Schoen Alexandra N, Weinrauch Alyssa M, Bouyoucos Ian A, Anderson W Gary
Department of Biological Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.
Bamfield Marine Sciences Centre, Bamfield, British Columbia, Canada.
Am J Physiol Regul Integr Comp Physiol. 2023 Nov 1;325(5):R534-R545. doi: 10.1152/ajpregu.00132.2023. Epub 2023 Aug 21.
The liver is an essential energy storage organ in vertebrates. In teleosts and elasmobranchs, previous studies examining hepatic energy balance have used isolated hepatocytes. Although these studies have been informative, the high-fat content in the elasmobranch liver limits isolation of hepatocytes and therefore the utility of this method to understand hepatic metabolic processes. In the present study, we developed an in situ liver perfusion in the North Pacific spiny dogfish . Perfusions were conducted by cannulating the hepatic portal vein (inflowing cannulation) and the sinus venosus through the heart (outflowing cannulation). Changes in major elasmobranch metabolites (glucose and 3-hydroxybutarate [3-HB]) were determined by the arterial (inflow)-venous (outflow) difference in metabolite concentration. Liver preparations were considered viable due to consistent oxygen consumption over 3 h and the maintenance of predictable vasoconstriction following administration of homologous 10 M angiotensin II (ANG II). Removal and reintroduction from the perfusate of metabolites showed endogenous 3-HB production in the isolated perfused livers but did not affect glucose balance. However, the arterial-venous difference of both metabolites did not change following perfusion with heterologous insulin and homologous glucagon, which may be due to the glucose intolerant nature of elasmobranchs. Ultimately, we show the viability of this perfusion for the investigation of hepatic energy mobilization in sharks. We describe a viable liver perfusion in a shark species for the first time as determined by oxygen consumption and hormone-mediated changes in hemodynamics (angiotensin II, ANG II). In addition, removal of major energy metabolites confirms hepatic ketone [3-hydroxybutyrate (3-HB)] production by an elasmobranch liver. Perfusion with heterologous insulin and homologous glucagon did not cause changes in glucose balance, however, possibly demonstrating differences in glucose metabolism in this taxon as compared with more derived vertebrates.
肝脏是脊椎动物重要的能量储存器官。在硬骨鱼和软骨鱼中,以往研究肝脏能量平衡时使用的是分离的肝细胞。尽管这些研究提供了很多信息,但软骨鱼肝中高脂肪含量限制了肝细胞的分离,因此这种方法在理解肝脏代谢过程方面的实用性受到限制。在本研究中,我们开发了一种北太平洋刺鲨的原位肝脏灌注方法。灌注通过将肝门静脉插管(流入插管)和经心脏的静脉窦插管(流出插管)进行。主要软骨鱼代谢物(葡萄糖和3 - 羟基丁酸酯[3 - HB])的变化通过代谢物浓度的动脉(流入)-静脉(流出)差异来确定。肝脏制剂被认为是有活力的,因为在3小时内氧气消耗一致,并且在给予同源10 M血管紧张素II(ANG II)后维持了可预测的血管收缩。代谢物从灌注液中去除和重新引入显示,在分离的灌注肝脏中有内源性3 - HB产生,但不影响葡萄糖平衡。然而,在用异源胰岛素和同源胰高血糖素灌注后,两种代谢物的动静脉差异没有变化,这可能是由于软骨鱼对葡萄糖不耐受的特性。最终,我们展示了这种灌注方法在研究鲨鱼肝脏能量动员方面的可行性。我们首次描述了一种基于氧气消耗和激素介导的血流动力学变化(血管紧张素II,ANG II)确定的鲨鱼肝脏灌注方法,该方法具有可行性。此外,去除主要能量代谢物证实了软骨鱼肝可产生肝酮[3 - 羟基丁酸(3 - HB)]。然而,用异源胰岛素和同源胰高血糖素灌注并没有导致葡萄糖平衡的变化,这可能表明与更进化的脊椎动物相比,该分类群在葡萄糖代谢方面存在差异。