ERK_(1/2)和NF-κBp65在EMs模型大鼠内膜中的表达及意义

The expression and significance of extracellular regulatory protein kinase and nuclear factorκB in the endometrium tissue of rats with endometriosis

目的:探讨细胞外调节蛋白激酶(ERK_(1/2))和核因子κB(NF-κBp65)在子宫内膜异位症(EMs)模型大鼠内膜组织中的表达及其与增殖和侵袭的关系。方法:选取雌性未孕SD大鼠20只,自体移植法诱导建立EMs大鼠模型,其中对照组10只。采用免疫组织化学法(IHC)及Western blot法检测各组大鼠子宫内膜组织中ERK_(1/2)和NF-κBp65蛋白表达;增殖相关蛋白(PCNA)、侵袭相关蛋白(MMP9)表达,验证异位病灶组织增殖和侵袭性的变化。结果:EMs模型大鼠病灶局部可见移植物体积增大呈透明或者半透明,内含淡黄色清亮或者咖啡色液体的囊状结构,表面及周围可见血管形成。苏木素伊红染色可见异位病灶组织内腺体、间质或者腺上皮细胞生长。免疫组化结果显示ERK_(1/2)主要表达于子宫内膜腺上皮细胞的胞浆中,间质内少量表达,胞核内极少量表达。NF-κBp65主要表达于子宫内膜腺体和间质的胞核,胞浆中少量表达。Western blot结果显示,EMs模型大鼠异位组及在位内膜组ERK_(1/2)和NF-κBp65蛋白表达水平高于对照组,且EMs异位内膜组表达水平高于EMs在位内膜组(P<0.05);与对照组相比,PCNA和MMP9蛋白在EMs模型大鼠异位组及在位内膜组的表达明显增高,且EMs异位内膜组表达高于EMs在位内膜组(P<0.05)。Pearson相关分析显示ERK_(1/2)和NF-κBp65蛋白表达水平呈正相关(异位内膜组r=0.657,P<0.05;在位内膜组r=0.709,P<0.05)。结论:ERK_(1/2)与NF-κBp65在EMs大鼠模型在位内膜及异位内膜组织中表达增高,可能影响异位内膜的增殖、侵袭能力,继而促进了EMs的发生发展。

Objective:To investigate the expressions of extracellular regulatory protein kinase(ERK_(1/2))and nuclear factorκB(NF-κBp65)in(EMs)model rat endometrial tissue,and to study their association with the proliferation and invasion of endometriosis cell.Methods:20 non-pregnant female SD rats were selected,and among them,the autologous transplantation of endometrium was used to induce the establishment of 10 models of rats with Ems and another 10 rats were as control.The expression of ERK_(1/2) and NF-κBp65 protein in rat endometrial tissue was detected by immunohistochemistry(IHC)and Western blot.The expressions of proliferating cell nuclear antigen(PCNA)and matrix metalloproteinase 9(MMP9)were used to verify the changes of proliferation and invasiveness of lesions tissue of the ectopic endometrium.Results:In the lesions of model rats with EMs,the enlarged implantation was seen to be transparent or translucent cystic structure,containing light yellow clear or coffee-colored liquid,and blood vessel formation was observed on the surface of cystic structure and around.After hematoxylin eosin stain,the growth of glandular,stroma,or glandular epithelial cells were observed in the ectopic lesion.Immunohistochemical results showed that ERK_(1/2) was mainly expressed in the cytoplasm of endometrial glandular epithelial cells,and was less expression in the stroma of the cells,and had a very small amount in the nucleus of the cells.NF-κBp65 was mainly expressed in the nuclei of endometrial glands cells and stroma of the cells,and was less expression in the cytoplasm of the cells.Western blot results showed that the expression levels of ERK_(1/2) and NF-κBp65 protein in ectopic endometrium and eutopic endometrium of the model rats with Ems were significantly higher than those in normal endometrium of the rats,and which in ectopic endometrium of the model rats with Ems were significantly higher than those in eutopic endometrium of the model rats with Ems(P<0.05).The expression levels of PCNA and MMP9 protein in ectopic endometrium and normal endometrium of the model rats of Ems were significantly higher than those in normal endometrium of the rats,and which in ectopic endometrium of the model rats with Ems were significantly higher than those in eutopic endometrium of the model rats with Ems(P<0.05).Pearson correlation analysis showed that there was positive correlation between ERK_(1/2) level and NF-κBp65 protein expression level(r=0.657,P<0.05)and in eutopic endometrium of the model rats of Ems(r=0.709,P<0.05).Conclusion:The expressions of ERK_(1/2) and NF-κBp65 in the eutopic and ectopic endometrial tissues of the EMs rat model increase,which may improve the occurrence and development of EMs by affecting the proliferation and invasion ability of the ectopic endometrium.