腹腔注射甲氨蝶呤对小鼠异位内膜生长的影响

目的观察腹腔注射甲氨蝶呤对子宫内膜异位症模型小鼠异位内膜生长的影响,探讨甲氨蝶呤作为难治性子宫内膜异位症治疗药物的可能性。方法2005年6月至2006年5月在复旦大学附属妇产科医院采用增强型绿色荧光蛋白（EGFP）转基因和野生型C57BL/6J小鼠建立子宫内膜异位症模型,2周后对照组（D组）给予等量的生理盐水腹腔注射,实验组分别给予25mg/kg（A组）,50mg/kg（B组）,75mg/kg（C组）的甲氨蝶呤腹腔注射,1周后处死动物,荧光活体成像后比较各组小鼠荧光表达的强弱,取异位病灶、子宫及卵巢行病理检测,流式细胞术检测异位病灶的凋亡情况。结果镜下可见模型小鼠肠管间、腹壁下、肝小叶下或脾周形成绿色团块状或星点状内膜异位病灶,实验组异位灶体积及荧光表达强度明显低于对照组（P〈0.01）,而各实验组异位内膜细胞凋亡率明显高于对照组（P〈0.05）,且这种差别呈剂量依赖（P〈0.05）。结论甲氨蝶呤可抑制子宫内膜异位症模型小鼠异位内膜细胞的生长并诱导其凋亡,且这种作用呈剂量依赖,提示肿瘤化疗药甲氨蝶呤有望用于某些难治性子宫内膜异位症的临床治疗。

Effect of methotrexate injected intraperitoneally on ectopic endometrium of mice model of endometriosis

Objective To observe the effect of methotrexate which injected intraperitoneally on ectopic endometrium of mice model of endometriosis and to investigate the possibility of chemotherapy drug methotrexate which used as clinical remedy for some innocence disease to be used as remedy for refractory endometriosis.Methods From June 2005 to May 2006 in the Obstetrical and Gynecological Hospital of Fudan University.Mice models of endometriosis were established by enhanced green fluorescent protein(EGFP)transgenic and wide-type C57BL/6J mice and two weeks later sodium chloride was injected intraperitoneally into the control group(D group)while methotrexate of 25mg/kg(A group),50mg/kg(B group),75mg/kg(C group)was injected intraperitoneally into the experimental mice,respectively.One week later all model mice were sacrificed and optical density of fluorescence of ectopic endometrium was calculated after t-hat.The ectopic endometrium,uterus and ovaries were examined histologically and FCM analysis was performed to access cell apoptosis.Results Under fluorescent microscope the green cake-like or punctiform ectopic endometrium were found between the intestines,under abdominal wall or under hepatic lobules or spleen in the model mice.The volume and optical density of fluorescence of ectopic endometrium of the control group were significantly higher than that of experimental group(P< 0.01);the cell apoptosis of experimental group was significantly higher than that of control group(P< 0.05)and the difference was dose-dependent(P< 0.05).Conclusion Methotrexate can exert growth-inhibitory and apoptosis-inducing effect on ectopic endometrium of model mice and this effect is dose-dependent.This tells us that chemotherapy drug methotrexate may be used as remedy for some refractory endometriosis.