CAJ | 학술논문

目的通过对经典Wnt/β-连环蛋白通路的干涉,观察其对小鼠在位内膜间质细胞相关功能的改变及子宫内膜异位症的模型建立的影响.方法小鼠分组注射Wnt/β-连环蛋白通路激活或阻断剂后取其子宫内膜,一部分用于蛋白免疫印迹,免疫组织化学等方法检测在位内膜相关因子及以Boyden侵袭小室、蛋白免疫印迹检测内膜间质细胞的侵袭能力.其余组织建立子宫内膜异位症模型,并以免疫组化、QPCR检测通路相关因子.结果蛋白免疫印迹提示,激活组β-连环蛋白、GSK3β、APC水平显著高于抑制组(P＜0.01)；Boyden侵袭小室提示激活组(113±12)个的穿膜细胞数明显高于对照组(64±13)个,蛋白免疫印迹检测Boyden小室的基质细胞VEGF及MMP-9的表达,经灰度扫描分析显示,激活组较对照组升高(分别升高35.6％,27.4％)和抑制较对照组下降(分别降低12.3％,30.4％)；免疫组化提示抑制组E-钙黏素蛋白呈阳性表达,激活组VEGF蛋白呈强阳性表达；QPCR检测激活组异位内膜Wnt3,Wnt7,GSK3β,Lef,E-钙黏素浓度高于抑制组(均P＜0.05).结论 Wnt/β-连环蛋白信号通路可能通过促进内膜细胞的增殖,加强内膜异位种植、侵袭、转移和血管生成的能力,从而导致子宫内膜异位症的发生.
목적 통과대경전Wnt/β-련배단백통로적간섭,관찰기대소서재위내막간질세포상관공능적개변급자궁내막이위증적모형건립적영향.방법 소서분조주사Wnt/β-련배단백통로격활혹조단제후취기자궁내막,일부분용우단백면역인적,면역조직화학등방법검측재위내막상관인자급이Boyden침습소실、단백면역인적검측내막간질세포적침습능력.기여조직건립자궁내막이위증모형,병이면역조화、QPCR검측통로상관인자.결과 단백면역인적제시,격활조β-련배단백、GSK3β、APC수평현저고우억제조(P＜0.01)；Boyden침습소실제시격활조(113±12)개적천막세포수명현고우대조조(64±13)개,단백면역인적검측Boyden소실적기질세포VEGF급MMP-9적표체,경회도소묘분석현시,격활조교대조조승고(분별승고35.6％,27.4％)화억제교대조조하강(분별강저12.3％,30.4％)；면역조화제시억제조E-개점소단백정양성표체,격활조VEGF단백정강양성표체；QPCR검측격활조이위내막Wnt3,Wnt7,GSK3β,Lef,E-개점소농도고우억제조(균P＜0.05).결론 Wnt/β-련배단백신호통로가능통과촉진내막세포적증식,가강내막이위충식、침습、전이화혈관생성적능력,종이도치자궁내막이위증적발생.
Objective To employ the classical Wnt/β-catenin signaling pathway interference to explore the effects on the functional changes of eutopic endometrium stromal cells and the differences between endometriosis in a murine model.Methods Two out of three mouse groups received an injection of either Wnt/β-catenin signaling pathway activator or blocker.Later the endometrial tissue samples were obtained to develop endometrial stromal cell cultures for the detection of cell invasion ability via Boyden chamber invasion assay and Western blot (WB).Then the methods of WB and Immunohistochemical staining (IHC)were used to examine the factors of eutopic endometrium.And an endometriosis model was established to investigate the factors of signaling pathway via quantitative polymerase chain reaction ( QPCR ) and IHC.Results According to WB test,the level of β-catenin,GSK-3β and APC in the activation group were significantly higher than in the inhibition group( P ＜ 0.01 ).In Boyden chamber invasion assay,the number of cells on membranes in the trial group was significantly higher than the control group [ (113 ± 12) vs (64 ± 13) ].The expressions of VEGF and MMP-9 in the endometrial stromal cells culture from Boyden chamber assay analyzed via WB were ranked from highest to lowest respectively as activation group (vs control group was 35.6％ and 27.4％ higher),control group and inhibition group (vs control group was 12.3％ and 30.4％ lower).Furthermore,the endometrial E-cadherin and VEGF examined via IHC respectively showed a positive expression in inhibitor group and strong positive expression in activation group.QPCR showed the level of Wnt3,Wnt7,GSK3β,Lef and E-cadherin in the activation group was higher than those in the inhibition group ( P ＜ 0.05 ).Conclusion The intervention of WNT signaling pathway in vivo cause the changes of eutopic endometrial invasion and adhesion function,and further affect the development of endometriosis.Wnt/β-catenin signaling pathway may promote the eutopic endometrial cell proliferation and improve the ability of eutopic endometrial implantation, invasion,metastasis and angiogenesis.