Dontsov Alexander, Koromyslova Anna, Ostrovsky Mikhail, Sakina Natalia
Alexander Dontsov, Anna Koromyslova, Mikhail Ostrovsky, Natalia Sakina, Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 119334 Moscow, Russia.
World J Exp Med. 2016 Nov 20;6(4):63-71. doi: 10.5493/wjem.v6.i4.63.
To investigate the effect of two ways of lipofuscin production (lipid peroxidation and glycation) on lipofuscin fluorescence characteristics and phototoxicity and to compare them with the properties of natural lipofuscin.
Model lipofuscins were prepared on the basis of bovine photoreceptor outer segments (POS) with bisretinoid A2E addition. One set of samples was prepared from POS modified by lipid peroxidation, while another set from POS modified by glycation with fructose. Fluorescent properties and kinetics of photoinduced superoxide generation of model lipofuscins and human retinal pigment epithelium (RPE) lipofuscin were compared. The fluorescence spectra of samples were measured at 365 nm excitation wavelength and 380-650 emission wavelength.
The fluorescence spectra of model lipofuscins are almost the same as the spectrum of natural lipofuscin. Visible light irradiation of both model lipofuscins and natural lipofuscin isolated from RPE cells leads to decrease of a fluorescence maximum at 550 nm and to appearance of a distinct, new maximum at 445-460 nm. The rate of photogeneration of reactive oxygen forms by both model lipofuscins was almost the same and approximately two times less than that of RPE lipofuscin granules.
These data suggest that fluorescent characteristics and phototoxicity of lipofuscin granules depend only to an insignificant degree on the oxidative modification of POS proteins and lipids, and generally are defined by the bisretinoid fluorophores contained in them.
研究两种脂褐素生成方式(脂质过氧化和糖基化)对脂褐素荧光特性和光毒性的影响,并将其与天然脂褐素的特性进行比较。
在添加双视黄醛A2E的牛光感受器外段(POS)基础上制备模型脂褐素。一组样品由经脂质过氧化修饰的POS制备,另一组由经果糖糖基化修饰的POS制备。比较模型脂褐素和人视网膜色素上皮(RPE)脂褐素的荧光特性及光诱导超氧化物生成动力学。在365 nm激发波长和380 - 650 nm发射波长下测量样品的荧光光谱。
模型脂褐素的荧光光谱与天然脂褐素的光谱几乎相同。对模型脂褐素和从RPE细胞分离出的天然脂褐素进行可见光照射,均导致550 nm处荧光最大值降低,并在445 - 460 nm处出现明显的新最大值。两种模型脂褐素产生活性氧形式的光生成速率几乎相同,且约为RPE脂褐素颗粒的两倍。
这些数据表明,脂褐素颗粒的荧光特性和光毒性仅在很小程度上取决于POS蛋白和脂质的氧化修饰,总体上由其中所含的双视黄醛荧光团决定。