Tkachenko Anton S, Klochkov Vladimir K, Lesovoy Vladimir N, Myasoedov Valeriy V, Kavok Nataliya S, Onishchenko Anatolii I, Yefimova Svetlana L, Posokhov Yevgen O
Kharkiv National Medical University, Nauky ave 4, Kharkiv, 61022, Ukraine.
Institute for Scintillation Materials, National Academy of Sciences of Ukraine, Kharkiv, Ukraine.
Wien Med Wochenschr. 2020 May;170(7-8):189-195. doi: 10.1007/s10354-020-00735-4. Epub 2020 Feb 12.
To assess the phospholipid bilayer of white blood cells (WBCs) and the ability of leukocytes to generate reactive oxygen species (ROS) in rats orally exposed to GdVO:Eu nanoparticle (VNP) solution for 2 weeks by fluorescent probes-ortho-hydroxy derivatives of 2,5-diaryl‑1,3‑oxazole.
Steady-state fluorescence spectroscopy, i.e., a study by the environment-sensitive fluorescent probes 2‑(2'-OH-phenyl)-5-(4'-phenyl-phenyl)-1,3-oxazole (probe O6O) and 2‑(2'-OH-phenyl)-phenanthro[9,10]-1,3-oxazole (probe PH7), and flow cytometry, i.e., analysis of 2',7'-dichlorofluorescein (DCF), a product of a dye 2',7'-dichlorodihydrofluorescein diacetate (HDCFDA), fluorescence in CD45/7-aminoactinomycin D (7-AAD) cells, were used to evaluate the state of cell membranes and reactive oxygen species (ROS) generation in leukocytes of rats orally exposed to gadolinium orthovanadate nanoparticles(VNPs).
No significant changes were detected in the spectra of the fluorescent probes bound to the WBCs from the rats orally exposed to nanoparticles in comparison with the corresponding spectra of the probes bound to the cells from the control group of animals. This indicates that in the case of the rats orally exposed to nanoparticles, no noticeable changes in physicochemical properties (i.e., in the polarity and the proton-donor ability) are observed in the lipid membranes of WBCs in the region where the probes locate. There was no statistically significant difference in the amount of ROS viable leukocytes in rats treated with VNPs and control samples.
Neither changes in the physical and chemical properties of the leukocyte membranes nor in ROS generation by WBCs are detected in the rats orally exposed to VNP solution for 2 weeks.
通过荧光探针——2,5-二芳基-1,3-恶唑的邻羟基衍生物,评估经口暴露于钒酸钆:铕纳米颗粒(VNP)溶液2周的大鼠白细胞的磷脂双层以及白细胞产生活性氧(ROS)的能力。
采用稳态荧光光谱法,即通过环境敏感型荧光探针2-(2'-羟基苯基)-5-(4'-苯基苯基)-1,3-恶唑(探针O6O)和2-(2'-羟基苯基)-菲并[9,10]-1,3-恶唑(探针PH7)进行研究,以及流式细胞术,即分析经口暴露于钒酸钆纳米颗粒(VNPs)的大鼠白细胞中2',7'-二氯荧光素(DCF)(一种染料2',7'-二氯二氢荧光素二乙酸酯(HDCFDA)的产物)在CD45/7-氨基放线菌素D(7-AAD)细胞中的荧光,来评估细胞膜状态和白细胞中产生活性氧(ROS)的情况。
与对照组动物细胞上结合的探针相应光谱相比,经口暴露于纳米颗粒的大鼠白细胞上结合的荧光探针光谱未检测到显著变化。这表明,在经口暴露于纳米颗粒的大鼠中,在探针所处区域的白细胞脂质膜中,未观察到物理化学性质(即极性和质子供体能力)有明显变化。经VNPs处理的大鼠与对照样品中,活性白细胞中ROS的量没有统计学上的显著差异。
经口暴露于VNP溶液2周的大鼠,未检测到白细胞膜的物理化学性质变化,也未检测到白细胞产生活性氧的变化。