Lipid peroxidation contributes to hydrogen peroxide induced cytotoxicity in renal epithelial cells
Lipid peroxidation contributes to hydrogen peroxide induced cytotoxicity in renal epithelial cells
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DOI：
10.1038/ki.1996.12
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发表时间：
1996-01-01
影响因子：
19.6
通讯作者：
Lieberthal, W
中科院分区：
文献类型：
--
作者：
Sheridan, AM;Fitzpatrick, S;Lieberthal, W
我们已经研究了脂质过氧化作用中的细胞毒性的过氧化氢在OK细胞含有显着不同数量的细胞膜多不饱和脂肪酸（PUFA）。在无血清培养基中生长的OK细胞中，检测不到PUFA。这些细胞的膜主要含有油酸、硬脂酸和棕榈酸。当在含有10%小牛血清的培养基中培养时，OK细胞含有可测量量的PUFA [亚油酸（5 +/- 1%）和花生四烯酸（8 +/- 1%）]。当含血清的培养基补充有60 mM亚油酸时，亚油酸（21 +/-1%）和花生四烯酸（15 +/-1%）的膜含量均显著增加。H2 O2诱导的OK细胞损伤的严重程度实质上改变了细胞膜的PUFA含量。将OK细胞暴露于1.25 mM H2 O2 1小时，导致在补充有PUFA含量“正常”的亚油酸的血清中生长的细胞（90 +/- 2%）比在未补充的小牛血清中生长的PUFA水平“降低”的细胞（81 +/- 3%）更多的细胞死亡（通过台盼蓝测定）。在含有不可检测水平的PUFA的确定的无血清培养基中生长的细胞遭受最少的H2 O2诱导的致死性细胞损伤（47 +/- 8%）。使用细胞活力的克隆形成试验，发现具有不同PUFA含量的细胞之间的H2 O2的细胞毒性的比较差异。1.25 mM H2 O2诱导的丙二醛（MDA）积累在PUFA含量“正常”的细胞（702 +/- 103 pM/100 μ g细胞DNA/hr）中比在PUFA含量“减少”的细胞（328 +/- 112 pM/100 μ g DNA/hr）中更大，在无血清培养基中生长的细胞中检测不到。总之，培养物中细胞的PUFA含量受培养条件的深刻影响。我们的数据提供了新的和直接的证据表明，细胞膜的过氧化直接导致H2 O2诱导的细胞损伤和死亡的严重程度。
We have examined the role of lipid peroxidation in the cytotoxicity of H2O2 in OK cells containing markedly differing amounts of cell membrane polyunsaturated fatty acids (PUFA). In OK cells grown in a serum free medium, PUFA were undetectable. The membranes of these cells contained predominantly oleic, stearic and palmitic acids. When cultured in medium containing 10% calf serum, OK cells contained measurable amounts of PUFA [linoleic (5 +/- 1%) and arachidonic acids (8 +/- 1%)]. When the serum containing medium was supplemented with 60 mM linoleic acid, the membrane content of both linoleic (21 +/- 1%) as well as arachidonic acid (15 +/- 1%) was substantially increased. The severity of injury induced by H2O2 in OK cells was substantially altered by the PUFA content of the cell membrane. Exposure of OK cells to 1.25 mM H2O2 for one hour resulted in more cell death (determined by a trypan blue assay) in cells grown in serum supplemented with linoleic acid with ''normal'' PUFA content (90 +/- 2%) than in cells with ''reduced'' levels of PUFA grown in unsupplemented calf serum (81 +/- 3%). Cells grown in defined, serum free medium with undetectable levels of PUFA suffered the least H2O2-induced lethal cell injury (47 +/- 8%). Comparable differences in the cytotoxicity of H2O2 among cells with differing PUFA content were found using a clonogenic assay of cell viability. Malondialdehyde (MDA) accumulation induced by 1.25 mM H2O2 was greater in cells with ''normal'' PUFA content (702 +/- 103 pM/100 mu g cell DNA/hr) than in cells with ''reduced'' PUFA (328 +/- 112 pM/100 mu g DNA/hr) and was undetectable in cells grown in defined, serum free medium. In summary, the content of PUFA of cells in culture is profoundly influenced by culture conditions. Our data provide novel and direct evidence that peroxidation of cell membranes contributes directly to the severity of cell injury and death induced by H2O2.