Chêne Gérald, Dubourdeau Marc, Balard Patricia, Escoubet-Lozach Laure, Orfila Claudine, Berry Antoine, Bernad José, Aries Marie-Françoise, Charveron Marie, Pipy Bernard
Macrophages, Mediateurs de l'Inflammation et Interactions Cellulaires, Université Paul Sabatier, EA 2405- INSERM IFR 31, Institut Louis Bugnard, BP 84225, 31432 Toulouse CEDEX 4, France.
Biochim Biophys Acta. 2007 May;1771(5):576-89. doi: 10.1016/j.bbalip.2007.02.014. Epub 2007 Mar 16.
Polyunsaturated fatty acids (PUFA) n-3 inhibit inflammation, in vivo and in vitro in keratinocytes. We examined in HaCaT keratinocyte cell line whether eicosapentaenoic acid (EPA) a n-3 PUFA, gamma-linoleic acid (GLA) a n-6 PUFA, and arachidic acid a saturated fatty acid, modulate expression of cyclooxygenase-2 (COX-2), an enzyme pivotal to skin inflammation and reparation. We demonstrate that only treatment of HaCaT with GLA and EPA or a PPARgamma ligand (roziglitazone), induced COX-2 expression (protein and mRNA). Moreover stimulation of COX-2 promoter activity was increased by those PUFAs or rosiglitazone. The inhibitory effects of GW9662 and T0070907 (PPARgamma antagonists), on COX-2 expression and on stimulation of COX-2 promoter activity by EPA and GLA suggest that PPARgamma is implicated in COX-2 induction. Finally, PLA2 inhibitor methyl arachidonyl fluorophosphonate blocked the PUFA effects on COX-2 induction, promoter activity and arachidonic acid mobilization suggesting involvement of AA metabolites in PPAR activation. These findings demonstrate that n-3 and n-6 PUFA increased PPARgamma activity is necessary for the COX-2 induction in HaCaT human keratinocyte cells. Given the anti-inflammatory properties of EPA, we suggest that induction of COX-2 in keratinocytes may be important in the anti-inflammatory and protective mechanism of action of PUFAs n-3 or n-6.
多不饱和脂肪酸(PUFA)n-3在体内和体外均可抑制角质形成细胞的炎症反应。我们在HaCaT角质形成细胞系中研究了n-3多不饱和脂肪酸二十碳五烯酸(EPA)、n-6多不饱和脂肪酸γ-亚麻酸(GLA)以及饱和脂肪酸花生酸是否能调节环氧合酶-2(COX-2)的表达,COX-2是一种在皮肤炎症和修复过程中起关键作用的酶。我们发现,仅用GLA、EPA或过氧化物酶体增殖物激活受体γ(PPARγ)配体(罗格列酮)处理HaCaT细胞,可诱导COX-2表达(蛋白质和mRNA水平)。此外,这些多不饱和脂肪酸或罗格列酮可增强COX-2启动子活性。GW9662和T0070907(PPARγ拮抗剂)对COX-2表达以及EPA和GLA对COX-2启动子活性的刺激作用具有抑制作用,这表明PPARγ参与了COX-2的诱导过程。最后,磷脂酶A2抑制剂甲基花生四烯酰氟磷酸酯可阻断多不饱和脂肪酸对COX-2诱导、启动子活性和花生四烯酸动员的影响,提示花生四烯酸代谢产物参与了PPAR激活。这些发现表明,n-3和n-6多不饱和脂肪酸增加PPARγ活性对于HaCaT人角质形成细胞中COX-2的诱导是必要的。鉴于EPA的抗炎特性,我们认为角质形成细胞中COX-2的诱导可能在n-3或n-6多不饱和脂肪酸的抗炎和保护作用机制中起重要作用。