Montalto M B, Bensadoun A
Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853.
J Lipid Res. 1993 Mar;34(3):397-407.
Fatty acids (FA) were added to differentiating chick adipocytes to study their effects on the synthesis and secretion of avian lipoprotein lipase (LPL). Oleate (18:1n-9), eicosapentaenoate (EPA, 20:5n-3), and linoleate (18:2n-6) were complexed to fatty acid-free bovine serum albumin (BSA) and separately added to cells in RMPI-1640 media containing 0.5% delipidated hen serum. LPL secretion in the presence of 10 U/ml heparin was used as a means of estimating LPL synthesis. FA from 50 microM to 165 microM depressed LPL secretion for cells exposed to 20:5n-3 and 18:2n-6. The decrease in adipocyte LPL secretion was only observed with chronic administration of FA. In other experiments, the rate of LPL synthesis was estimated by incorporation of radiolabel (Tran 35S-label) in cells exposed to 50 microM FA. Total radioactivity incorporated into LPL, expressed as a percentage of total trichloroacetic acid (TCA)-precipitable radioactivity, was not affected in cells fed 18:1n-9 but was significantly decreased for cells fed 20:5n-3 (45%) or 18:2n-6 (67%) relative to controls given equimolar BSA (33 microM). Abundance of LPL message in similarly treated cells also decreased for cells incubated with 20:5n-3 or 18:2n-6 (35%) relative to controls and 18:1n-9-treated cells. A decrease in adipocyte LPL secretion was also observed with administration of lipoprotein (d < 1.006 g/ml) enriched in n-3 and n-6 fatty acids. LPL secretion of cells incubated with n-3 enriched lipoprotein at 50 microM and 75 microM triglyceride fatty acid equivalents was significantly greater than that of cells incubated with n-6-enriched lipoprotein. Interestingly, at the same concentrations of triglyceride fatty acids, lipoproteins enriched with n-9 fatty acids had no effect on LPL secretion relative to controls. These studies document that in cultured avian adipocytes, LPL secretion, synthesis, and level of message are decreased by chronic administration of n-3 and n-6 fatty acids. In contrast, in adipocytes supplemented with oleic acid there was no effect on LPL synthesis and secretion.
将脂肪酸(FA)添加到正在分化的鸡脂肪细胞中,以研究它们对禽脂蛋白脂肪酶(LPL)合成和分泌的影响。将油酸(18:1n-9)、二十碳五烯酸(EPA,20:5n-3)和亚油酸(18:2n-6)与无脂肪酸的牛血清白蛋白(BSA)复合,然后分别添加到含有0.5%脱脂鸡血清的RMPI-1640培养基中的细胞中。以10 U/ml肝素存在下的LPL分泌作为估计LPL合成的一种方法。对于暴露于20:5n-3和18:2n-6的细胞,50 microM至165 microM的FA会抑制LPL分泌。仅在长期给予FA时才观察到脂肪细胞LPL分泌减少。在其他实验中,通过将放射性标记(Tran 35S标记)掺入暴露于50 microM FA的细胞中来估计LPL合成速率。相对于给予等摩尔BSA(33 microM)的对照,掺入LPL中的总放射性,以总三氯乙酸(TCA)可沉淀放射性的百分比表示,在喂食18:1n-9的细胞中不受影响,但在喂食20:5n-3(45%)或18:2n-6(67%)的细胞中显著降低。与对照和用18:1n-9处理的细胞相比,用20:5n-3或18:2n-6孵育的细胞中,类似处理细胞中LPL信息的丰度也降低(35%)。给予富含n-3和n-6脂肪酸的脂蛋白时,也观察到脂肪细胞LPL分泌减少。在50 microM和75 microM甘油三酯脂肪酸当量下,用富含n-3的脂蛋白孵育的细胞的LPL分泌显著大于用富含n-6的脂蛋白孵育的细胞。有趣的是,在相同浓度的甘油三酯脂肪酸下,富含n-9脂肪酸的脂蛋白相对于对照对LPL分泌没有影响。这些研究证明,在培养的禽脂肪细胞中,长期给予n-3和n-6脂肪酸会降低LPL分泌、合成和信息水平。相比之下,在补充油酸的脂肪细胞中,对LPL合成和分泌没有影响。