Shrestha Nirajan, Cuffe James S M, Holland Olivia J, Perkins Anthony V, McAinch Andrew J, Hryciw Deanne H
School of Medical Science, Griffith University, Southport, Australia.
Institute for Health and Sport, Victoria University, Melbourne, Australia.
Cell Physiol Biochem. 2019;52(1):94-108. doi: 10.33594/000000007. Epub 2019 Feb 18.
BACKGROUND/AIMS: The omega 6 fatty acid (FA) linoleic acid (LA) is required for embryonic development; however, omega 6 FAs can alter cellular metabolism via inflammation or modulation of mitochondrial function. Fetal LA is obtained from the maternal diet, and FAs are transported to the fetus via placental FA transporters (FATPs) and binding proteins (FABPs), but specific proteins responsible for LA transport in placental trophoblasts are unknown. Dietary LA consumption is increasing, but the effect of elevated LA on trophoblast function is not clear.
Swan71 trophoblasts were exposed to physiological and supraphysiological concentrations of LA for 24 hours. Quantification of mRNA was determined using real time PCR, and protein concentration was determined by Western blot analysis. Cell viability, citrate synthase activity and mitochondrial respiration were determined.
Exposure to 300 and 500 μM LA increased FATP1 and FATP4 mRNA expression. 500 μM LA increased FATP1 and FATP4 protein expression. Exposure to 500 μM increased FABP5 mRNA expression, while exposure to 100 to 500 μM LA decreased FABP3 mRNA expression. 300 and 500 μM LA decreased FABP3 protein expression. Cell viability was decreased by exposure to LA (100 to 1000 μM). Citrate synthase activity and routine mitochondrial respiration were significantly decreased by exposure to 300 and 500 μM LA, and maximal respiration and spare respiratory capacity were decreased by exposure to 100 to 500 μM LA. 300 and 500 μM LA increased reactive oxygen species generation in human trophoblasts. Moreover, exposure to 300 and 500 μM LA decreased IL-6 secretion. Exposure to 500 μM LA increased IL-8, NF-κB and PPAR-γ mRNA expression, but decreased NF-κB protein expression. 300 μM LA decreased IL-8 protein expression. Further, exposure to 100 to 500 μM LA increased prostaglandin E2 and leukotriene B₄ release.
Exposure to LA decreases cell viability, alters mRNA expression of FA transport related proteins, mitochondrial respiration and function, and inflammatory responses in trophoblasts. These findings may have implications on placental function when women consume high levels of LA.
背景/目的：ω-6脂肪酸（FA）亚油酸（LA）是胚胎发育所必需的；然而，ω-6脂肪酸可通过炎症或线粒体功能调节来改变细胞代谢。胎儿的LA来自母体饮食，脂肪酸通过胎盘脂肪酸转运蛋白（FATP）和结合蛋白（FABP）转运至胎儿，但胎盘滋养层细胞中负责LA转运的特定蛋白尚不清楚。膳食中LA的摄入量在增加，但其对滋养层细胞功能的影响尚不清楚。
将Swan71滋养层细胞暴露于生理浓度和超生理浓度的LA中24小时。使用实时PCR定量mRNA，通过蛋白质印迹分析测定蛋白质浓度。测定细胞活力、柠檬酸合酶活性和线粒体呼吸。
暴露于300和500μM的LA可增加FATP1和FATP4 mRNA表达。500μM的LA可增加FATP1和FATP4蛋白表达。暴露于500μM可增加FABP5 mRNA表达，而暴露于100至500μM的LA可降低FABP3 mRNA表达。300和500μM的LA可降低FABP3蛋白表达。暴露于LA（100至1000μM）会降低细胞活力。暴露于300和500μM的LA会显著降低柠檬酸合酶活性和常规线粒体呼吸，暴露于100至500μM的LA会降低最大呼吸和备用呼吸能力。300和500μM的LA可增加人滋养层细胞中的活性氧生成。此外，暴露于300和500μM的LA会降低IL-6分泌。暴露于500μM的LA会增加IL-8、NF-κB和PPAR-γ mRNA表达，但会降低NF-κB蛋白表达。300μM的LA会降低IL-8蛋白表达。此外，暴露于100至500μM的LA会增加前列腺素E2和白三烯B₄的释放。
暴露于LA会降低细胞活力，改变FA转运相关蛋白的mRNA表达、线粒体呼吸和功能以及滋养层细胞中的炎症反应。当女性摄入高水平的LA时，这些发现可能对胎盘功能有影响。
