The Study Effect Of Methoxychlor On The Biological Characteristics Of Human Cytotrophoblast

The methoxychlor is one of enviroment endocrine disrupting chemicals and chemical name is 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane,it has been widely used as a replacement for DDT due to its lower toxicityt to mammals and its shorter half-life. Methoxychlor was detected in soil,river,animals and plants even in pregnant women,placenta,cord blood and newborn infant. Methoxychlor can disrupt function of male testiculus and epididymis, reduce spermatozoa; methoxychlor can also make ovarian follicle atresia,uterus protein expression changed. Cytotrophoblast include extravillous trophoblast and villous trophoblast ,their biological function are important in pregnancy and its maintenance. We firstly primary culture cytotrophoblast and characterization, then invested their biological characteristics using transwell , RT-PCR,Wsetern-blot and radio-immunity.AIM: 1 To observe the effect of Methoxychlor on the invasiveness of extravillous trophoblast in vitro.2 To observe the effect of Methoxychlor on the immune tolerance of extravillous trophoblast in vitro. 3 To observe the effect of Methoxychlor on human chorionic gonadotropin and progesterone secreted by villous trophoblast.METHOD: 1 Obtained cytotrophoblast subpopulation cells from first-trimester chorionic villi (5-10) weeks by sequential triypsinization and purified ,isolated by flow cytometry. All trophoblast subpopulations was identitied by lightmicroscope, immunocytochemistry, transmission electron microscope. 2 Extravillous trophoblast incubation with 1μmol/L methoxychlor after 24 hours. The effect of Methoxychlor on the invasiveness of extravillous trophoblast was investigated with transwell, .3 The effect of methoxychlor on the expression of MMP-2 and MMP-9 in extravillous trophoblast was investigated with western blot. 4. The effect of methoxychlor on the change of HLA-G mRNA and protein expression in extravillous trophoblast was investigated with reverse transcription-PCR and western blot. 5 Using radio-immunity detected human chorionic gonadotropin and progesterone secreted by villous trophoblst after cultured with 1μmol/L methoxychlor 48h.RESULT: 1 Cells obtained from trypsinization and flow cytometer were extravillous trophoblast and villous trophoblast after characterization. 2 methoxychlor can inhibite the invasiveness of the extravillous trophoblast, 3 The extravillous trophoblast incubation with 1μmol/L methoxychlor significant drease expression of MMP-2 and MMP-9 in western blot experiments. 4 The extravillous trophoblast incubation with 1μmol/L methoxychlor significant drease expression of HLA-G mRNA and protein expression. 5 Methoxychlor can decrease villous trophoblast secreted human chorionic gonadotropin and progesterone.CONCLUSION: 1. trypsinization and flow cytometer can obtain extravillous trophoblast and villous trophoblast. 2 Methoxychlor can inhibite the invasiveness of the extravillous trophoblast, this anti-invasive activity may be the results of decrease in MMP-2 and MMP-9 expression in extravillous trophoblast. 3 Methoxychlor can decrease HLA-G protein and mRNA expression in extravillous trophoblast. 4 Methoxychlor can decrease villous trophoblast secreted human chorionic gonadotropin and progesterone.