Stable transfection of fatty acid translocase (CD36) in a rat heart muscle cell line (H9c2).
Stable transfection of fatty acid translocase (CD36) in a rat heart muscle cell line (H9c2).
复制标题
在大鼠心肌细胞系 (H9c2) 中稳定转染脂肪酸转位酶 (CD36)。
作者：
F. V. van Nieuwenhoven;J. Luiken;Y. D. de Jong;P. Grimaldi;G. J. van der Vusse;J. Glatz
脂肪酸转位酶 (FAT/CD36) 是一种膜蛋白，推测参与长链脂肪酸的跨膜转运。我们测试了这样的假设：这种蛋白在 H9c2（一种通常不表达 FAT 的大鼠心脏细胞系）中表达会增加细胞棕榈酸的摄取。我们能够用 FAT 稳定转染 H9c2 细胞，产生 15 个显示不同水平 FAT 表达的细胞系。首先在未转染的 H9c2 细胞和两种 FAT 转染的细胞系中研究了棕榈酸的摄取和代谢。在每种情况下，棕榈酸酯的吸收在至少 30 分钟内随时间呈线性，并且随着棕榈酸酯浓度的增加，吸收率达到饱和。在细胞内棕榈酸酯处理的最大能力未充分利用的条件下，我们测试了 15 个 FAT 转染细胞系中的 7 个，具有不同的 FAT 表达水平。 FAT 表达水平与棕榈酸酯摄取率之间没有发现显着相关性。总之，我们发现H9c2细胞对棕榈酸酯的摄取主要通过被动扩散发生。脂肪酸转位酶 (FAT) 转染并没有显着增加棕榈酸酯摄取率，这增加了 H9c2 细胞缺乏一种蛋白质（或一组蛋白质）的可能性，该蛋白质在长链脂肪酸从细胞外区室转运到细胞质的过程中充当 FAT 的必需伙伴。
Fatty acid translocase (FAT/CD36) is a membrane protein putatively involved in the transmembrane transport of long-chain fatty acids. We tested the hypothesis that expression of this protein in H9c2, a rat heart cell line normally not expressing FAT, would increase cellular palmitate uptake. We were able to stably transfect H9c2 cells with FAT, yielding 15 cell lines showing varying levels of FAT expression. The uptake and metabolism of palmitate was first studied in the non-transfected H9c2 cells and in two FAT-transfected cell lines. In each case, uptake of palmitate was found to be linear in time for at least 30 min and the uptake rate was saturable with increasing palmitate concentrations. Using conditions under which the maximal capacity of intracellular palmitate handling was not fully utilized, we tested 7 out of 15 FAT-transfected cell lines with varying FAT expression levels. No significant correlation was found between the level of FAT expression and the rate of palmitate uptake. In conclusion, we found that palmitate uptake by H9c2 cells occurs mainly by passive diffusion. Fatty acid translocase (FAT) transfection did not significantly increase the palmitate uptake rate, raising the possibility that H9c2 cells lack a protein (or set of proteins) that acts as an obligatory partner of FAT in long-chain fatty acid transport from the extracellular compartment to the cytoplasm.
登录
查看更多内容
DOI：
10.1016/s0021-9258(17)47247-4
发表时间：
1984-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
N. Abumrad;Jane H. Park;Charles R. Park
通讯作者：
N. Abumrad;Jane H. Park;Charles R. Park
影响因子：
20.3
作者：
D. Greenwalt;Robert Lipsky;C. Ockenhouse;H. Ikeda;N. Tandon;G. Jamieson
通讯作者：
D. Greenwalt;Robert Lipsky;C. Ockenhouse;H. Ikeda;N. Tandon;G. Jamieson
DOI：
10.1073/pnas.93.7.2646
发表时间：
1996-04-02
影响因子：
11.1
作者：
Ibrahimi, A;Sfeir, Z;Abumrad, NA
通讯作者：
Abumrad, NA
DOI：
10.1073/pnas.87.9.3484
发表时间：
1990-05-01
影响因子：
11.1
作者：
BERK, PD;WADA, H;THUNG, S
通讯作者：
THUNG, S
影响因子：
20.1
作者：
SIPIDO, KR;MARBAN, E
通讯作者：
MARBAN, E