CAJ | 학술논문

目的：研究紧密连接蛋白 ZO-1（zonula occludens proteins）、Occludin 在间质性膀胱炎（IC）与正常对照（NC）之间的表达差异并分析其与抗增殖因子（APF）的关联性，探讨它们在IC发病中的作用，为阐明IC的发病机理提供新的思路。方法通过组织块培养法培养正常及IC膀胱上皮细胞，利用流式细胞检测法测定其纯度；运用离子交换层析等技术纯化 APF，检测 APF 对膀胱上皮细胞的抑制效果并评价 APF 的活性；以纯化的 APF 培养 IC 膀胱上皮细胞，以 PBS 处理组为对照，采用Western-blot方法检测ZO-1及Occludin在处理前后的表达变化并分析APF与Occludin，ZO-1之间的关系。结果组织块培养法成功培养出膀胱上皮细胞，纯化的 APF 对膀胱上皮细胞生长的抑制率为78.5%；IC患者的ZO-1、Occludin的表达水平均低于正常组，且在经纯化的APF培养处理的膀胱上皮细胞中，ZO-1、Occludin的表达明显较PBS对照组低，P<0.01）。结论紧密连接蛋白 ZO-1、Occludin 在IC 中呈低表达，纯化的 APF 可进一步降低 ZO-1、Occludin 的表达水平，提示 APF 在 IC 中的病因作用可能是通过抑制紧密连接蛋白表达而产生，即APF可能通过下调紧密连接蛋白ZO-1、Occludin，破坏膀胱黏膜紧密连接结构，提高膀胱黏膜上皮的通透性，从而促进间质性膀胱炎的发生。
목적：연구긴밀련접단백 ZO-1（zonula occludens proteins）、Occludin 재간질성방광염（IC）여정상대조（NC）지간적표체차이병분석기여항증식인자（APF）적관련성，탐토타문재IC발병중적작용，위천명IC적발병궤리제공신적사로。방법통과조직괴배양법배양정상급IC방광상피세포，이용류식세포검측법측정기순도；운용리자교환층석등기술순화 APF，검측 APF 대방광상피세포적억제효과병평개 APF 적활성；이순화적 APF 배양 IC 방광상피세포，이 PBS 처리조위대조，채용Western-blot방법검측ZO-1급Occludin재처리전후적표체변화병분석APF여Occludin，ZO-1지간적관계。결과조직괴배양법성공배양출방광상피세포，순화적 APF 대방광상피세포생장적억제솔위78.5%；IC환자적ZO-1、Occludin적표체수평균저우정상조，차재경순화적APF배양처리적방광상피세포중，ZO-1、Occludin적표체명현교PBS대조조저，P<0.01）。결론긴밀련접단백 ZO-1、Occludin 재IC 중정저표체，순화적 APF 가진일보강저 ZO-1、Occludin 적표체수평，제시 APF 재 IC 중적병인작용가능시통과억제긴밀련접단백표체이산생，즉APF가능통과하조긴밀련접단백ZO-1、Occludin，파배방광점막긴밀련접결구，제고방광점막상피적통투성，종이촉진간질성방광염적발생。
Objective To compare the expression of Occludin, Zo-1 in interstitial cystitis (IC) and normal controls groups, analysing their relationship with antiproliferative factor (APF) in order to explore their role in the pathogenesis of IC. Methods Normal and IC bladder epithelial cells were cultured through bladder tissue and their purity were detemined by flow cytometry method. Immunohistochemistry and Western-blot was used to detect the expression of occludin and ZO-1 in IC group and normal group; After that, the bladder epithelial cells were treated with purified APF, and then we analyse the relationship between of APF and occludin, ZO- 1 compared to the PBS- treated group. Results The expression of occludin, ZO- 1 in IC patients were lower than the normal group. What was more,their expression was significantly lower in purified APF-treated bladder epithelial cells than in the PBS-treated control group. Conclusions The occludin, ZO- 1 was low- expression in the bladder epithelial cells of IC group and purified APF can further reduced their expression in the bladder epithelial cells, indicating that APF may damage bladder mucosa tight junction structure and improve bladder epithelial permeability by inhibiting the expression of claudin and ZO-1, and promote the occurrence of interstitial cystitis.