复宫宁对大鼠异位子宫内膜细胞形态及caspase-3和Bcl-2蛋白表达的影响

Effect of Fugongning on morphology and protein expression of caspase-3 and Bcl-2 in rat endometriotic cells

目的:研究中药复宫宁制剂对子宫内膜异位症(EMT)大鼠异位子宫内膜细胞凋亡、形态学改变及caspase-3和Bcl-2蛋白表达的影响,探讨复宫宁治疗EMT的作用机制。方法:用自体子宫内膜移植法复制EMT大鼠模型,将EMT大鼠随机分为模型组、达那唑治疗组、复宫宁治疗1组和复宫宁治疗2组,灌胃给药4周,观察各给药组及模型组异位病灶体积大小,采用HE染色和透射电镜技术观察各给药组及模型组异位内膜细胞形态结构的变化。应用免疫组化方法观察各组caspase-3和Bcl-2蛋白的表达情况。结果:复宫宁治疗组中,异位内膜体积显著缩小(P<0.01),病灶生长明显受抑制。异位内膜腺上皮层明显变薄,腺体减少;细胞呈矮柱状甚至扁平状排列松散,并见较多凋亡现象和间质纤维化,而在位内膜未见明显改变。超微结构可见复宫宁组与达那唑组胞体收缩,失去微绒毛,胞浆浓缩,核染色质密度增高并凝聚于核膜周边,核仁裂解;细胞膜内陷和凋亡小体形成;线粒体肿胀,线粒体嵴变低及空泡变性。免疫组化结果显示,模型组异位内膜Bcl-2蛋白表达增强;复宫宁组异位内膜细胞活化caspase-3蛋白表达增高(P<0.01),而Bcl-2蛋白表达降低(P<0.05)。结论:复宫宁对大鼠EMT有明显的抑制作用,其作用机制与其诱导细胞凋亡有关。复宫宁通过激活细胞中的caspase及降低Bcl-2蛋白在EMT中的高表达,诱导异位内膜细胞的凋亡,可能是其重要机制之一。

AIM： To investigate the effect of Fugongning （FGN） on treating endometriosis （EMT） by observ- ing the changes of histopathological morphology and ultrastructure, and the expression of caspase-3 and Bcl-2 in an endom- etriotic rat model. METHODS： The rat model of EMT was established by autotransplantation of endometrium, and the model rats were randomly divided into 4 groups： EMT group, danazol group, FGN 1 group and FGN 2 group. The rats were treated with normal saline in EMT group, danazol in danazol group and different doses of FGN in FGN 1 group and FGN 2 group by intragastric administration for 4 weeks. After 4 weeks of treatment, the size of the lesion and histopathological morphology of ectopic endometrial tissues were observed under transmission electron microscope. Using immunohistochemis- try method, the protein expression of caspase-3 and Bcl-2 in endometrium and endometriotic tissues was determined. RE- SULTS： The size of transplanted object in FGN treatment groups was much smaller than that in control group, and the growth of focus was inhibited. Under the electron microscope, the ectopic endometrial tissues in FGN treatment groups and damazol group shrinked, glandular epithelium became thinner, and the number of the glands decreased. Ectopic endome- trim cells were shot-cylindrical even tabular with incompact arrangement, and apoptosis and interstitial fibrosis were observed. Eutopic endometria didn＇t show any pathological changes. Besides, mitoehondrion swelling and low,disappeared or vacuolated mitochondrial crista in ectopie endometrial cells were observed. Immunohistochemistry results showed that higher expression of Bcl-2 protein in eetopic endometrial tissue in EMT group was observed. The protein level of caspase-3 in the ec- topic endometrial cells in FGN treatment groups increased, while the protein expression of Bel-2 decreased, thus promoting the apoptosis of the ectopic endometrial ceils. CONCLUSION： FGN inhibits the proliferation of ectopie endometrium by inducing the apoptosis of endometriotic cells through activating caspase activity and reducing Bel-2 expression in the cells.