Reichelt Kristin, Niebisch Anna M, Kacza Johannes, Schoeniger Axel, Fuhrmann Herbert
Faculty of Veterinary Medicine, Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
BioImaging Core Facility, Faculty of Veterinary Medicine, University of Leipzig, Leipzig, Germany.
Front Vet Sci. 2022 Mar 11;9:840202. doi: 10.3389/fvets.2022.840202. eCollection 2022.
Hepatosteatosis is a common metabolic disorder of dairy cows, especially during early lactation. Currently, there are a few models of bovine hepatic steatosis available, including primary hepatocytes, liver slices, and animal models. Studies that elucidate the influence of single fatty acids on lipid classes, fatty acid pattern, gene expression, and phenotypic changes are still limited. Hence, we investigated the suitability of the fetal bovine hepatocyte-derived cell line BFH12 as a model for hepatosteatosis. To create a steatotic environment, we treated BFH12 with stearic acid, palmitic acid, or oleic acid in non-toxic doses. Thin-layer chromatography and gas chromatography were used to analyze lipid classes and fatty acid pattern, and qPCR was used to quantify gene expression of relevant target genes. Lipid droplets were visualized with confocal laser scanning microscopy and evaluated for number and size. Treatment with oleic acid increased triglycerides, as well as lipid droplet count per cell and upregulated carnitine palmitoyl transferase 1, which correlates with findings of models. Oleic acid was largely incorporated into triglycerides, phospholipids, and non-esterified fatty acids. Stearic acid was found mainly in non-esterified fatty acids and triglycerides, whereas palmitic acid was mainly desaturated to palmitoleic acid. All three fatty acids downregulated stearyl-CoA-desaturase 1. In conclusion, BFH12 can acquire a steatotic phenotype by incorporating and accumulating fatty acids. Oleic acid is particularly suitable to produce hepatosteatosis. Therefore, BFH12 may be a useful model to study bovine hepatosteatosis and its underlying molecular mechanisms.
肝脂肪变性是奶牛常见的一种代谢紊乱疾病，尤其是在泌乳早期。目前，有几种可用的牛肝脂肪变性模型，包括原代肝细胞、肝切片和动物模型。阐明单一脂肪酸对脂质类别、脂肪酸模式、基因表达和表型变化影响的研究仍然有限。因此，我们研究了胎牛肝细胞系BFH12作为肝脂肪变性模型的适用性。为了创建脂肪变性环境，我们用无毒剂量的硬脂酸、棕榈酸或油酸处理BFH12。采用薄层色谱法和气相色谱法分析脂质类别和脂肪酸模式，并用qPCR定量相关靶基因的基因表达。用共聚焦激光扫描显微镜观察脂滴，并对其数量和大小进行评估。油酸处理增加了甘油三酯以及每个细胞的脂滴数量，并上调了肉碱棕榈酰转移酶1，这与模型的研究结果相关。油酸大量掺入甘油三酯、磷脂和非酯化脂肪酸中。硬脂酸主要存在于非酯化脂肪酸和甘油三酯中，而棕榈酸主要去饱和为棕榈油酸。所有三种脂肪酸均下调硬脂酰辅酶A去饱和酶1。总之，BFH12可以通过摄取和积累脂肪酸获得脂肪变性表型。油酸特别适合于诱导肝脂肪变性。因此，BFH12可能是研究牛肝脂肪变性及其潜在分子机制的有用模型。
