Li Lumin, Wang Baogui, Yu Ping, Wen Xuefang, Gong Deming, Zeng Zheling
State Key Laboratory of Food Science and Technology, Nanchang Univ, Nanchang, 330047, China.
School of Environmental and Chemical Engineering, Nanchang Univ, Nanchang, 330031, China.
J Food Sci. 2016 Jun;81(6):H1546-52. doi: 10.1111/1750-3841.13321. Epub 2016 May 3.
Medium chain fatty acids (MCFA) can be more easily absorbed and supply energy more rapidly than long chain fatty acids (LCFA). However, little is known about the inflammatory response by the treatment of MCFA in human liver cells. Thus this study used human liver cells (LO2) to evaluate the effects of MCFA on apoptosis and inflammatory response. Tetrazolim-based colorimetric assay and lactate dehydrogenase assay were used to measure the viability of LO2 cells, isolated spleens and liver cells from BALB/C mice. Inverted fluorescence microscopy and flow cytometry were used to assess the cell apoptosis. Activity of superoxide dismutase and malondialdehyde level were measured to determine the oxidative damage. mRNA or protein levels of classical pro-inflammatory cytokines were analyzed by quantitative real-time polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay and western blotting. The results showed that the liver cells treated with the fatty acids at 200 μM for 24 h exhibited good viability. Fatty acids induced inflammatory cytokines at transcriptional and translational levels to a lesser extent than lipopolysaccharide. LCFA (oleic acid) up-regulated tumor necrosis fator-α, monocyte chemoattractant-1 and interleukin-1β while down-regulated IL-6 and IL-8 secretion to a higher extent than MCFA in mRNA and protein levels. These findings suggested that MCFA may induce apoptosis to a less extent and exert more gentle inflammation than LCFA in human liver cells.
中链脂肪酸(MCFA)比长链脂肪酸(LCFA)更容易被吸收,且能更快地提供能量。然而,关于MCFA处理人肝细胞后的炎症反应,人们了解甚少。因此,本研究使用人肝细胞(LO2)来评估MCFA对细胞凋亡和炎症反应的影响。采用基于四氮唑的比色法和乳酸脱氢酶测定法来检测LO2细胞、从BALB/C小鼠分离的脾脏和肝细胞的活力。使用倒置荧光显微镜和流式细胞术评估细胞凋亡。通过测量超氧化物歧化酶活性和丙二醛水平来确定氧化损伤。通过定量实时聚合酶链反应(qPCR)、酶联免疫吸附测定和蛋白质印迹法分析经典促炎细胞因子的mRNA或蛋白质水平。结果表明,用200μM脂肪酸处理24小时的肝细胞具有良好的活力。与脂多糖相比,脂肪酸在转录和翻译水平上诱导炎症细胞因子的程度较低。在mRNA和蛋白质水平上,LCFA(油酸)上调肿瘤坏死因子-α、单核细胞趋化蛋白-1和白细胞介素-1β,同时下调IL-6和IL-8的分泌,其下调程度高于MCFA。这些发现表明,在人肝细胞中,MCFA诱导细胞凋亡的程度可能较小,且引发的炎症比LCFA更温和。