Department of Physiology, Faculty of Medicine and Nursing, University of the Basque Country UPV/EHU, Leioa, Spain.
Biocruces Bizkaia Health Research Institute, Barakaldo, Spain.
Methods Mol Biol. 2023;2675:15-26. doi: 10.1007/978-1-0716-3247-5_2.
The liver is a central organ in regulating the whole body metabolic homeostasis, and, among many other processes, it plays a crucial role in lipoprotein metabolism. The liver controls the secretion of very-low-density lipoproteins (VLDLs), particles specialized in the transport of liver lipids, mainly triglycerides (TGs), to the adipose tissue, heart, and muscle, among other tissues, providing fatty acids to be stored or to be used as an energy source. The analysis of this metabolic process provides relevant information about the crosstalk between the liver and other organs. It also helps to identify how the liver is able to secrete lipids to reduce its accumulation. This protocol shows how to analyze the liver TG secretion rate blocking the VLDL clearance from the blood by the administration of poloxamer 407. In addition, it shows how to isolate the VLDL produced by the liver at the end of the experiment, so that the apolipoprotein and lipid content and size can be measured. Using antisense oligonucleotides (ASOs) for silencing target proteins involved in metabolic diseases has emerged as a new promising therapeutic approach. Thus, the usage of ASOs has also been included in this protocol. As a conclusion, evaluation of TG secretion rate in mice provides key information to understand the organ crosstalk in metabolic diseases and the capacity of the liver to secrete lipids to blood.
肝脏是调节全身代谢稳态的中心器官，在许多其他过程中，它在脂蛋白代谢中起着至关重要的作用。肝脏控制极低密度脂蛋白（VLDLs）的分泌，VLDL 专门用于将肝脏脂质，主要是甘油三酯（TGs），运输到脂肪组织、心脏和肌肉等组织，为储存或用作能量来源提供脂肪酸。对这种代谢过程的分析提供了关于肝脏和其他器官之间串扰的相关信息。它还有助于确定肝脏如何能够分泌脂质以减少其积累。本方案展示了如何通过施用泊洛沙姆 407 来阻止 VLDL 从血液中清除，从而分析肝脏 TG 分泌率。此外，它还展示了如何在实验结束时分离肝脏产生的 VLDL，以便测量载脂蛋白和脂质的含量和大小。使用反义寡核苷酸（ASOs）沉默代谢疾病相关靶蛋白已成为一种新的有前途的治疗方法。因此，本方案中也包括了 ASOs 的使用。总之，评估小鼠的 TG 分泌率提供了关键信息，可帮助理解代谢疾病中的器官串扰以及肝脏向血液分泌脂质的能力。
