DNA甲基转移酶抑制剂对子宫内膜异位症异位灶的影响  

Effects of DNA Methyltransferase Inhibitors on Ectopic Foci in Rats with Endometriosis

作　　者：张婷[1] 夏良斌[1] 高静[2] 曾莹[1] 王丹[1] 付义霞[1] 

机构地区：[1]武汉大学人民医院,湖北武汉430060 [2]武汉市第一医院,湖北武汉430022

出　　处：《实用妇产科杂志》2016年第1期57-62,共6页Journal of Practical Obstetrics and Gynecology

摘　　要：目的:探讨DNA甲基转移酶抑制剂5-氮-2'-脱氧胞苷对子宫内膜异位症大鼠模型异位灶的影响。方法:根据大鼠自体移植方法构建大鼠子宫内膜异位症模型。4周后二次开腹判断造模是否成功,用游标卡尺测量记录异位灶体积后将大鼠随机分为实验组(n=12)、磷酸盐缓冲液(PBS)组(n=12)和对照组(n=11),同时选取未造模的正常大鼠作为空白组(n=10)。实验组大鼠腹腔注射5-氮-2'-脱氧胞苷(2.5 mg/kg·d),PBS组注射等量PBS溶液对照组和空白组未用药。4周后取大鼠腹腔冲洗液及异位灶标本后处死大鼠,再次用游标卡尺测量备组异位灶体积大小。异位灶组织常规HE切片后显微镜下观察组织形态结构。应用免疫组织化学染色方法和Western Blot法检测备组异位灶内DNA甲基转移酶1(DNMT1)的表达情况。收集腹腔冲洗液用Elisa法检测肿瘤坏死因子(TNF-α)含量。结果:①50只大鼠根据大鼠子宫内膜自体移植方法成功35例,造模成功率为70%(35/50)。②实验组大鼠经药物处理后异位灶体积明显缩小(处理前后病灶体积分别为51.08±12.73 mm^3和5.56±3.68 mm^3),差异有统计学意义(P<0.01)。③免疫组化检测实验组、PBS组、对照组异位灶组织中DNMT1表达的组织学评分分别为2.30±0.97分、3.80±1.77分、4.07±1.92分;Western Blot法检测实验组、PBS组、对照组异位灶组织中DNMT1蛋白的相对表达量分别为0.17±0.39、0.29±0.17、0.3 1±0.41。实验组异位灶组织中DNMT1表达低于PBS组及对照组差异有统计学意义(P<0.05)。④实验组大鼠腹腔冲洗液中TNF-α含量较PBS组及对照组降低,实验组、PBS组、对照组均较空白组均升高差异有统计学意义(P<0.05)。结论:DNA甲基转移酶抑制剂能明显减少异位灶中DNMT1的表达及腹腔冲洗液中TNF-α含量,缩小异位灶体积,促进异位灶萎缩,为子宫内膜异位症的靶向治疗提供了依据。Objective：To explore the effects of DNA methyltransferase inhibitors on the ectopic foci in rats with endometriosis.Methods：The rat model of endometriosis by autologous transplantation was established.After four weeks of transplantation,succefful establishment of the model was verified by laparotomy.Following the measurement of the volume by the vernier caliper,the rats were divided into three groups randomly：Experimental Group（treated with 5-aza-2＇-deoxycytidine,2.5 mg/kg·d,n = 12）,PBS Group（treated with equal volum of PBS buffer,n = 12）,Control Group（without treatment,n = 11） and Blank Group（health rats without laparotomy,n = 10）.The volum of ectopic foci were measured after four weeks,followed by collection of the peritoneal washing liquid and ectopic foci.The morphosis of endometriosis tissue was observed by microscopy.The expression of DNA methyltransferase 1 were examined by Western Blotting and immunohistochemistry（IHC）.The expression of TNF-α in peritoneal washing liquid was examined by enzyme-linked immune sorbent assay（Elisa）.Results：①Of all 50 rats,35 cases were succeeded to establish the model by the autologous transplantation method,the achievement ratio being70%（35/50）.②The lesion volume after treatment in experimental group was decreased compared to that before treatment,with a significant difference（51.08±12.73 mm^3 v.s.5.56±3.68 mm^3,P0.01）.③The expression of DNA methyltransferase 1 were statistically significant decreased in experimental group after medication（P0.05）.The IHC histological scores of DNA methyltransferase 1 in ectopic foci tissue was 2.30±0.97（experimental group）,3.80±1.77（PBS group）,and 4.07±1.92（control group）,respectively.The relative expression of DNA methyltransferase 1 of ectopic foci tissue by Western Blot was 0.17±0.39（experimental group）,0.29±0.17（PBS group）,and 0.31±0.41（control group）,respectively.Both differences were statisticlly significant.④After intervention,the TNF-α level of p

关 键 词：子宫内膜异位症 DNA甲基化 大鼠模型 DNA甲基转移酶抑制剂 

分 类 号：R711.71[医药卫生—妇产科学]