Pompella A, Comporti M
Istituto di Patologia Generale, Università, Siena, Italy.
Am J Pathol. 1993 May;142(5):1353-7.
Confocal laser scanning fluorescence microscopy plus image videoanalysis was used to visualize the tissue areas and the subcellular sites first involved by oxidative stress and lipid peroxidation, in the well-established experimental model of lipid peroxidation induced by haloalkane intoxication in the liver cell. The fluorescent reagent 3-hydroxy-2-naphthoic acid hydrazide was employed to derivativize the carbonyl functions originating from the lipoperoxidative process in situ, in liver cryostat sections from in vivo intoxicated rats, as well as in isolated hepatocytes exposed in vitro to the pro-oxidant action of haloalkanes. The results obtained indicate that: 1) the detection of fluorescent derivatives of carbonyls indeed offers a gain in sensitivity, 2) haloalkane-induced lipid peroxidation in hepatocytes primarily involves the perinuclear endoplasmic reticulum, whereas the plasma membrane and the nuclear compartment are unaffected, and 3) lipid peroxidation also induces an increase of liver autofluorescence.
在已建立的肝细胞中卤代烷中毒诱导脂质过氧化的实验模型中，共聚焦激光扫描荧光显微镜结合图像视频分析用于观察首先受到氧化应激和脂质过氧化影响的组织区域和亚细胞位点。荧光试剂3-羟基-2-萘甲酰肼用于原位衍生化来自脂质过氧化过程的羰基官能团，这些羰基官能团存在于体内中毒大鼠的肝脏冷冻切片中，以及体外暴露于卤代烷促氧化作用的分离肝细胞中。获得的结果表明：1）羰基荧光衍生物的检测确实提高了灵敏度；2）卤代烷诱导的肝细胞脂质过氧化主要涉及核周内质网，而质膜和核区室未受影响；3）脂质过氧化还导致肝脏自发荧光增加。
