Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo, 183-8509, Japan.
Vet Dermatol. 2020 Oct;31(5):419-e112. doi: 10.1111/vde.12881. Epub 2020 Aug 17.
Few studies have investigated the effects of essential fatty acids on the production of epidermal ceramide (CER) in canine keratinocytes.
HYPOTHESIS/OBJECTIVES: To investigate the effects of eicosapentaenoic acid (EPA) and linoleic acid (LA) supplementation on the production of CERs using an in vitro canine keratinocyte culture system.
Canine keratinocyte cells (MSCEK) were incubated with high Ca [1.8 mM calcium chloride (CaCl )] serum-free medium, supplemented with 3 µM EPA and 15 µM LA when the cells showed confluency. On Day 8 of application, lipid analysis using high-performance thin layer chromatography and real-time PCR for detecting glucosylceramide synthase and ceramidase were performed.
It was revealed that the amounts of CER (EOS) (combination of ω-hydroxy fatty acids and sphingosines), CER [EOP] (combination of ω-hydroxy fatty acids and phytosphingosines) and a mixture of CER [NS] (combination of nonhydroxy fatty acids and sphingosines) and [NDS] (combination of nonhydroxy fatty acids and dihydrosphingosines), as well as total CERs, were significantly increased in cells incubated with EPA and LA compared to those of the vehicle, with increased mRNA expression of glucosylceramide synthase.
These findings suggest that EPA and LA can potentially alter the CER profile of the skin and this may contribute to its epidermal barrier function in canine skin.
鲜有研究调查必需脂肪酸对犬角质形成细胞表皮神经酰胺（CER）产生的影响。
假说/目的：通过体外犬角质形成细胞培养系统，研究二十碳五烯酸（EPA）和亚油酸（LA）补充对 CER 产生的影响。
当细胞达到汇合状态时，用高钙[1.8 mM 氯化钙（CaCl ）]无血清培养基孵育犬角质形成细胞（MSCEK），并添加 3 µM EPA 和 15 µM LA。在应用的第 8 天，进行高效薄层色谱法的脂质分析和实时 PCR 检测葡萄糖神经酰胺合酶和神经酰胺酶。
结果表明，与载体相比，孵育 EPA 和 LA 的细胞中 CER（EOS）（ω-羟基脂肪酸和神经酰胺的组合）、CER [EOP]（ω-羟基脂肪酸和植物神经酰胺的组合）和 CER [NS]（非羟基脂肪酸和神经酰胺的组合）和[NDS]（非羟基脂肪酸和二氢神经酰胺的组合）以及总 CER 的量显著增加，葡萄糖神经酰胺合酶的 mRNA 表达增加。
这些发现表明，EPA 和 LA 可能改变皮肤的 CER 谱，这可能有助于其在犬皮肤中的表皮屏障功能。
