Slagsvold J E, Thorstensen K, Kvitland M, Mack M, Bjerve K S
Department of Medical Biochemistry, St. Olav's Hospital HF, Trondheim, Norway.
Scand J Clin Lab Invest. 2007;67(6):632-42. doi: 10.1080/00365510601175463.
The expression of delta 5 desaturase (D5D), delta 6 desaturase (D6D) and delta 9 desaturase (D9D) was determined by RT-PCR in the human promyelocytic cell line HL60. During 72 h of culture with 10% FBS, D5D and D6D were upregulated 5 to 6-fold, whereas D9D approximately doubled. The addition of fatty acids (FAs) to the culture medium suppressed upregulation of all desaturases. N-3 and n-6 FA appeared to be more effective than n-9 or saturated FA. When FAs were added after 72 h, further upregulation during the next 24 h was suppressed for nearly all desaturases and FAs tested, except for D5D when oleic acid (OA) or stearic acid (SA) was added. In cells cultured with restricted amounts of FBS, desaturase expression increased with decreasing concentrations of FBS. Cellular FA content decreased by 60% in the neutral lipid fraction, whereas that of the phospholipid fraction decreased by 10% during 72 h of culture. The largest decrease occurred in the sum of n-3 and n-6 FA of the neutral lipid fraction, which was reduced by 83%, whereas the content of these FAs in the phospholipid fraction decreased by 32%. The results indicate that when the supply of FA to HL60 cells is limited, the intracellular content of n-3 and n-6 FA decreases and this leads to upregulation of the desaturases, particularly D5D and D6D. Since HL60 cells resemble human leukocytes, the results suggest that desaturase expression in leukocytes may be exploited as a biomarker for FA status.
通过逆转录聚合酶链反应(RT-PCR)测定人早幼粒细胞系HL60中δ5去饱和酶(D5D)、δ6去饱和酶(D6D)和δ9去饱和酶(D9D)的表达。在用10%胎牛血清(FBS)培养72小时期间,D5D和D6D上调了5至6倍,而D9D大约增加了一倍。向培养基中添加脂肪酸(FAs)可抑制所有去饱和酶的上调。n-3和n-6脂肪酸似乎比n-9或饱和脂肪酸更有效。当在72小时后添加脂肪酸时,除添加油酸(OA)或硬脂酸(SA)时的D5D外,几乎所有测试的去饱和酶和脂肪酸在接下来的24小时内的进一步上调均受到抑制。在用有限量FBS培养的细胞中,去饱和酶表达随着FBS浓度的降低而增加。在72小时的培养过程中,中性脂质部分的细胞脂肪酸含量下降了60%,而磷脂部分的脂肪酸含量下降了10%。中性脂质部分中n-3和n-6脂肪酸的总和下降幅度最大,减少了83%,而磷脂部分中这些脂肪酸的含量下降了32%。结果表明,当HL60细胞的脂肪酸供应受到限制时,细胞内n-3和n-6脂肪酸的含量会下降,这会导致去饱和酶上调,尤其是D5D和D6D。由于HL60细胞类似于人类白细胞,结果表明白细胞中的去饱和酶表达可能被用作脂肪酸状态的生物标志物。