In vitro effects of docosahexaenoic and eicosapentaenoic acid on human meibomian gland epithelial cells
In vitro effects of docosahexaenoic and eicosapentaenoic acid on human meibomian gland epithelial cells
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DOI：
10.1016/j.exer.2015.08.024
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发表时间：
2015-11-01
影响因子：
3.4
通讯作者：
Paulsen, Friedrich
中科院分区：
文献类型：
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作者：
Hampel, Ulrike;Krueger, Magret;Paulsen, Friedrich
探讨ω-3脂肪酸对体外培养的人睑板腺上皮细胞（HMGECs，细胞系）的影响。用二十二碳六烯酸（DHA）或与二十碳五烯酸（EPA）和乙酰唾液酸（阿萨）的组合刺激HMGEC。应用苏丹III脂肪染色、活力和增殖测定、电细胞基质阻抗传感、环氧合酶-2和15-脂氧合酶基因表达的实时PCR以及消退素D1（RvD 1）、IFN γ、TNF α和IL-6的ELISA。在血清培养的HMGECs中，在存在或不存在EPA的情况下，100 μ M DHA增加了脂滴积累和活力。相比之下，在无血清条件下用DHA和EPA培养的HMGEC显示出最小的脂质积累、降低的增殖和活力。当用DHA和EPA刺激时，无血清培养的HMGECs的归一化阻抗显著降低。与单独的DHA或EPA或对照相比，在含有血清的培养基中培养的HMGEC在DHA和EPA刺激下显示出增加的标准化阻抗。在用DHA和EPA处理72小时的HMGECs中，IL-6和IFN γ下调。一般而言，与含或不含DHA或EPA的血清培养基中的24小时相比，72小时后TNF α、IFN γ和IL-6水平降低。DHA处理后，RvD 1的浓度升高2倍。在DHA刺激72 h后，与对照组相比，环氧合酶-2基因表达降低。用DHA和阿萨治疗显示15-脂氧合酶基因表达降低，DHA孵育三天后该基因表达降低。DHA和EPA补充影响体外HMGEC，并通过影响细胞因子水平、降低考克斯-2表达和增加RvD 1的产生来支持抗炎作用。(C)2015爱思唯尔有限公司版权所有。
To investigate the effect of omega-3 fatty acids on human meibomian gland epithelial cells (HMGECs, cell line) in vitro. HMGECs were stimulated with docosahexaenoic acid (DHA) or combinations with eicosapentaenoic acid (EPA) and acetyl sialic acid (ASA). Sudan III fat staining, viability and proliferation assays, electric cell-substrate impedance sensing, real-time PCR for gene expression of cyclooxygenase-2 and 15-lipoxygenase and ELISAs for resolvin D1 (RvD1), IFN gamma, TNF alpha and IL-6 were applied. Lipid droplet accumulation and viability was increased by 100 mu M DHA in the presence or absence of EPA in serum cultured HMGECs. In contrast, HMGECs cultured with DHA and EPA under serum-free conditions showed minimal lipid accumulation, decreased proliferation and viability. Normalized impedance was significantly reduced in serum-free cultured HMGECs when stimulated with DHA and EPA. HMGECs cultured in serum containing medium showed increased normalized impedance under DHA and EPA stimulation compared to DHA or EPA alone or controls. IL-6 and IFN gamma were downregulated in HMGECs treated for 72 h with DHA and EPA. In general, TNF alpha, IFN gamma and IL-6 levels were decreased after 72 h compared to 24 h in serum containing medium with or without DHA or EPA. The concentration of RvD1 was elevated 2-fold after DHA treatment. Cyclooxygenase-2 gene expression decreased compared to controls during DHA stimulation after 72 h. Treatment with DHA and ASA revealed a decreased 15-lipoxygenase gene expression which was reduced after three days of DHA incubation. DHA and EPA supplementation affected HMGECs in vitro and supported anti-inflammatory effects by influencing cytokine levels, decreasing COX-2 expression and increasing the production of RvD1. (C) 2015 Elsevier Ltd. All rights reserved.