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Fluorescence and ESR studies on membrane oxidative damage by gamma radiation

机译：荧光和ESR研究γ辐射对膜的氧化损伤

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Oxidative damage to cellular membranes critically controls the manifestation of cellular response to ionizing radiation. To gain further insight into the damaging mechanisms, we have investigated the effects of gamma-radiation-generated free-radical-mediated peroxidative damage in egg yolk lecithin unilamellar liposomal membranes by employing 1,6-diphenyl-1,3,5-hexatriene (DPH). Alterations in lipid bilayer fluidity and malondialdehyde (MDA) formation were measured in irradiated liposomal membranes as a function of radiation dose (0.1-1 kGy). A relationship seems to exist between the degree of radiation-induced peroxidative damage and the magnitude of DPH fluorescence decay in irradiated membranes. Radiation-induced membrane rigidization and MDA formation were significantly reduced when alpha-tocopherol, a natural membrane antioxidant, was present in the liposomes suggesting an involvement of lipid free radicals in the mechanism of the damage process. The results of the present study have been compared with those obtained by the electron spin resonance (ESR) technique on human erythrocyte ghost membranes with spin-labeled phospholipids having the unique capability to sensitively report on the dynamic state of the lipid environment inside the bilayer membrane. Iodoacetamide and N-ethylmaleimide spin labels were used to investigate alterations in membrane proteins. These results have contributed to our understanding of mechanisms involved in radiation membrane oxidative damage in terms of lipid peroxidation, fluidity changes and involvement of -SH groups of membrane proteins. Combined use of fluorescence and ESR spin-label techniques is of potential interest in probing the deeper molecular mechanisms of radiation injury in cellular membranes for developing strategies to modify the radiation damage to cells. [References: 17]
机译：对细胞膜的氧化损伤关键地控制了细胞对电离辐射的反应。为了进一步了解破坏机理，我们研究了通过使用1,6-二苯基-1,3,5-己三烯（1,6-二苯基-1,3,5-己三烯（g）辐射产生的自由基介导的蛋黄卵磷脂卵磷脂脂质体膜中的过氧化损伤。 DPH）。测量了脂质体膜的脂质双层流动性和丙二醛（MDA）形成的变化与辐射剂量（0.1-1 kGy）的关系。辐射引起的过氧化损伤的程度与被辐照膜中DPH荧光衰减的程度之间似乎存在关系。当脂质体中存在α-生育酚（一种天然的膜抗氧化剂）时，辐射诱导的膜僵化和MDA的形成显着降低，表明脂质自由基参与了损伤过程的机制。本研究的结果已与通过电子自旋共振（ESR）技术在人红细胞鬼膜上获得的结果进行了比较，该膜具有自旋标记的磷脂，具有独特的能力来敏感地报告双层膜内部脂质环境的动态状态。碘乙酰胺和N-乙基马来酰亚胺自旋标记用于研究膜蛋白的变化。这些结果有助于我们理解涉及脂质过氧化，流动性变化和膜蛋白的-SH基团参与的辐射膜氧化损伤的机理。荧光和ESR自旋标记技术的结合使用在探索细胞膜中辐射损伤的更深分子机制以开发策略来修饰对细胞的辐射损伤方面具有潜在的兴趣。 [参考：17]

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