Márquez Martín Ana, de la Puerta Vázquez Rocío, Fernández-Arche Angeles, Ruiz-Gutiérrez Valentina
Instituto de la Grasa, C.S.I.C., Padre García Tejero S/N, Seville 41012, Spain.
Free Radic Res. 2006 Mar;40(3):295-302. doi: 10.1080/10715760500467935.
The pentacyclic triterpene maslinic acid (MA) is a natural compound present in the non glyceride fraction of pomace olive oil, also called orujo olive oil. This compound has previously demonstrated antioxidant properties against lipid peroxidation in vitro, but its effects on reactive oxygen and nitrogen-derived species and pro-inflammatory cytokines generated by a cell system have not yet been investigated. In this study, we have tested the effect of MA upon oxidative stress and cytokine production using peritoneal murine macrophages. MA significantly inhibited the enhanced production of nitric oxide (NO) induced by lypopolysaccharide (LPS) when it was measured by the nitrite production with an inhibitory concentration 50% value (IC(50)) of 25.4 microM. This inhibiting effect seems to be consequence of an action at the level of the LPS-induction of the inducible nitric oxide synthethase (iNOS) gene enzyme expression rather than to a direct inhibitory action on enzyme activity. The secretion of the inflammatory cytokines interleukine-6 and TNF-a from LPS-stimulated murine macrophages was also significantly reduced (p < 0.05 and 0.01) by 50 and 100 microM of MA. In addition, reactive oxygen species were measured after stimulation with phorbol-12-myristate-13-acetate (PMA). Thus, pre-treatment with MA reduced the generation of hydrogen peroxide from stimulated macrophages in a dose-dependent manner (IC(50): 43.6 microM) as assayed by the oxidation of the peroxidase enzyme. However, no inhibitory effect on superoxide release, measured by the reduction of ferricytochrome c, was observed after the pretreatment with MA in the culture medium. These results suggest a potential biopharmaceutical use of this hydroxy-pentacyclic triterpene derivative, present in orujo olive oil, on preventing oxidative stress and pro-inflammatory cytokine generation.
五环三萜马斯里酸（MA）是一种天然化合物，存在于果渣橄榄油（也称为orujo橄榄油）的非甘油酯部分。该化合物先前已在体外证明对脂质过氧化具有抗氧化特性，但尚未研究其对细胞系统产生的活性氧和氮衍生物种以及促炎细胞因子的影响。在本研究中，我们使用腹膜小鼠巨噬细胞测试了MA对氧化应激和细胞因子产生的影响。当通过亚硝酸盐产生测定时，MA以25.4 microM的50%抑制浓度（IC50）显著抑制了脂多糖（LPS）诱导的一氧化氮（NO）产量增加。这种抑制作用似乎是由于在诱导型一氧化氮合酶（iNOS）基因酶表达的LPS诱导水平上的作用，而不是对酶活性的直接抑制作用。50和100 microM的MA也显著降低了LPS刺激的小鼠巨噬细胞中炎性细胞因子白细胞介素-6和TNF-α的分泌（p < 0.05和0.01）。此外，在用佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）刺激后测量了活性氧物种。因此，如通过过氧化物酶的氧化测定，MA预处理以剂量依赖方式（IC50：43.6 microM）减少了刺激的巨噬细胞中过氧化氢的产生。然而，在用MA预处理培养基后，未观察到对通过铁细胞色素c还原测量的超氧化物释放的抑制作用。这些结果表明，这种存在于orujo橄榄油中的羟基五环三萜衍生物在预防氧化应激和促炎细胞因子产生方面具有潜在的生物制药用途。
