雌激素剥夺状态下表皮生长因子信号系统在内异症发病中的作用  

作　　者：王宇全[1] 尹利荣[1] 郭蕊萌[1] 盛威[1] 

机构地区：[1]天津医科大学第二医院妇科,300211

出　　处：《中华妇产科杂志》2013年第6期447-452,共6页Chinese Journal of Obstetrics and Gynecology

摘　　要：【摘要】目的探讨在雌激素剥夺状态下表皮生长因子（EGF）、表皮生长因子受体（EGFR）、磷酸化细胞外信号调节激酶1／2（p-ERKl／2）在内异症发病中的作用。方法采用细胞培养基快速雌激素剥夺法和对内异症模型裸鼠切除双侧卵巢（去势）的方法，建立体外和体内两种雌激素剥夺的实验研究模型。（1）体外实验：对体外培养的雌激素剥夺状态下的异位子宫内膜细胞按以下不同的处理方法分为4组：①EGF组：加入不同浓度（0．01、0．1、1、10、50、100ng／m1）的EGF培养细胞72h（以浓度为10ng／ml的EGF培养72h的结果代表EGF组的结果）；②EGF＋PD98059组：以5X10。mol／LPD98059[细胞外信号调节激酶（ERK）抑制剂]培养细胞24h后，再以10rig／mlEGF＋5×1012mol／LPD98059培养细胞72h；cr＋ICll82780组：以10μmol／LICll82780（ER抑制剂）培养细胞24h后，再以10ng／mlEGF＋10“mol／LICll82780培养细胞72h；④空白对照组：不进行任何处理。用四甲基偶氮唑盐比色法（MTT）检测处理后各组异位子宫内膜细胞的增殖活性[以吸光度（A）值表示]；蛋白印迹法检测EGF组、EGF＋PD98059组异位子宫内膜细胞中p-ERKl／2的蛋白表达。（2）体内实验：利用随机数字表法将64只内异症模型裸鼠随机分成对照组和去势组，每组各32只，去势组在造模3周后切除双侧卵巢。于造模后的4，6，8，10周，采用蛋白印迹法检测两组裸鼠腹腔病灶中EGF、EGFR、p-ERKl／2的蛋白表达水平。结果（1）细胞增殖活性：EGF组经不同浓度（0．01、0，1、1、10、50、100ng／m1）的EGF作用72h后，细胞增殖活性依次为0．310±0．010、0．340±0．020、0．670±0．010、0．980±0．030、1．360±0．020、1．670±0．020，随EGF浓度的增加，细胞增殖活性逐渐增加。EGF＋PD08059组细胞增殖活性为0．680±0．030，与EGF组（以浓度为10ng／ml的EGF培养72h的结果代表Objective To study the role of epidermal growth factor (EGF),epidermal growth factor receptor(EGFR),extracellular signal-regulated kinase 1/2 (p-ERK1/2) in the pathogenesis of endometriosis under estrogen deprivation conditions.Methods The estrogen was quickly-stripped in medium and the female nude mice were castrated by bilateral oophorectomy to build estrogen deprivation in vitro and in vivo experimental models,respectively.(1) In vitro experiments:according to different treatments the estrogen deprived ectopic endometrial cells were classified into 4 groups:①EGF group:the ectopic endometrial cells were cultured for 72 hours with different concentrations of EGF (0.01,0.1,1,10,50,100 ng/ml),the results of EGF group were represented by the result of cells treated by 10 ng/ml EGF cultured for 72 hours ; ②EGF + PD98059 group:the ectopic endometrial cells were cultured for 72 hours with 5 × 10-2 mol/L PD98059 (inhibitor of ERK),followed by a cultivation for 72 hours treated by 10 ng/ml EGF + 5 × 10-2 mol/L PD98059 ; ③EGF + ICI182780 group:the ectopic endometrial cells were cultured for 72 hours with 10-6 mol/L ICI182780 [inhibitor of estrogen receptor(ER)],followed by a cultivation for 72 hours treated by 10 ng/ml EGF + 10-6 mol/L ICI182780; ④Blank control group:the ectopic endometrial cells were cultured with no treatment.The proliferation activity of ectopic endometrial cells in all groups after treatment were examined by methyl thiazolyl tetrazolium (MTT) method represented by absorbance value (A).The expression of p-ERK1/2 protein were detected by western blot.(2) In vivo experiments:64 female nude mice were randomly divided into control and castration groups (both n =32) using random number chart.The mice in castration group were castrated by bilateral oophorectomy on 3 weeks after the endometriosis model was established.The levels of EGF,EGFR,p-ERK1/2 protein in ectopic lesions of both groups were measured on 4,6,8 and 10 weeks after the endometriosis model was established by western blot.Results (1)

关 键 词：子宫内膜异位症 阉割 雌激素类 表皮生长因子 受体 表皮生长因子 细胞外信号调节MAP激酶类 

分 类 号：R711[医药卫生—妇产科学]