Shiva Shankar Reddy C S, Subramanyam M V V, Vani R, Asha Devi S
Laboratory of Gerontology, Department of Zoology, Bangalore University, Bangalore 560 056, India.
Toxicol In Vitro. 2007 Dec;21(8):1355-64. doi: 10.1016/j.tiv.2007.06.010. Epub 2007 Jun 30.
The present study was designed to induce oxidative stress in lipid and aqueous phases through azo bis(2-amidinopropane)dihydrochloride (AAPH), 2,2'-azobis 2,4-dimethylvaleronitrile (ADVN) and hydrogen peroxide (H(2)O(2)) either alone or in combination with vitamin C or vitamin El and to assess the vulnerability of rat erythrocytes to oxidative stress. While AAPH acted equally on cell membrane and cytosol, ADVN increased OS in the membrane. The extent of hemolysis and increased membrane fragility caused was more in the case of azo compounds than of H(2)O(2). While vitamin E (2mM) reduced oxidative stress in the membrane, vitamin C (60mM) was more effective in the lysates. The concentration of malondialdehyde and advanced oxidation protein products was lowered by antioxidants. The level of lipofuscin, a product of lipid peroxidation was also increased by ADVN and H(2)O(2). Antioxidants, did, however, reduce the accumulation of protein carbonyl content in cells exposed to azo compounds although they were ineffective in inhibiting oxidation of membrane band 3 protein and sulphydryl content. Taken together, our study demonstrated the antioxidative property of vitamin E and vitamin C in reducing oxidative stress in aqueous as well as lipid phases of erythrocytes and further suggested the feasibility of in vitro models in evaluating the mechanisms of oxidative injury.
本研究旨在通过偶氮双（2-脒基丙烷）二盐酸盐（AAPH）、2,2'-偶氮双（2,4-二甲基戊腈）（ADVN）和过氧化氢（H₂O₂）单独或与维生素C或维生素E联合使用，在脂质相和水相中诱导氧化应激，并评估大鼠红细胞对氧化应激的易感性。AAPH对细胞膜和细胞质溶胶的作用相同，而ADVN增加了膜中的氧化应激。偶氮化合物引起的溶血程度和膜脆性增加比H₂O₂更明显。虽然维生素E（2mM）降低了膜中的氧化应激，但维生素C（60mM）在裂解物中更有效。抗氧化剂降低了丙二醛和晚期氧化蛋白产物的浓度。脂褐素（脂质过氧化产物）的水平也因ADVN和H₂O₂而增加。然而，抗氧化剂确实减少了暴露于偶氮化合物的细胞中蛋白质羰基含量的积累，尽管它们在抑制膜带3蛋白的氧化和巯基含量方面无效。综上所述，我们的研究证明了维生素E和维生素C在降低红细胞水相和脂质相中氧化应激方面的抗氧化特性，并进一步表明了体外模型在评估氧化损伤机制方面的可行性。
