全视网膜激光光凝术后玻璃体内过氧化物的荧光光度检测

Fluorophotometric detection of intravitreal peroxides after panretinal laser photocoagulation.

作者信息

Taguchi H, Ogura Y, Takanashi T, Hashizoe M, Honda Y

机构信息

Department of Ophthalmology and Visual Sciences, Graduate School of Medicine, Kyoto University, Japan.

出版信息

Invest Ophthalmol Vis Sci. 1998 Feb;39(2):358-63.

PMID:9477994

Abstract

PURPOSE

To develop a highly sensitive method for in vivo quantitation of intravitreal peroxides by vitreous fluorophotometry with 2',7'-dichlorofluorescin (DCFH), a hydrogen peroxide (H2O2)-sensitive fluorescent dye, and to measure peroxides in the vitreous humor after panretinal laser photocoagulation (PRP).

METHODS

In the presence of H2O2 and lipid hydroperoxides, nonfluorescent DCFH was oxidized to highly fluorescent 2',7'-dichlorofluorescein (DCF; excitation, 495 nm; emission, 520 nm), which is detectable by vitreous fluorophotometry. Reactions of DCFH, including hematin with various concentrations of H2O2, were investigated in vivo. Fluorophotometry with DCFH was performed 1, 7, 14, and 28 days and 2 months after argon laser PRP. Untreated eyes served as the controls.

RESULTS

Exogenously applied H2O2 oxidized DCFH to DCF in a dose-dependent manner, ranging from 6 x 10(-8) mol/l to 6 x 10(-5) mol/l in concentration in vivo. Intravitreal DCF concentration was 83.7 +/- 6.8 nmol/l in control eyes. A significant increase of DCF was detected 1 day after PRP (330.7 +/- 123.8 nmol/l, P < 0.002). The increase peaked on day 7 (352.4 +/- 239.5 nmol/l, P < 0.002) and remained elevated at 2 months after PRP (161.8 +/- 51.4 nmol/l, P < 0.01).

CONCLUSIONS

This method allowed a highly sensitive quantitation of intravitreal peroxides in vivo. The authors' findings indicated that PRP induces increased production of peroxides in rabbit vitreous for 2 months. The data suggested that persistently high levels of peroxides in the vitreous humor affect the development of vitreous liquefaction after PRP.

摘要

目的

利用对过氧化氢（H2O2）敏感的荧光染料2'，7'-二氯荧光素（DCFH），通过玻璃体荧光光度法开发一种用于体内定量玻璃体内过氧化物的高灵敏度方法，并测量全视网膜激光光凝（PRP）后玻璃体液中的过氧化物。

方法

在H2O2和脂质氢过氧化物存在的情况下，非荧光性的DCFH被氧化为高荧光性的2'，7'-二氯荧光素（DCF；激发波长，495nm；发射波长，520nm），可通过玻璃体荧光光度法检测到。在体内研究了DCFH与包括不同浓度H2O2的血红素的反应。在氩激光PRP后1天、7天、14天、28天和2个月进行DCFH荧光光度法检测。未治疗的眼睛作为对照。

结果

在体内，外源性应用的H2O2以剂量依赖性方式将DCFH氧化为DCF，浓度范围为6×10(-8)mol/l至6×10(-5)mol/l。对照眼中玻璃体内DCF浓度为83.7±6.8nmol/l。PRP后1天检测到DCF显著增加（330.7±123.8nmol/l，P<0.002）。增加在第7天达到峰值（352.4±239.5nmol/l，P<0.002），并在PRP后2个月仍保持升高（161.8±51.4nmol/l，P<0.01）。