基于EphA1/EphrinA1反馈环的表达探讨BrMC调控子宫腺肌病的机制  

Based on the expression of EphA1/EphrinA1 feedback loop to explore the regulation mechanism of BrMC on adenomyopathy

作　　者：郑小妹 林叶飞 张韶琼 杨洁 陈曼玲[1] ZHENG Xiao-mei;LIN Ye-fei;ZHANG Shao-qiong;YANG Jie;CHEN Man-ling(Department of Gynecology,the First Affiliated Hospital of Hainan Medical University,Haikou 570100,Hainan,China)

机构地区：[1]海南医学院第一附属医院妇科,海南海口570100

出　　处：《川北医学院学报》2023年第8期1011-1018,共8页Journal of North Sichuan Medical College

基　　金：海南省卫生健康行业科研项目(21A200064)。

摘　　要：目的:探究肝配蛋白A受体1(EphA1)及其配体(EphrinA1)在子宫腺肌病(AM)患者相关组织中的表达情况,并观察8-溴-7-甲氧基白杨素(BrMC)处理对子宫腺肌细胞的影响。方法:采用HE染色对AM患者子宫肌层异位内膜组织、非异位内膜组织及子宫肌瘤患者正常内膜组织的形态学变化进行观察;采用免疫组织化学染色检测3种组织中EphA1与EphrinA1表达情况;分离并培养子宫腺肌细胞,分别使用浓度为2.5、5、10μmol/L BrMC的培养基处理细胞,采用MTT法检测不同处理时间后细胞存活率,处理培养48 h后收集细胞,采用TUNEL染色与Annexin V-FITC/PI法检测细胞凋亡;采用细胞划痕实验检测细胞迁移;采用RT-qPCR和Western blot检测细胞中EphA1、EphrinA1 mRNA与蛋白的表达水平。结果:AM患者异位和非异位子宫内膜组织中EphA1、EphrinA1表达水平均高于对照组(P<0.05);经鉴定发现成功分离子宫腺肌细胞,波形蛋白与α-平滑肌肌动蛋白均表达;经不同浓度BrMC处理后,与对照组比较,各处理组子宫腺肌细胞存活率下降,凋亡率增加,细胞迁移率降低;细胞内EphA1、EphrinA1的mRNA与蛋白相对表达量降低(P<0.05)。结论:EphA1与EphrinA1在子宫腺肌病内膜组织中表达水平升高,经BrMC处理子宫腺肌细胞后,抑制了细胞生长、迁移并促进细胞凋亡,其机制可能与调控EphA1/EphrinA1反馈环相关。Objective:To explore the expression of Ephrin A receptor 1(EphA1)and its ligand(EphrinA1)in related tissues of patients with adenomyosis(adenomyosis,AM),and observe the effect of 8-bromo-7-methoxy(BrMC)treatment on uterine adenomyosis cells.Methods:HE staining was used to observe the morphological changes of ectopic endometrial tissue,non-ectopic endometrial tissue in AM patients and normal endometrial tissue in uterine fibroids patients,and immunohistochemical staining was performed to detect EphA1 and EphrinA1 expressions in the three tissues.Adenomyocytes were isolated and cultured,and treated with medium containing 2.5,5.0 and 10.0μmol/L BrMC,respectively,MTT method was used to detect cell survival rate after different treatment time,after 48 h,TUNEL staining and Annexin V-FITC/PI method were used to detect cell apoptosis,cell scratch test was used to detect the cell migration,real-time fluorescence quantitative PCR and Western blot detection of the expression levels of EphA1,EphrinA1 mRNA and protein in the cells.Results:The expression levels of EphA1 and EphrinA1 in the endometrial tissue of ectopic and non-ectopic AM patients were higher than those in the control group(P<0.05).After identification,it was found that uterine muscle cells were successfully isolated,and both vimentin andαsmooth muscle actin were clearly expressed.After treatment with different concentrations of BrMC,compared with the control group,the survival rate of adenomyosis cells in each treatment group decreased,the apoptosis rate increased,and the cell migration rate decreased,at the same time,intracellular the relative expressions of EphA1 and EphrinA1 mRNA and protein were reduced(P<0.05).Conclusion:The expression of EphA1 and EphrinA1 increased in endometrial tissue of adenomyosis,after BrMC treatment of adenomyosis,it inhibited cell growth,migration and promoted cell apoptosis,the mechanism may be related to the regulation of EphA1/EphrinA1 feedback loop.

关 键 词：子宫腺肌病 8-溴-7-甲氧基白杨素 EphA1 EPHRINA1 

分 类 号：R711.71[医药卫生—妇产科学]