NF-κBp65蛋白及ENA-78在子宫内膜异位症发病中的作用研究

目的探讨核因子κBp65(NF-κBp65)蛋白及中性粒细胞活化肽-78(ENA-78)在子宫内膜异位症(内异症)发病中的作用。方法选取2008年10月至2009年2月在山西医科大学第二医院手术切除的卵巢子宫内膜异位囊肿25例,留取异位内膜与在位内膜组织,分为异位内膜组和在位内膜组,选取同期正常的子宫内膜22例为对照组。不同浓度的白细胞介素1β(IL-1β)及IL-1β+二硫氨基甲酸肽吡咯烷(PDTC)分别干预体外传代培养至第3代的在位、异位及正常内膜基质细胞。免疫细胞化学方法及酶联免疫吸附实验(ELISA)检测3组子宫内膜基质细胞NF-κBp65蛋白活化程度及其培养上清液中ENA-78浓度。用IL-1β、IL-1β+PDTC检验各组内膜基质细胞分泌ENA-78及NF-κBp65蛋白的活化程度。结果在位及异位子宫内膜基质细胞培养上清液中ENA-78浓度明显高于正常子宫内膜基质细胞,差异有统计学意义(P0.01);在位及异位子宫内膜基质细胞之间比较,异位子宫内膜基质细胞培养上清液中的ENA-78浓度高于在位子宫内膜基质细胞,差异有统计学意义(P0.01),分别给予IL-1β、IL-1β+PDTC干预后,正常子宫内膜基质细胞上清液中的ENA-78浓度无明显变化(P0.05);而在位及异位内膜基质细胞的上清液中,ENA-78浓度在IL-1β干预后最高,明显高于未干预时及IL-1β+PDTC干预后,差异有统计学意义(P0.01)。3组中,在位及异位内膜基质细胞的NF-κBp65蛋白的活化程度显著高于对照组,差异有统计学意义(P0.05);在位及异位子宫内膜基质细胞NF-κBp65蛋白的活化程度IL-1β干预后最高,显著高于未干预时及IL-1β+PDTC干预后,差异有统计学意义(P0.05)。结论 IL-1β可诱导在位及异位子宫内膜基质细胞NF-κBp65活化,使ENA-78的表达增高。PDTC可抑制此作用,说明NF-κBp65是关键性的调控因子,可能在内异症的发病过程中起到"闸门"的作用,对ENA-78以及其他细胞因子起到重要的调控作用。

Study of the effect of NF-κBp65 protein and ENA-78 in the onset of endometriosis.

Abstract： Objective To detect the effect of κBp65 and ENA-78 in the onset of endometriosis. Methods From October 2008 to February 2009 in the Second Hospital of Shanxi Medical University,25 cases undergoing operation resection of ovarian endometriosis cyst were chosen,ectopic and eutopic endometrium tissue was taken,and they were divided into ectopic and eutopic group;select the same normal endometrium of 22 patients as the control group. The eutopic endometrium, ectopic endometrium and thenormal endometrium cells, which were in third passage，were interfered by IL-1β and IL-1β adding PDTC with different concentration. Use ELISA to detect the concentrations of ENA-78 of those three groups，explore the effect of IL-1β and IL-1β adding PDTC on the concentration of ENA-78 of each group.Use immunohistochemistry to detect the activating degree of NF-κBp65 of the endometrial cells in the three groups，and to detect the change of NF-κBp65’s activating degrees after intervention of IL-1β and IL-1β adding PDTC. Results The eutopic endometrium and the ectopic endometrium concentration of the ENA-78 in the cells culture media supernatant was remarkably higher than the normal endometrium.There was a remarkable difference（P<0.01）.The ectopic endometrium concentration of the ENA-78 in the cells culture media supernatant was remarkably higher than the eutopic endometrium.There was a remarkable difference（P<0.01）.After the intervention，the concentration of the ENA-78 in the normal endometrial cells culture media supernatant had no remarkable change.There was no remarkable difference（P>0.05）.In the eutopic endometrium and the ectopic endometrium cells culture media supernatant，the concentration of the ENA-78 in the group intervened with IL-1β was the highest.It was remarkably higher than the control group and the group intervened by IL-1β+PDTC.There was a remarkable difference（P<0.01）. In three groups，the NF-κBp65’s activating degrees of eutopic and the ectopic endometrium were remarkably higher than the normal endometrium.The difference was statistically significant （P<0.05）；for the NF-κBp65’s activating degrees in the eutopic endometrium and the ectopic endometrium cells，the IL-1β group was the highest，which was remarkably higher than the controlled group and the group intervened by IL-1β+PDTC. The difference was statistically significant）（P<0.05）. Conclusion IL-1β can induce the activation of NF-κBp65 on eutopic and ectopic endometrium cells.It can strengthen the expression of ENA-78，while PDTC inhibits this effect.It suggests that as a key controlling agent，NF-κBp65 may play a “penstock” role in the onset of EMs，and it may have a controlling effect on ENA-78 and other cytokines.