| Objective To reveal the mechanism of endometriosis and its recurrence and to demonstrate the effect of Luoshi Neiyifang(LSNYF), endometriosis models and recurrence models in rats were set up first. The network of estrogen regulation in local ectopic endometrium of endometriosis were explored. Comparison with Danazol, the effects of LSNYF on estrogen regulation in local ectopic endometrium of endometriosis were observed. The functions of LSNYF to recurrence models were detected at the same time.Methods Self-endometrium transplantation by surgical method was used to develop rats models of endometriosis. After 7 weeks the recurrence ones were built by removing partial ectopic endometrium. Compared with blank group and oophorectomized group, they had been raised by LSNYF(high concentration and low concentration), Danazol, Diethylstilbestrolum(DES). During that 7 weeks, the rats in group A had been raised by different mentioned medicine for 6 weeks and ectopic endometrium was excised partially in the end of the third week. In group B, no treatments were carried out in the first 3 weeks before the partial excision. Ectopic endometriosis was observed both by macroscopic and microscopic analysis. Radioimmunoassay was used to determine the level of E2 and ELISA was used in PGE2 test in ectopic endometrium.Fluorescence quantitative PCR(FQ-PCR) was used to detect the expression of C-FOS, P450arom, SF-1, COUP-TF in ectopic endometriums.Results (1)The rats with endometriosis were normal in activity, coat color appetite and excreta. The rats with recurrence lesions were normal after the excision. The death rate after excision was not related to the volume of ectopic endometrium and adhesion degree(P>0.05).(2) The recurrence ectopic lesions with rich vessels were varied morphology. Even though no cysts can be seen in the recurrence area, nodules and scars can be found with rich vessels. All the recurrence lesions appeared individually and no disseminated ones can be found. The endometrial epithelium and stromal was confirmed in the recurrence ectopic lesion, which meant that the establishment of endometriosis recurrence was succeed.(3)The statue and activities with different treatments were follow: (â…°)The rats in blank group were normal in activity, coat color appetite and excreta, (â…±)There were no significant difference with weight in other treatments. The behaviors were less active with blank group. But they recovered soon after the excision except the DES group. (â…²)In DES group, the weight was the least (p<0.05) in all groups. They were flagging, slow, poor appetite with mashy stools.(4)The results before the excision are following (group A):(â…°)The contents of E2 and PGE2 in ectopic endometrium in group of LSNYF with high concentration(LSNYF-H), LSNYF with low concentration(LSNYF-L),Danazol were lower significantly than blank one (p<0.05), while the DES group was higher sinificantly (p<0.05). There were no significant difference between LSNYF-H, LSNYF-L and Danazol group (p>0.05). (â…±)The expression of C-FOS in ectopic endometrium in group of LSNYF-H, LSNYF-L, Danazol were lower significantly than the blank one (p<0.05). There were no significant difference in C-FOS between LSNYF-H, LSNYF-L and Danazol group (p>0.05). (â…²) The expression of P450arom in ectopic endometrium in group of LSNYF-H, LSNYF-L, Danazol were lower significantly than the blank one (p<0.05) while the DES group was higher sinificantly (p<0.05). The LSNYF-L group was also lowest in P450arom expression (p<0.05). (â…³) The expression of SF-1 in ectopic endometrium in LSNYF-L was lower significantly than the blank one (p<0.05) while the DES group was higher sinificantly (p<0.05). The LSNYF-L group was also lower significantly than LSNYF-H and Danazol groups (p<0.05) in SF-1 expressions. (â…´) There were no significant difference between every group in COUP-TF expressions (p>0.05). (â…µ) The E2 content was positively correlated with PGE2, P450arom, C-FOS(p<0.05) and negatively correlated with COUP-TF(p<0.05).PGE2 had positive correlation with P450arom expression (p<0.05). C-FOS expression had positive correlation with COUP-TF and E2(p<0.05). P450arom expression had positive correlation with SF-1, E2 and PGE2. There were no relationship between COUP-TF and SF-1 (p>0.05). (â…¶) The volume of ectopic endometrium in LSNYF-L was the smallest (p<0.05) in all groups while the DES'was the highest (p<0.05). There were no significant difference between LSNYF-H, Danazol, oophorectomized and blank group (p>0.05). The adhesion degree of LSNYF-H and LSNYF-L was the most mildly (p<0.05).(5)The results after excision of ectopic endometrium are following:(â…°) In group A, the contents of E2 and PGE2 in all treatment groups were lower than the blank one significantly(p<0.05) while the DES group was the highest(p<0.05). (â…±)In group B, the contents of E2 in all treatment groups were lower than the blank one significantly(p<0.05) while the DES group was higher than LSNYF-H and Danazol significantly(p<0.05). There were no significant difference between LSNYF-H, LSNYF-L and Danazol group (p>0.05). The content of the DES group was highest in all groups (p<0.05). There were no significant difference between LSNYF-H, LSNYF-L, Danazol and the blank group (p>0.05). (â…²)The volume of the ectopic endometrium in LSNYF-L A was smaller than the blank group while the DES group was much bigger (p<0.05). The adhesion degree of LSNYF-L was the most mildly (p<0.05).(6)The difference between the endometriosis and recurrence endometriosis models are showing as following:(â…°)Except the oophorectomized, blank,and LSNYF-L B groups, the contents of E2 were lower after excision in other groups(p<0.05). The contents of PGE2 were lower after excision in LSNYF-L A, Danazol A, LSNYF-H B groups (p<0.05). (â…±) The adhesion degree of LSNYF-H A and B was improved significantly after excision (p<0.05). (â…²)The volumes of ectopic endometrium were reducing significantly after excision (p<0.05).Conclusion (1)Establishment of recurrence endometriosis model by autologous transplantation on SD rats was success and credible. The endometrial epithelium and stromal was confirmed in the recurrence ectopic lesion.(2)The recurrence ectopic lesions with rich vessels were varied morphology. Some appeared invasive with no cysts. The idea that the cysts are the observation index of success endometriosis models is not suitable. The study showed that the volume of ectopic endometrium, might be the index of effect of treatment if the ectopic endometrium cysts could be seen. But the adhesion degree cannot demonstrate the effect of treatment.(3)The study revealed that there were two independent ways of estrogen regulation in ectopic endometrium, the network of SF1-P450arom-PGE2 and the early response to estrogen of C-FOS, which caused the high concentration of E2 in ectopic endometrium and promoted the disease. The excision of ectopic endometrium can lessen the volume of the lesion and cannot reduce the content of E2 and PGE2. The result showed that the promotion of estrogen synthesis didn't change after excision of partial ectopic endometrium. That may be the reason of endometriosis recurrency.(4) LSNYF with certain concentration may reduce E2 in ectopic endometrium through inhibiting the expression of SF-1 and C-FOS. The cysts are lessen and the micro-environment is improve after treatment by LSNYF. The measure that is dealt with LSNYF-L before and after operation can reduce risk and prevent recurrency.
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