The Effect Of P-glycoprotein, EMMPRIN And MMPs In Villi On Abnormal Uterine Bleeding After Medical Abortion With Mifepristone And Misoprostol

Part I The relationship of P-glycoprotein in villi and abnormal uterine bleeding after medical abortion with mifepristone and misoprostolObjectives:To investigate the relationship of P-glycoprotein and abnormal uterine bleeding after medical abortion by analyzed the expression level of P-glycoprotein in the villi, and the concentration of mifepristone in trophoblast cells of early pregnancy in vitro.Materials and Methods:Women who voluntarily wanted to terminate early pregnancy by medical abortion with mifepristone and misoprostol were recruited. The villi discharged naturally on the day using misoprostol were collected. According to the follow-up, women were divided into two groups which were completed abortion group (37 cases) and bleeding group (37 cases). The location of P-glycoprotein in the villi were examined by immunohistochemistry. The expression levels of P-glycoprotein in the villi of two groups were tested by Western Blotting. In vitro, trophoblasts were cultured with CsA (20μM and 40μM) and mifepristone (0.5μM,1.25μM and 2.5μM). The concentration of mifeprsitone in trophoblasts was detected by HPLC.Results:P-glycoprotein was located in the membrance of syncytiotrophoblast. The expression level of P-glycoprotein in bleeding group was higher than that in completed abortion group(p<0.05). There were no significant differences among three groups (20μM CsA group,40μM CsA group and Control group) in every concentration of mifepristone (0.5μM,1.25μM and 2.5μM) (p>0.05).Conclusions:The increased expression level of P-glycoprotein in the villi may be related to abnormal uterine bleeding after medical abortion. In vitro, P-glycoprotein did not pump the mifepristone out of the trophoblast to reduce the concentration of mifepristone in trophoblasts, namely mifepristone may be not the substrate of P-glycoprotein.Part II The effect of MMP-2, MMP-9 and EMMPRIN in villi on abnormal uterine bleeding after medical abortion with mifepristone and misoprostolObjectives:To investigate the effect of MMP-2, MMP-9 and EMMPRIN on abnormal uterine bleeding after medical abortion with mifepristone and misoprostolMaterials and Methods:Women who voluntarily wanted to terminate early pregnancy by medical abortion with mifepristone and misoprostol were recruited. The villi discharged naturally on the day using misoprostol were collected. According to the follow-up, women were divided into two groups which were completed abortion group (37 cases) and bleeding group (37 cases). The expression levels of MMP-2 and MMP-9 proteins were examined by immunohistochemistry. The activities of MMP-2 and MMP-9 were detected by gelatin zymography. The location of EMMPRIN in the villi were examined by immunohistochemistry. The expression levels of EMMPRIN in the villi of two groups were tested by Western Blotting.Results:There was no significant difference of MMP-2 protein expression levels in two groups (p>0.05); the expression level of MMP-9 protein in bleeding group was higher than that in completed abortion group (p<0.001). The activities of MMP-2 and MMP-9 in two groups were not detected. The expression of EMMPRIN was located in the membrance of syncytiotrophoblast and cytotrophoblast. The expression level of EMMPRIN in bleeding group was lower than that in completed abortion group (p＜0.05).Conclusions:The decreased expression level of EMMPRIN protein and the increased expression level of MMP-9 in the villi may be related to abnormal uterine bleeding after medical abortion; but the expression level of MMP-2 in the villi may be not related to abnormal uterine bleeding after medical abortion.PartⅢThe interaction of P-glycoprotein and EMMPRIN in trophoblasts of early pregnancy in vitroObjectives:To investigate the interaction of P-glycoprotein and EMMPRIN in trophoblasts of early pregnancy in vitro.Materials and Methods Women who voluntarily wanted to terminate early pregnancy by surgical abortion were recruited. The villi were collected and cultured in trophoblast explant. Under the presence of CsA (20μM group and 40μM group), trophoblast explant was cultured for 24 hours and 48 hours, the expression level of EMMPRIN protein was analyzed by Western Blotting. Under the presence of MEM-M6/1 (2.5μg/ml group and 5.0μg/ml group), trophoblast explant was cultured for 24 hours and 48 hours, the expression level of P-glycoprotein protein was analyzed by Western Blotting.Results:Under the presence of CsA (20μM CsA group and 40μM CsA group), cultured for 24 hours or 48 hours, there were no significant differences of the expression levels of EMMPRIN protein among 20μM CsA group,40μM CsA group and control group(p>0.05). Under the presence of MEM-M6/1 (2.5μg/ml MEM-M6/1 group and 5.0μg/ml MEM-M6/1 group), the expression levels of P-glycoprotein protein in two concentrations of MEM-M6/1 groups were lower than that in control group at 24 hours' culture. There were significant differences among three groups(2.5μg/ml MEM-M6/1 group,5.0μg/ml MEM-M6/1 group and control group) (p<0.05); there were no significant differences among three groups at 48 hours'culture(p>0.05).Conclusions:In vitro, EMMPRIN protein maybe regulate the expression level of P-glycoprotein protein, but P-glycoprotein protein maybe not regulate the expression level of EMMPRIN protein.