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Effects of Methoxychlor on Uterine Toxicity and ANGPT-1 Expression of Endometrium

Author: DuJuan
Tutor: ChenBiLiang；ZhangJianFang
School: Fourth Military Medical University
Course: Obstetrics and Gynaecology
Keywords: Environmental endocrine disruptors Methoxychlor Endometrial Angiopoietin-1 Immunohistochemistry Reverse transcription-polymerase chain reaction
CLC: R114
Type: Master's thesis
Year: 2009
Downloads: 47
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Abstract

Environmental endocrine disruptors (Endocrine Disrupting Chemicals, EDCs) is defined as the hormone of the body to maintain its own balance, reproductive, developmental and behavioral synthesis, transfer, bonding, role, or clear the interference effect of exogenous substances, these substances can analog, enhance or inhibit hormone synthesis, in certain cases, may lead to a tissue or organ dysfunction and tumorigenesis. More and more human exposure to this type of interference was due to occupational and environmental factors seriously affect the human reproductive function and good health. Methoxychlor as environmental endocrine disruptors one (methoxychlor, MXC), in recent years, has been widely applied, instead of the organochlorine pesticides DDT, and DDT compared with mammals metabolism fast, easy excretion, toxicity and low. MXC and its metabolites have been shown to cause female rat hypothalamic - pituitary - gonadal axis ovarian disorder, synthesis and secretion of steroid hormones to reduce endometrial thickening, increased quality, cyclical changes may affect the follicle. MXC on endometrial angiogenesis currently rarely reported. Research methoxychlor on uterine tissue, especially endometrial vascular generated toxic effect is of great significance. This study by constructing female mice methoxychlor exposure model to observe the expression of angiopoietin-1 on the female endometrium to further explore the role MXC development of endometrial disease, trying for clinical diagnosis and The treatment provides new theoretical basis. This experiment is divided into three parts: The first part: the model of female mice exposed to methoxychlor and its toxic effects on the uterus of 40 adult female Kunming mice adaptive feeding for 7 days, were randomly divided into solvent sesame oil (control group 21d consecutive days after intraperitoneal injection MXC exposure, the mice were killed immediately after exposure, the observed indicators are: body weight, uterine proliferation index) and MXC 50, 100, 200 mg / (kg · d) group; hematoxylin - eosin staining (Hematoxylin- Eosin Stain, HE) light microscope to measure the height of the proliferation of endometrial epithelial cell layer and stromal cell layer thickness; measured by radioimmunoassay in serum E2 and P levels. Results: ① The weight of each dose group into a growth trend, but no significant difference in the high dose group endometrial epithelial cell proliferation height and stromal cell layer thickness with the control group compared to significant differences. ② in the high dose group uterine proliferation index with increasing dose and elevated; ③ The serum E2 levels in the high dose group significantly reduced P with the control group showed no significant difference. Conclusion: ① The MXC can increase uterine weight increase endometrial epithelial cell proliferation height and stromal cell layer thickness; ② in high dose MXC possible through under the influence of hypothalamic - pituitary - ovarian axis and thus make E2 levels reduce; ③ The low-dose group of uterine tissue No clear impact. Part II: methoxychlor mouse endometrium angiopoietin-1 protein expression levels exposed to experimental one paraffin-embedded uterine tissue specimens observed indicators have: SP immunohistochemistry and immunofluorescence under the microscope, calculate the average optical density of each dose group; immunohistochemistry SP method endometrial tissue observed in each dose group ANGPT-1 protein levels. Results: ① SP of increasing exposure dose was significantly higher in the high dose group ANGPT-1 protein expression in the endometrium, the average optical density of the lower-dose group and the control group increased, the low-dose group and the control group had no significant difference; ② compared to secretory phase endometrium in the high dose group ANGPT-1 increase the expression in the proliferative phase is more evident. Conclusion: ① in the high-dose MXC can increase the expression of endometrial tissue in mice angiopoietin ANGPT-1 protein; ② in the high dose group endometrial ANGPT-1 in the proliferation of the increase may be was estrogen dependent, may promote uterine endometrial repair, also may contribute to abnormal endometrial angiogenesis, such as endometrial hyperplasia and endometrial cancer, and so on; may be an increase in the secretion of the VSMC proliferation and to promote the spiral arteries of mature and stable. Part III: methoxychlor exposure on the the mouse endometrium the angiopoietin-1mRNA expression level of the impact animal exposed to the ways and means with the experiment, the mice in each group were sacrificed quickly remove the endometrial tissue, RNA was extracted. The detection ANGPT-1 mRNA expression level using reverse transcription polymerase chain reaction (RT-PCR) method to calculate the relative content of ANGPT-1mRNA. Results: MXC in the high dose group ANGPT-1mRNA relative content was significantly higher than that in the control group, low dose group and the control group showed no differences. Conclusion: The high dose MXC can improve mouse endometrium ANGPT-1 gene transcription level, thereby affecting the physiological and pathological processes in the endometrium.

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