Vouldoukis Ioannis, Lacan Dominique, Kamate Caroline, Coste Philippe, Calenda Alphonse, Mazier Dominique, Conti Marc, Dugas Bernard
Isocell Nutra SAS, Paris, France.
J Ethnopharmacol. 2004 Sep;94(1):67-75. doi: 10.1016/j.jep.2004.04.023.
The present study was conducted to evaluate in vitro and in vivo the antioxidant and anti-inflammatory properties of a cantaloupe melon (Cucumis melo LC., Cucurbitaceae) extract (CME) selected for its high superoxide dismutase activity. Peritoneal macrophages were pre-activated in vitro with 300 IU of interferon-gamma (IFN-gamma) and were then challenged in culture with IgGl/anti-IgG1 immune complexes (IgG1IC) in presence of various CME extracts. The subsequent production of free radicals (superoxide anion, nitric oxide, and peroxynitrite) and of pro-(TNF-alpha) and anti-(IL-10) inflammatory cytokines was evaluated. The CME inhibited in a dose-dependent manner the production of superoxide anion with a maximal effect at 100 microg/ml. This inhibitory effect of CME appeared to be closely linked to the SOD activity because it was dramatically decreased after heat inactivation of the SOD activity (HI-CME). In addition, the CME inhibited the production of peroxynitrite strengthening the antioxidant properties of this CME rich in SOD activity. The production of the pro- and anti-inflammatory cytokines, namely TNF-alpha and IL-10, being conditioned by the redox status of macrophages we also evaluated the effect of CME and HI-CME on the IgG1IC-induced cytokine production. When the SOD activity was present in the CME it promoted the IgG1IC-induced production of IL-10 instead of TNF-alpha. These data demonstrated that, in addition to its antioxidant properties, the anti-inflammatory properties of the CME extract were principally related to its capacity to induce the production of IL-10 by peritoneal macrophages. The particular properties of wheat gliadin (Triticum vulgare, Poaceae) for the oral delivery of functional proteins led us to test it in a new nutraceutical formula based on its combination with the CME thus monitoring the SOD activity release during the gastro-intestinal digestive process. In these experiments C57BL/6 mice were supplemented orally everyday during 28 days with: (1) the placebo, (2) the CME extract alone, (3) the gliadin, (4) the CME/gliadin combination, or (5) the HI-CME/gliadin combination (SOD inactivated). At the end of the supplementation period all the animals were injected intra-peritoneal (i.p.) with the pro-inflammatory cytokine IFN-gamma (300 IU) and peritoneal macrophages were harvested 24 h after to test their capacities to produce free radicals, TNF-alpha and IL-10 after triggering with IgG1IC. We demonstrated that animals supplemented during 28 days with the CME/gliadin combination were protected against the pro-inflammatory properties of IFN-gamma while the other products were inefficient. These data did not only indicate that the SOD activity is important for the antioxidant and anti-inflammatory properties of the CME extract, but also demonstrated that when the SOD activity is preserved during the digestive process by its combination with wheat gliadin it is possible to elicit in vivo the pharmacological effects of this antioxidant enzyme.
本研究旨在体外和体内评估一种因其高超氧化物歧化酶活性而被选中的哈密瓜(甜瓜属,葫芦科)提取物(CME)的抗氧化和抗炎特性。腹膜巨噬细胞在体外先用300 IU的γ-干扰素(IFN-γ)进行预激活,然后在各种CME提取物存在的情况下,用IgG1/抗IgG1免疫复合物(IgG1IC)在培养中进行刺激。随后评估自由基(超氧阴离子、一氧化氮和过氧亚硝酸盐)以及促炎(TNF-α)和抗炎(IL-10)细胞因子的产生。CME以剂量依赖的方式抑制超氧阴离子的产生,在100 μg/ml时具有最大效应。CME的这种抑制作用似乎与超氧化物歧化酶活性密切相关,因为在超氧化物歧化酶活性热失活(HI-CME)后,这种抑制作用显著降低。此外,CME抑制过氧亚硝酸盐的产生,增强了这种富含超氧化物歧化酶活性的CME的抗氧化特性。促炎和抗炎细胞因子,即TNF-α和IL-10的产生,受巨噬细胞氧化还原状态的调节,因此我们也评估了CME和HI-CME对IgG1IC诱导的细胞因子产生的影响。当CME中存在超氧化物歧化酶活性时,它促进IgG1IC诱导的IL-10产生而不是TNF-α。这些数据表明,除了其抗氧化特性外,CME提取物的抗炎特性主要与其诱导腹膜巨噬细胞产生IL-10的能力有关。小麦醇溶蛋白(普通小麦,禾本科)用于口服递送功能蛋白的特殊特性,使我们基于其与CME的组合在一种新的营养配方中对其进行测试,从而监测胃肠道消化过程中超氧化物歧化酶活性的释放。在这些实验中,C57BL/6小鼠在28天内每天口服补充:(1)安慰剂,(2)单独的CME提取物,(3)醇溶蛋白,(4)CME/醇溶蛋白组合,或(5)HI-CME/醇溶蛋白组合(超氧化物歧化酶失活)。在补充期结束时,所有动物腹腔内注射促炎细胞因子IFN-γ(300 IU),24小时后收集腹膜巨噬细胞,以测试它们在用IgG1IC触发后产生自由基、TNF-α和IL-10的能力。我们证明,在28天内用CME/醇溶蛋白组合补充的动物对IFN-γ的促炎特性具有保护作用,而其他产品则无效。这些数据不仅表明超氧化物歧化酶活性对CME提取物的抗氧化和抗炎特性很重要,而且还证明,当超氧化物歧化酶活性在消化过程中通过与小麦醇溶蛋白结合而得以保留时,有可能在体内引发这种抗氧化酶的药理作用。