Schürer N Y, Rippke F, Vogelsang K, Schliep V, Ruzicka T
Department of Dermatology, University of Düsseldorf, Germany.
Arch Dermatol Res. 1999 Jan;291(1):47-53. doi: 10.1007/s004030050382.
Epidermal linoleic acid, i.e. essential fatty acid (EFA), is essential for cutaneous barrier function. Cultured human keratinocytes, routinely used for studies of lipid metabolism, are grown in a keratinocyte serum-free medium (KSFM), under conditions that reveal EFA-deficient cells. Here, fatty acid (FA) uptake was analysed in human adult keratinocytes grown either under EFA-deficient conditions [KSFM supplemented with 10% FCS (A) or 1% UltroserG (B)] or EFA-supplemented conditions [KSFM supplemented with a devised FA cocktail (C) or evening primrose oil (D)]. The FA composition of the total cellular lipid and major lipid fractions was analysed by gas chromatography. Cells grown with supplements A or B balanced their EFA-deficient state primarily with oleic acid. Cells grown with supplements C or D normalized to the epidermal FA composition in vivo with raised linoleic and lower oleic acid contents. When cells were grown longer than 48 h with supplements C or D decreased cell growth was observed. FA uptake was curvilinear with preference for linoleic over oleic acid under all culture conditions. The uptake of linoleic acid by cells cultured with supplement B was twice the uptake of those cultured with supplement A, while the uptake of oleic acid was similar under both culture conditions. Oleic acid uptake of cells cultured with supplement C or D was lower. These results show that the uptake of linoleic, but not that of oleic acid, is influenced by the extracellular FA composition, and that EFA-supplemented keratinocytes compared to EFA-deficient cells might serve as an in vitro model for the study of EFA metabolism.
表皮亚油酸,即必需脂肪酸(EFA),对皮肤屏障功能至关重要。常用于脂质代谢研究的培养人角质形成细胞,在无血清角质形成细胞培养基(KSFM)中生长,处于揭示必需脂肪酸缺乏细胞的条件下。在此,分析了在必需脂肪酸缺乏条件下[补充10%胎牛血清(A)或1%优思格森(B)的KSFM]或补充必需脂肪酸条件下[补充设计的脂肪酸混合物(C)或月见草油(D)的KSFM]培养的成人角质形成细胞中的脂肪酸(FA)摄取情况。通过气相色谱分析总细胞脂质和主要脂质组分的脂肪酸组成。用补充剂A或B培养的细胞主要用油酸平衡其必需脂肪酸缺乏状态。用补充剂C或D培养的细胞,其体内的亚油酸含量升高,油酸含量降低,从而使表皮脂肪酸组成正常化。当细胞用补充剂C或D培养超过48小时时,观察到细胞生长下降。在所有培养条件下,脂肪酸摄取呈曲线状,对亚油酸的偏好高于油酸。用补充剂B培养的细胞对亚油酸的摄取是用补充剂A培养的细胞的两倍,而在两种培养条件下油酸的摄取相似。用补充剂C或D培养的细胞对油酸的摄取较低。这些结果表明,亚油酸而非油酸的摄取受细胞外脂肪酸组成的影响,并且与必需脂肪酸缺乏的细胞相比,补充必需脂肪酸的角质形成细胞可能作为研究必需脂肪酸代谢的体外模型。