Effects Of Methoxychlor On Uterine Toxicity And ANGPT-1 Expression Of Endometrium

There are many exogenous chemicals which can interfere the synthesis, transportation, binding, function or elimination of hormones in vivo, which maitain balance, reproduction, development and behavior in human beings. These chemicals can mimic, strengthen or inhibiti the synthesis of hormones in vivo. Under some circumstances, they may result in abnormality of tissues and organs as well as tumor. These kinds of chemicals are defined as endocrine disrupting chemicals. More and more evidence indicated that the exposure of these chemicals through occupation and enviorment would influence and change the generation function of human beings.Methoxychlor(MXC) is an organochlorine pesticide, which is widely used to be an alternative to dichloro-diphenyl-trichloroethane (DDT) for the past few years for its less toxic and rapidly metabolized and excreted in mammals. It had been confirmed methoxychlor and her metabolism could disorder the hypothalamic-pituitary-ovarian axis, and then reduced synthesis and secretion of steroid hormone. They could increase the endometria thickness and qualities and might affect the cyclic variation of ovarian follicle. But the effects of methoxychlor on ANGPT-1 expression of endometrium in female mice had few reports at the present. For this reason, it is necessary to investigate the toxic effects of MXC on mice uterus, especially on ANGPT-1 expression of endometrium. The study was designed to analyze the effects of MXC on ANGPT-1 expression of endometrium through construction of methoxychlor poisoned mice model, which futher tried to expore the action of MXC in development and growth of pathological changes on uterus, tried to provide some useful theories for clinic diagnosis and treatment.Part 1. Construction of methoxychlor poisoned mice model and effects of methoxychlor on mice uterusMXC were administrated to forty female at doses of 0 (control), 50, 100, 200mg/(kg·d) via intraperitoneal injection for 21d, then mice were euthanized immediately. Body weights, proliferation index of uterus were observed;after Hematoxylin and eosin staining, measuring the hyperplastic hight of endometrium epithelial cell and thickness of endometrium stroma cell layer turn to light microscope; estradiol(E2) and progestereone(P) were detected with the radio immunoassays. Results:①Increasing of body weights was not different between 4 groups after treatment;the hyperplastic hight of endometrium epithelial cell and thickness of endometrium stroma cell layer in MXC 100 and 200mg/(kg·d) groups were obviously different from the control group.②Proliferation index of uterus were gradually increasing along with MXC 100 and 200mg/(kg·d) groups which were growing.③Estradiol (E2) were significantly decreased in 100, 200 MXC groups. There were no significant differences on serum progestereone (P) in 4 groups. Conclusion:①MXC could increased uterus weights and hyperplastic hight of endometrium epithelial cell and thickness of endometrium stroma cell layer.②MXC in experimental doses might through disorder the hypothalamic-pituitary-ovarian axis and induced the decrease of E2.③In low-dose groups there were no significant influence on uterus tissue.Part 2. Effects of methoxychlor on endometrium of ANGPT-1 proteinum expression in female miceWe used the uterus tissue embeded with paraffin just from the first experiment. Then the ANGPT-1 cell were observed by electron microscope; Calculating average optical in 4 groups; The ANGPT-1 proteinum expression in uterus were examined with the SP immunohisochemical method. Results:①The SP immunohisochemical method discovered the average optical in 100, 200mg/ (kg·d)groups was obviously higherthan that in 50mg/(kg·d)and control groups; The ANGPT-1 proteinum expression in 100, 200mg/(kg·d) groups were significantly increased on endometrium, while these dose does not change ANGPT-1 expression in 50mg/(kg·d) groups.②Compared to secretory phase, the ANGPT-1 proteinum expression in 100, 200mg/(kg·d) groups were much more increased than proliferative phase. Conclusion:①MXC in 100, 200mg/(kg·d) could increase the ANGPT-1 proteinum expression on endometrium;②The increase on proliferative phase of ANGPT-1 proteinum expression in 100, 200mg/(kg·d)groups might display an estrogenic dependence, which might facilitate the recovery of endometrium, or might facilitate the abnomal angiogenesis of endometrium, e.g excess hyperplasia endometrium and endometrial cancer; but the increase on secretory phase might through VSMC proliferation to facilitate maturation stabilization of helicine artery.Part 3. Effects of methoxychlor on endometrium of ANGPT-1 mRNA expression in female mice We erected the model on intraperitoneal injection MXC just like the first experiment. Then mice were euthanized, then collecting their endometrium and extracting RNA,Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of ANGPT-1 mRNA in mice uterus, calculating relative amount of ANGPT-1 mRNA. Results:MXC in 100, 200mg/(kg·d) groups could obviously super-normal endometrium, but there were no obviously differences in MXC in 50mg/(kg·d)groups. Conclusion: MXC in 100, 200mg/(kg·d)groups could increase ANGPT-1 mRNA expression in female mice on endometrium, and then affected the process of physiology and pathology.