Hart C M, Gupta M P, Evanoff V
Department of Medicine, Indiana University, Indianapolis, USA.
Exp Lung Res. 1997 Sep-Oct;23(5):405-25. doi: 10.3109/01902149709039235.
Altering the fatty acid composition of cultured porcine pulmonary artery endothelial cells (PAEC) modulates their susceptibility to oxidant stress. This study demonstrates that supplementing PAEC with oleic acid (18:1 omega 9), but not gamma-linolenic acid (18:3 omega 6), provided dose-dependent protection from hydrogen peroxide (H2O2)-induced cytotoxicity. It was hypothesized that 18:1 reduced PAEC susceptibility to oxidant stress by altering H2O2 metabolism. To test this hypothesis, confluent PAEC monolayers were treated with 100-200 microM H2O2 or control conditions 24 h after supplementation with 0.1 mM 18:1, 18:3, or vehicle for 3 h. Intracellular [H2O2] in control cells (14.4-29.0 pM), estimated from the rate of aminotriazole-mediated inactivation of endogenous catalase activity, increased following treatment with 200 microM H2O2 (19.0-37.3 pM). Supplementation with 18:1 attenuated increases in intracellular [H2O2] only in oxidant-exposed cells, whereas supplementation with 18:3 attenuated intracellular [H2O2] only in control cells. Supplementation with 18:1 or 18:3 tended to reduce or enhance PAEC lipid hydroperoxide content following H2O2 exposure, respectively, but did not alter PAEC reduced glutathione content, the activities of glutathione peroxidase or catalase, or H2O2 uptake and release. Alteration of H2O2 metabolism in cultured PAEC may contribute to the ability of fatty acids to modulate cellular oxidant susceptibility.
改变培养的猪肺动脉内皮细胞(PAEC)的脂肪酸组成可调节其对氧化应激的敏感性。本研究表明,用油酸(18:1 ω9)而非γ-亚麻酸(18:3 ω6)补充PAEC可提供剂量依赖性保护,使其免受过氧化氢(H2O2)诱导的细胞毒性。据推测,18:1通过改变H2O2代谢降低了PAEC对氧化应激的敏感性。为验证这一假设,在补充0.1 mM 18:1、18:3或溶剂3小时后24小时,将汇合的PAEC单层用100 - 200 μM H2O2处理或置于对照条件下。根据内源性过氧化氢酶活性的氨基三唑介导失活速率估算,对照细胞中的细胞内[H2O2](14.4 - 29.0 pM)在用200 μM H2O2处理后增加(19.0 - 37.3 pM)。仅在暴露于氧化剂的细胞中,补充18:1可减弱细胞内[H2O2]的增加,而仅在对照细胞中,补充18:3可减弱细胞内[H2O2]。分别在H2O2暴露后,补充18:1或18:3倾向于降低或增加PAEC脂质氢过氧化物含量,但不改变PAEC还原型谷胱甘肽含量、谷胱甘肽过氧化物酶或过氧化氢酶的活性,以及H2O2的摄取和释放。培养的PAEC中H2O2代谢的改变可能有助于脂肪酸调节细胞氧化应激敏感性的能力。