrium,with more new blood vessels forming in the stroma and a large number of stromal cells proliferating,indicating the success of the model.2.The results of this study showed that the expression of Th1/Th2-related cytokines INF-?and IL-4 was different between the model group and the treatment group in the recurrence state,suggesting that postoperative recurrence of endometrial dysplasia might be related to the immune imbalance of Th1/Th2,which was mainly manifested as the decreased INF-?expression and the increased IL-4 expression,and the increased INF-?expression and the decreased IL-4 expression in the treatment group after intervention.3.The results showed that the comprehensive plan of jianpishugan qingshi huayu method could regulate the immune balance of Th1/Th2 by regulating the expression of IFN-?and IL-4,which might be one of the mechanisms to prevent and control postoperative recurrence of EMT. |
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