CAJ | 학술논문

目的：本研究将探讨在再灌注过程中，MMP‐2和 MMP‐9的激活是否参与claudin‐5和occludin蛋白的降解。方法：选取再灌注3h这个再灌注时间点进行研究。通过明胶酶谱法，确定MMPs的活性来源；通过Western blot测定claudin‐5，occludin在缺血侧、非缺血侧蛋白水平；通过使用MMPs抑制剂GM6001，进一步验证明胶酶（MMP‐2和MMP‐9）在脑缺血再灌注过程中对claudin‐5和occludin蛋白降解的影响。结果：再灌注3h ，明胶酶谱法显示MMP‐2比MMP‐9有较大的增幅。可见，在再灌注3h ，MMP‐2是最主要的酶源。在缺血侧，可见claudin‐5和occludin蛋白降解片段。MMPs抑制剂GM6001使用可以完全抑制缺血侧及非缺血侧MMP‐2和MMP‐9的活性，并抑制claudin‐5和occludin蛋白的降解。结论：MMP‐2和MMP‐9介导的claudin‐5和occludin蛋白降解是与再灌注损伤相关的血脑屏障破坏的一个重要机制。
목적：본연구장탐토재재관주과정중，MMP‐2화 MMP‐9적격활시부삼여claudin‐5화occludin단백적강해。방법：선취재관주3h저개재관주시간점진행연구。통과명효매보법，학정MMPs적활성래원；통과Western blot측정claudin‐5，occludin재결혈측、비결혈측단백수평；통과사용MMPs억제제GM6001，진일보험증명효매（MMP‐2화MMP‐9）재뇌결혈재관주과정중대claudin‐5화occludin단백강해적영향。결과：재관주3h ，명효매보법현시MMP‐2비MMP‐9유교대적증폭。가견，재재관주3h ，MMP‐2시최주요적매원。재결혈측，가견claudin‐5화occludin단백강해편단。MMPs억제제GM6001사용가이완전억제결혈측급비결혈측MMP‐2화MMP‐9적활성，병억제claudin‐5화occludin단백적강해。결론：MMP‐2화MMP‐9개도적claudin‐5화occludin단백강해시여재관주손상상관적혈뇌병장파배적일개중요궤제。
Objective :In this study ,we are interested in whether the activated MMP‐2 and MMP‐9 are involved in the degradation of claudin‐5 and occludin proteins .Methods :First ,by gelatin zymography ,we quantified the activities of MMP‐2 and MMP‐9 in the brain of rats subjected to 2h of middle cerebral artery occlusion followed by 3h of reperfu‐sion .Then ,we future investigate the role of MMP‐2 and MMP‐9 activation in the degradation process of claudin‐5 and occludin proteins .Finally ,using MMPs inhibitor GM6001 ,we further study and verify the effect of gelatinases on the degradation of claudin‐5 and occludin proteins after 3h of reperfusion .Results:Gelatin zymography showed greater in‐creases in MMP‐2 than in MMP‐9 .MMP‐2 was the main enzymatic source at 3h reperfusion .claudin‐5 and occludin ex‐pression decreased at 3h in both hemispheres ,with fragments of both proteins seen in the blots at 3h on the ischemic side .MMPs inhibitor GM6001 completely blocked gelatin cleavage by the improved activities of MMP‐2 and MMP‐9 , and prevent the degradation of claudin‐5 and occludin proteins during the early stage of cerebral ischemia reperfusion , respectively .Conclusion:Our study suggests that MMP‐2 and MMP‐9 mediated degradation of the claudin‐5 and occlu‐din proteins may represent an important mechanism for reperfusion‐associated BBB disruption .