Ves-Losada A, Maté S M, Brenner R R
Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), Facultad de Ciencias Médicas, UNLP-CONICET, La Plata, Argentina.
Lipids. 2001 Mar;36(3):273-82. doi: 10.1007/s11745-001-0718-4.
Liver nuclear incorporation of stearic (18:0), linoleic (18:2n-6), and arachidonic (20:4n-6) acids was studied by incubation in vitro of the [1-14C] fatty acids with nuclei, with or without the cytosol fraction at different times. The [1-14C] fatty acids were incorporated into the nuclei as free fatty acids in the following order: 18:0 > 20:4n-6 >> 18:2n-6, and esterified into nuclear lipids by an acyl-CoA pathway. All [1-14C] fatty acids were esterified mainly to phospholipids and triacylglycerols and in a minor proportion to diacylglycerols. Only [1-14C]18:2n-6-CoA was incorporated into cholesterol esters. The incorporation was not modified by cytosol addition. The incorporation of 20:4n-6 into nuclear phosphatidylcholine (PC) pools was also studied by incubation of liver nuclei in vitro with [1-14C]20:4n-6-CoA, and nuclear labeled PC molecular species were determined. From the 15 PC nuclear molecular species determined, five were labeled with [1-14C]20:4n-6-CoA: 18:0-20:4, 16:0-20:4, 18:1-20:4, 18:2-20:4, and 20:4-20:4. The highest specific radioactivity was found in 20:4-20:4 PC, which is a minor species. In conclusion, liver cell nuclei possess the necessary enzymes to incorporate exogenous saturated and unsaturated fatty acids into lipids by an acyl-CoA pathway, showing specificity for each fatty acid. Liver cell nuclei also utilize exogenous 20:4n-6-CoA to synthesize the major molecular species of PC with 20:4n-6 at the sn-2 position. However, the most actively synthesized is 20:4-20:4 PC, which is a quantitatively minor component. The labeling pattern of 20:4-20:4 PC would indicate that this molecular species is synthesized mainly by the de novo pathway.
通过在不同时间将[1-14C]脂肪酸与细胞核在有或无胞质溶胶组分的情况下进行体外孵育,研究了硬脂酸(18:0)、亚油酸(18:2n-6)和花生四烯酸(20:4n-6)在肝细胞核中的掺入情况。[1-14C]脂肪酸以游离脂肪酸形式按以下顺序掺入细胞核:18:0>20:4n-6>>18:2n-6,并通过酰基辅酶A途径酯化为核脂质。所有[1-14C]脂肪酸主要酯化为磷脂和三酰甘油,少量酯化为二酰甘油。只有[1-14C]18:2n-6-辅酶A掺入胆固醇酯。添加胞质溶胶不改变掺入情况。还通过在体外将肝细胞核与[1-14C]20:4n-6-辅酶A孵育来研究20:4n-6掺入核磷脂酰胆碱(PC)库中的情况,并测定了核标记的PC分子种类。在所测定的15种PC核分子种类中,有5种被[1-14C]20:4n-6-辅酶A标记:18:0-20:4、16:0-20:4、18:1-20:4、18:2-20:4和20:4-20:4。在20:4-20:4 PC中发现了最高的比放射性,而这是一种次要种类。总之,肝细胞核拥有通过酰基辅酶A途径将外源性饱和和不饱和脂肪酸掺入脂质的必要酶,对每种脂肪酸具有特异性。肝细胞核还利用外源性20:4n-6-辅酶A在sn-2位合成含20:4n-6的PC主要分子种类。然而,最活跃合成的是20:4-20:4 PC,它在数量上是次要成分。20:4-20:4 PC的标记模式表明该分子种类主要通过从头合成途径合成。