Hwang Jinhee, Singh Neetu, Long Charles, Smith Stephen B
Department of Animal Science, Texas A & M University, College Station, 2471 TAMU, TX 77843, USA.
Department of Veterinary Physiology and Pharmacology, Texas A & M University, College Station, 4466 TAMU, TX, 77843, USA.
Lipids. 2018 Oct;53(10):933-945. doi: 10.1002/lipd.12102.
The most highly regulated and abundant fatty acid in animal tissue is oleic acid (18:1n9). Oleic acid is synthesized by the Δ9 desaturase, stearoyl-CoA desaturase-1 (SCD1), which is responsible for the synthesis of the putative cytokine palmitoleic acid (16:1n7) and 18:2 cis-9, trans-11 conjugated linoleic acid. Owing to the importance of SCD1 in lipid metabolism, we generated porcine swine kidney (SK6) transgenic cell lines for sustained overexpression or knockdown of porcine stearoyl-CoA desaturase-1 (pSCD1) in an inducible manner by utilizing a lentiviral expression system. We successfully validated these cell culture models for expression and functionality of pSCD1 by documenting that the pSCD-transduced cells overexpressed pSCD1 protein and mRNA. Additionally, the pSCD1-transduced cells increased the conversion of palmitate (16:0) to palmitoleic acid nearly fourfold. The lentiviral vectors utilized in this study can be further used to generate transgenic animals to document the effects of the overexpression of SCD1 on obesity and steatosis.
动物组织中调控最为严格且含量最为丰富的脂肪酸是油酸(18:1n9)。油酸由Δ9去饱和酶——硬脂酰辅酶A去饱和酶-1(SCD1)合成,该酶负责合成假定的细胞因子棕榈油酸(16:1n7)以及18:2顺-9,反-11共轭亚油酸。鉴于SCD1在脂质代谢中的重要性,我们利用慢病毒表达系统,以可诱导的方式构建了猪肾(SK6)转基因细胞系,用于持续过表达或敲低猪硬脂酰辅酶A去饱和酶-1(pSCD1)。通过记录pSCD转导的细胞过表达pSCD1蛋白和mRNA,我们成功验证了这些细胞培养模型中pSCD1的表达及功能。此外,pSCD1转导的细胞将棕榈酸(16:0)转化为棕榈油酸的能力提高了近四倍。本研究中使用的慢病毒载体可进一步用于培育转基因动物,以证明SCD1过表达对肥胖和脂肪变性的影响。