| Objective: 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD), which belongs to polychlorinated dibenzo-dioxins , is one of the most toxic compounds .It could enter into human body threatening our health through many ways. Endometriosis is one of common diseases of gynaecology, characterizing as pelvic pain, abnormal menstruation, infertility and mostly affecting the quality of life of patients. In recent years, many studies indicate that it has close relations between TCDD and EMs. Researches find TCDD exposing not only could result in the happening of EMs, but also accelerate EMs'development by disturbing internal environment of human. However, its mechanism is not very clear. EMs is the diseases dependent on estrogen, which is very important to EMs. Cytochrome aromatase P450 is the limiting velocity enzymes in the estrogen'composing. So, we guess one of the ways of TCDD affecting the development of EMs is that it promotes the composing of estrogen.Our experiment exposes mouse of B6C3F1 to certain doses of TCDD before or (and) their born and build mouse model. Then do statistics of survival rate of the endometrial focus and detect the expression levels of P450arom in the eutopic and ectopic endometrium to supply the Experimental and theoretical basis on the pathogenesis of EMs.Methods:Mouse model were built and exposed TCDD before or(and) after born. Survival rate of ectopic focus was observed and made statistics; Reverse transcription polymerase chain reaction and Immunohistochemical techniques were used to detect the expression of P450arom in eutopic and ectopic endometrium before and after molding. Then their relations were discussed in the occurrence and development of EMs.Statistical analysis was performed using SPSS17.0 software package. The result was shown by mean±standard deviation. Survival rate of ectopic focus was compared by Chi-square test,αwas adjusted 0.0125 and 0.0045 when within and without group were compared; the expression of P450arom within and without groups were compared by One-way ANOVA (homogeneity of variance) and non-parametric test(Heterogeneity of variance). P<0.05 was considered significant for statistical analysis.Results:1 the total survival rate (the rate of three ectopic foci at least one place is survival)of subcutaneous tissue ,peritoneum and ovary of mice is 100%(60/60). The survival rate of the ectopic focus in the ovary is highest and the peritoneum and subcutaneous tissue take the second place, 53%(32/60)and 35%(21/60).The simultaneously survival rate of three location is 20%(12/60). Chi-square test show that the survival rate of ectopic focus in the three location is different(X~2=29.187,P=0.000<0.0125);the survival rate of ectopic focus in subcutaneous tissue and peritoneum is different compared with ovary(X~2=29.008,P=0.000<0.0125;X~2=12.478,P=0.000<0.0125); the survival rate of ectopic focus in subcutaneous tissue is not considered significant compared with peritoneum (X~2=4.089,P=0.043>0.0125). The survival rate of 3-3 group is considered significant compared with control group in the peritoneal ectopic focus (X~2=8.224,P=0.004<0.00625).The survival rate of the rest groups are not considered significant ;the suvival rate of subcutaneous tissue is not different compared with that of peritoneum among the five groups(ectopic focus in subcutaneous tissue :X~2=5.421,P=0.247>0.0045;ectopic focus in ovary:X~2=9.673,P=0.058>0.0045).2 The expression of P450arom mRNA in the eutopic and ectopic endometrium among five groups before and after molding.2.1 The expression of P450arom mRNA in the eutopic endometrium among five groups before molding.The result of one-way ANOVA shows, the expression of P450arom mRNA is considered significant(P=0.000). The expression of P450arom mRNA increases gradually according to the order of five groups— 0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.0495±0.0031), 3-0(0.0612±0.079), 0-3(0.0814±0.073), 3-3(0.0926±0.042), 3-10(0.1016±0.0118). According to the result of SNK and the level of a=0.05, it is considered significant between every two groups (P<0.05).2.2 The expression of P450arom mRNA in the eutopic endometrium among five groups after molding.The result of one-way ANOVA shows, the expression of P450arom mRNA is considered significant (P=0.000). The expression of P450arom mRNA increases gradually according to the order of five groups: 0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.0902±0.0136), 3-0(0.1695±0.0072), 0-3(0.2367±0.0211), 3-3(0.2986±0.0139), 3-10(0.3644±0.0112). According to the result of SNK and the level of a=0.05, it is considered significant between every two groups (P<0.05).2.3 The expression of P450arom mRNA in the ectopic endometrium among five groups after molding.2.3.1 The expression of P450arom mRNA in the ectopic endometrium in subcutaneous tissue among five groups after molding.The result of non-parametric test shows, the expression of P450arom mRNA is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom mRNA increases gradually according to the order of five groups: 0-0,3-0,0-3,3-3 3-10. Their average expression is 0-0(0.1168±0.0013), 3-0(0.1980±0.0010), 0-3(0.2806±0.0048), 3-3(0.3873±0.0021), 3-10(0.4817±0.0038). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups(P<0.05).2.3.2 The expression of P450arom mRNA in the ectopic endometrium in peritoneum among five groups after molding.The result of non-parametric test shows, the expression of P450arom mRNA is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom mRNA increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-10. Their average expression is 0-0(0.1236±0.0009), 3-0(0.2269±0.0011), 0-3(0.3390±0.0043), 3-3(0.4345±0.0037), 3-10(0.5009±0.0016). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups(P<0.05).2.3.3 The expression of P450arom mRNA in the ectopic endometrium in ovary among five groups after molding.The result of non-parametric test shown, the expression of P450arom mRNA is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom mRNA increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.1240±0.0014), 3-0(0.3277±0.0012), 0-3(0.4507±0.0015), 3-3(0.5646±0.0010), 3-10(0.6355±0.0011). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups(P<0.05).3 The expression of P450arom protein in the eutopic and ectopic endometrium among five groups before and after molding.3.1 The expression of P450arom protein in the eutopic endometrium among five groups before molding.P450arom protein is not expressed in the group of 0-0 before molding and is expressed at the rest four groups. The result of one-way ANOVA shows, the expression of P450arom protein is considered significant (P=0.000). The expression of P450arom protein increases gradually according to the order of four groups—3-0,0-3,3-3,3-10. Their average expression is 3-0(0.1464±0.0338), 0-3(0.1756±0.0472), 3-3(0.2345±0.0546), 3-10(0.2794±0.0330). According to the result of SNK and the level of a=0.05, it is considered significant between every two groups (P<0.05).3.2 The expression of P450arom protein in the eutopic endometrium among five groups after molding.P450arom protein is all expressed in five groups after molding. The result of one-way ANOVA shows, the expression of P450arom protein is considered significant (P=0.000). The expression of P450arom protein increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.2084±0.0215), 3-0(0.2355±0.0248), 0-3(0.2663±0.0309), 3-3(0.2907±0.0353), 3-10(0.3173±0.0270). According to the result of SNK and the level of a=0.05, it is considered significant between every two groups (P<0.05).3.3 The expression of P450arom protein in the ectopic endometrium among five groups after molding.3.3.1 The expression of P450arom protein in the ectopic endometrium among five groups in subcutaneous tissue after molding.P450arom protein is all expressed in five groups in ectopic focus of subcutaneous tissue after molding .The result of non-parametric test shows , the expression of P450arom protein is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom protein increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.2412±0.0136), 3-0(0.2664±0.0223), 0-3(0.2985±0.0333), 3-3(0.3509±0.0014), 3-10(0.4092±0.0017). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups (P<0.05).3.3.2 The expression of P450arom protein in the ectopic endometrium of peritoneum among five groups after molding.P450arom protein is all expressed in five groups in ectopic focus of peritoneum after molding .The result of non-parametric test shows , the expression of P450arom protein is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom protein increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-10. Their average expression is 0-0(0.2597±0.0014), 3-0(0.2988±0.0190), 0-3(0.3286±0.0220), 3-3(0.4425±0.0288), 3-10(0.5202±0.0115). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups (P<0.05). 3.3.2 The expression of P450arom protein in the ectopic endometrium of ovary among five groups after molding.P450arom protein is all expressed in five groups in ectopic focus of ovary after molding .The result of non-parametric test shows , the expression of P450arom protein is considered significant in the ectopic endometrium among five groups(P=0.000). The expression of P450arom protein increases gradually according to the order of five groups—0-0,3-0,0-3,3-3 3-3. Their average expression is 0-0(0.2853±0.0219), 3-0(0.3381±0.0347), 0-3(0.4337±0.0514), 3-3(0.5409±0.0551), 3-10(0.6337±0.0293). According to the result of two independent sample non-parameter test and the level of a=0.05, it is considered significant between every two groups(P<0.05).4 The expression of P450arom mRNA and protein in proliferative phase (preoestrus and estrum) and secretory phase(postestrus and anoestrum) is not considered significant before and after molding (P>0.05)Conclusion:1 The survival rate of ectopic focus in ovary is higher than that in subtaneous tissue and peritoneum in the mice models.2 Regular doses of TCDD could boost the success of ectopic focus in the mice models.3 The expression of P450arom is higher and higher in the eutopic endometrium in mice models when the dose and time of TCDD exposing is increasing before molding, showing that TCDD could boost the EMs'attack by increasing the expression of P450arom.4 The expression of P450arom is higher and higher in the eutopic endometrium in mice models when the dose and time of TCDD exposing is increasing after molding; its expression is higher after molding than before molding, showing that TCDD could produce new ectopic focus by expressing more P450arom in eutopic endometrium and worsing the environment in the eutopic endometrium.5 The expression of P450arom is higher and higher in the ectopic endometrium in subcutaneous tissue, peritoneum and ovary in mice models when the dose and time of TCDD exposing is increasing after molding; its expression is higher after molding than before molding, showing that TCDD could deteriorate EMs by expressing more P450arom in ectopic focus.6 The expression of P450arom is higher in the ectopic endometrium in ovary than subcutaneous tissue and peritoneum, showing that pathological damage in ovary is heavier than that in subcutaneous tissue and peritoneum.7 It could induce the generation and development of EMs by TCDD exposing before or(and) after born. The rate of producing EMs is higher and the illness is more serous by TCDD exposing before and after born than by before (or) after born.
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