Camins A, Diez-Fernandez C, Prieto P
Laboratory of Pharmacology and Pharmacognosy, Faculty of Pharmacy, University of Barcelona, 08028 Barcelona, Spain.
Toxicol In Vitro. 1999 Jun;13(3):437-43. doi: 10.1016/s0887-2333(99)00012-0.
The effect of oxidative stress induced by different concentrations of hydrogen peroxide on dog neutrophils was studied. This effect was measured using dichlorofluorescein-diacetate (DCFH-DA) and by the cell surface membrane expression of heat shock protein (HSP) 27kDa, HSP 72kDa and HSP 90kDa families. Hydrogen peroxide induced a concentration-dependent increase in DCFH oxidation (from 10(-6)mto 10(-4)m), and an increase in the cell surface expression of HSPs families. At a concentration of 10(-4)m, the percentage of positive cells that showed an oxidation of DCFH was 94.7%+/-5.2 (n=3). Only vitamin E (but not vitamin C) at a concentration of 0.5mm was able to inhibit the intracellular oxidative stress induced by hydrogen peroxide. The percentage of positive cells that express these proteins after the treatment with hydrogen peroxide (10(-4)m) was: 74%+/-3.5 for HSP 27, 72%+/-2.6 for HSP 72 and 73%+/-1.2 for HSP 90 (n=3). This cell surface expression was not abolished by either vitamin C or vitamin E. Localization of HSPs in plasma membrane is of immunological interest because they have been implicated in autoimmune diseases.
研究了不同浓度过氧化氢诱导的氧化应激对犬中性粒细胞的影响。使用二氯荧光素二乙酸酯（DCFH-DA）并通过热休克蛋白（HSP）27kDa、HSP 72kDa和HSP 90kDa家族的细胞表面膜表达来测量这种影响。过氧化氢诱导DCFH氧化呈浓度依赖性增加（从10^(-6)m到10^(-4)m），并且HSP家族的细胞表面表达增加。在10^(-4)m的浓度下，显示DCFH氧化的阳性细胞百分比为94.7%±5.2（n = 3）。仅浓度为0.5mm的维生素E（而非维生素C）能够抑制过氧化氢诱导的细胞内氧化应激。用过氧化氢（10^(-4)m）处理后表达这些蛋白质的阳性细胞百分比为：HSP 27为74%±3.5，HSP 72为72%±2.6，HSP 90为73%±1.2（n = 3）。这种细胞表面表达未被维生素C或维生素E消除。HSP在质膜中的定位具有免疫学意义，因为它们与自身免疫性疾病有关。
