CAJ | 학술논문

目的研究纤溶酶Kringle区缺失突变体(Plm△K)对兔眼玻璃体视网膜界面的作用.方法 Plm△K由毕赤酵母表达的纤溶酶原Kringle区缺失突变体(Plg△K)经组织纤溶酶原激活剂(tPA)激活产生,并通过发色底物S_(2403)测定其蛋白水解活性.0.5、1.0、1.5μmol/min Plm△K 100μL分别注入16只新西兰白兔的玻璃体腔内,注射后1d、7d在光镜下和扫描电镜下检查,并行大体检查、眼部B型超声和光学相干断层扫描(OCT)检查,观察玻璃体视网膜界面情况.结果还原性SDS-PAGE凝胶成像分析证实,Plg△K被激活后产生相对分子质量约26000和5000的2条肽链,且具有纤溶酶活性.4种检测结果均显示Plm△K诱导玻璃体皮质与视网膜的分离.视网膜内表面皮质残留量与Plm△K的剂量呈负相关(r=-0.9516,P=0.048),完全性分离可产生光滑的视网膜内表面.Plm△K注射眼与对照眼的视网膜外层结构均未发现明显的差异.视网膜内表面皮质残留量与药物作用时间有一定的关系,但差异无统计学意义(r=-0.720,P=0.470).对照组和Plm△K处理组间视网膜外层的形态学无明显不同.Plm△K玻璃体注射后未发现视网膜发生药物相关的不良反应.结论玻璃体腔单独注射Plm△K可有效诱导玻璃体与视网膜的完全性分离,而对外层视网膜结构无明显影响.
목적 연구섬용매Kringle구결실돌변체(Plm△K)대토안파리체시망막계면적작용.방법 Plm△K유필적효모표체적섬용매원Kringle구결실돌변체(Plg△K)경조직섬용매원격활제(tPA)격활산생,병통과발색저물S_(2403)측정기단백수해활성.0.5、1.0、1.5μmol/min Plm△K 100μL분별주입16지신서란백토적파리체강내,주사후1d、7d재광경하화소묘전경하검사,병행대체검사、안부B형초성화광학상간단층소묘(OCT)검사,관찰파리체시망막계면정황.결과 환원성SDS-PAGE응효성상분석증실,Plg△K피격활후산생상대분자질량약26000화5000적2조태련,차구유섬용매활성.4충검측결과 균현시Plm△K유도파리체피질여시망막적분리.시망막내표면피질잔류량여Plm△K적제량정부상관(r=-0.9516,P=0.048),완전성분리가산생광활적시망막내표면.Plm△K주사안여대조안적시망막외층결구균미발현명현적차이.시망막내표면피질잔류량여약물작용시간유일정적관계,단차이무통계학의의(r=-0.720,P=0.470).대조조화Plm△K처리조간시망막외층적형태학무명현불동.Plm△K파리체주사후미발현시망막발생약물상관적불량반응.결론 파리체강단독주사Plm△K가유효유도파리체여시망막적완전성분리,이대외층시망막결구무명현영향.
Background The vitreoretinal traction plays a critical role in the formation of macular hole and cystoid macular edema.Enzymatic vitreolysis has potential in relieving vitreoretinal traction as a simple and less invasive method in comparison with pars plane vitrectomy.ObjectiveThis study is to investigate the effects of plasmin mutant with kringle domains deficiency(Plm△K)on vitreoretinal interface in new Zealand white rabbits.Methods Plm△K was prepared through activating plasminogen mutant with Kringle domains deficiency (Plg△K) by tissue plasminogen activator (tPA).100μL of Plm△K at the dose of 0.5,1.0 and 1.5μmol/min was injected respectively into the vitreous of 48 New Zealand white rabbits and 16 eyes for each dose.B-scan and optical coherence tomography (OCT) were performed to detect the structure variety at the vitreoretinal interface in 1 day and 7 days after injection.The gross anatomy analysis with triamcinolone acetonide fine particle suspension,as well as histopathological examinations by scanning electron microscopy,was performed in the different time points mentioned above.Results Two peptide chains were determined with the relative molecular weight about 26000 and 5000 by the gel imaging analysis of reduced SDS-PAGE.Separation of the posterior vitreous cortex from retina was found after intravitreous injection under the B-scan and OCT.The ultrastructure change of vitreoretinal interface as well as the examination of fine particle suspension by triamcinolone acetonide demonstrated the same outcome.The degree of remnants of vitreous cortex showed the negative correlation with the dosage of Plm△K (r=-0.9516,P=0.048).No significant correlation was found between the degree of remnants of vitreous cortex and the action time(r=-0.720,P=0.470).There was no obvious morphological difference in outer layer of retina between control eyes and Plm△K-treated eyes.No drug-related adverse event was found after intravitreous injection of Plm△K.Conclusion Intravitreous injection of Plm△K alone can induce complete separation of vitreous from retina.This procedure is safe and effective.