Abstract：
Metabolism-associated fatty liver disease (MAFLD) is one of the most common causes of liver disease; however, the underlying processes remain unknown. This study aimed to investigate the changes of free fatty acids (FFA) on the expression of genes related to the AMP-activated protein kinase (AMPK) signaling pathway in the primary hepatocytes of laying hens. The primary hepatocytes of laying hens were treated with FFA (containing a 2:1 ratio of oleic and palmitic acids) for 24 h. FFA significantly increased lipid droplet accumulation, decreased glycogen synthesis, increased the levels of triglycerides (TG), total cholesterol (TC), reactive oxygen species (ROS), malondialdehyde (MDA), and glucose content in the supernatant (GLU) in the primary hepatocytes of laying hens, and decreased the levels of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD), as well as mitochondrial membrane potential (MMP). The results of the PCR array combined with Western blotting experiments showed that the activity of AMPK was inhibited. Inhibition of AMPK signaling pathway decreases the expression of genes involved in fatty acid oxidation, increases the expression of genes involved in lipid synthesis, decreases the expression of genes involved in glycogen synthesis, increases the expression of genes involved in glycolysis, increases the expression of genes involved in oxidative stress, and increases the expression of genes involved in cell proliferation and apoptosis. Taken together, our results suggest that FFA can affect the homeostasis of the AMPK signaling pathway by altering energy metabolic homeostasis, inducing oxidative stress, and adjusting the onset of cell proliferation and apoptosis.
摘要：
代谢相关脂肪性肝病(MAFLD)是肝病的最常见原因之一;然而,底层进程仍然未知。本研究旨在探讨游离脂肪酸(FFA)对蛋鸡原代肝细胞AMP激活蛋白激酶(AMPK)信号通路相关基因表达的影响。蛋鸡的原代肝细胞用FFA(含有2:1比例的油酸和棕榈酸)处理24小时。FFA显著增加脂滴积累，糖原合成减少，增加甘油三酯（TG）的水平，总胆固醇(TC),活性氧（ROS），丙二醛（MDA），蛋鸡原代肝细胞上清液（GLU）中的葡萄糖含量，总抗氧化能力(T-AOC)和超氧化物歧化酶(SOD)水平降低,以及线粒体膜电位（MMP）。PCR阵列结合Western印迹实验结果表明，AMPK的活性受到抑制。抑制AMPK信号通路降低脂肪酸氧化相关基因的表达,增加参与脂质合成的基因的表达，降低参与糖原合成的基因的表达，增加参与糖酵解的基因的表达，增加参与氧化应激的基因的表达，并增加参与细胞增殖和凋亡的基因的表达。一起来看,我们的结果表明，FFA可以通过改变能量代谢稳态来影响AMPK信号通路的稳态，诱导氧化应激，调节细胞增殖和凋亡的发生。
