基于调控HIF-1α表达探讨温经汤改善大鼠子宫内膜异位症的机制  

Mechanism of Wenjingtang on Improving Endometriosis in Rats Based on Regulating HIF-1α Expression

作　　者：任艳青[1] 成秀梅[1] 方惠敏 王迪 李新华[1] REN Yan-qing;CHENG Xiu-mei;FANG Hui-min;WANG Di;LI Xin-hua(Hebei University of Chinese Medicine,Shijiazhuang 050200,China)

机构地区：[1]河北中医学院,石家庄050200

出　　处：《中国实验方剂学杂志》2020年第23期63-70,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82004400);河北省自然科学基金项目(H2020423069);河北中医学院科研能力提升项目(KTZ2019003,KTY2019065);河北中医学院优秀青年教师基础研究计划项目(YQ2019009)。

摘　　要：目的:观察温经汤对子宫内膜异位症(EMs)大鼠缺氧诱导因子-1α(HIF-1α)表达及线粒体超微结构的影响,探讨温经汤治疗EMs的作用机制。方法:采用自体内膜移植法复制大鼠EMs模型,小动物超声成像系统检测异位病灶体积,依据体积均衡原则随机分为模型组、温经汤低、中、高剂量组(4.85,9.7,19.4 g·kg-1)及孕三烯酮组(0.25 mg·kg-1),每组10只,另设假手术组10只。药物治疗6周后,超声成像系统及卡尺分别测量异位病灶体积,苏木素-伊红(HE)染色观察异位内膜组织形态,酶联免疫吸附测定(ELISA)检测腹腔液白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α),转化生长因子-β1(TGF-β1)水平,实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测在位或异位内膜组织HIF-1αmRNA及蛋白表达,透射电镜观察异位内膜线粒体超微结构。结果:与假手术组比较,模型组可见异位病灶生成,呈囊泡样结构,病灶内可见典型的子宫内膜组织形态,IL-1β,TNF-α,TGF-β1含量显著升高(P<0.01),HIF-1αmRNA及蛋白表达水平明显升高(P<0.05,P<0.01),透射电镜可见异位内膜胞浆中线粒体数量增加,结构完整;与模型组比较,温经汤各剂量组异位病灶体积显著降低(P<0.01),且超声检测结果与卡尺测量结果大体一致,HE染色可见异位内膜柱状上皮细胞破损或脱落、间质细胞疏松;温经汤各剂量组TNF-α含量明显降低,温经汤中、高剂量组IL-1β,TGF-β1含量明显降低(P<0.05,P<0.01);温经汤各剂量组还可明显降低HIF-1αmRNA及蛋白表达(P<0.05,P<0.01);且温经汤治疗后异位内膜线粒体明显肿胀,嵴断裂甚至消失,部分线粒体空泡变性,外膜破裂。结论:温经汤对大鼠实验性EMs具有较好的治疗作用,机制与降低HIF-1α表达,改善异位病灶乏氧,诱导异位内膜线粒体损伤有关。Objective: To observe the effect of Wenjingtang on the expression of hypoxia inducible factor-1α(HIF-1α)and ultrastructure of mitochondria in rats with endometriosis(EMs),and to explore the mechanism of Wenjingtang in the treatment of EMs. Method: The EMs model rat was established by autogenous intimal transplantation,and the volume of ectopic lesions was measured by small animal ultrasound imaging system. According to the volume of ectopic lesions,rats successfully modeled were randomly divided into model group,low,middle ang high dose Wenjingtang group(4.85,9.7,19.4 g·kg-1)and gestrinone group(0.25 mg·kg-1),10 in each group,another 10 rats were recruited in a sham operation group. After 6 weeks of drug treatment,the volume of ectopic lesions was measured by ultrasound imaging system and caliper,the morphology of ectopic endometrium was observed by hematoxylin-eosin(HE)staining,the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)in peritoneal fluid were measured by enzyme linked immunosorbent assay(ELISA),the mRNA and protein expression of HIF-1αin eutopic or ectopic endometrial tissues were detected by Real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)and Western blot,and the ultrastructure of mitochondria in ectopic endometrium was observed by transmission electron microscope(TEM). Result: Compared with sham operation group,ectopic lesions were found in model group,showing vesicular structure,and the typical endometrial tissue morphology was found in the lesions. The contents of IL-1β,TNF-α,TGF-β1 and the expression of HIF-1α mRNA and protein were significantly increased than those in the sham operation group(P<0.05,P<0.01). TEM showed that the number of mitochondria in the cytoplasm of ectopic endometrium increased and the structure was intact.Compared with model group,the volume of ectopic focus was significantly decreased in all Wenjingtang groups(P<0.01),and the results of ultrasonic examination were basically co

关 键 词：温经汤 子宫内膜异位症 缺氧诱导因子-1Α 异位病灶体积 小动物超声成像 线粒体超微结构 

分 类 号：R2-0[医药卫生—中医学] R289