Alvaro D, Cantafora A, Gandin C, Masella R, Santini M T, Angelico M
II Department of Gastroenterology, University of Rome La Sapienza, Italy.
Biochim Biophys Acta. 1989 Nov 6;1006(1):116-20. doi: 10.1016/0005-2760(89)90331-7.
The content of polyunsaturated phosphatidylcholines (PCs) is one of the parameters which regulate membrane functions. Polyunsaturated PCs are preferentially synthesized in the liver by the microsomal enzyme phosphatidylethanolamine N-methyltransferase. The activity of this enzyme may be stimulated in vitro in isolated rat hepatocytes by supplementation with dimethylethanolamine (DME), the polar head group of the precursor of PC along this pathway. The aim of this study was to evaluate in vivo the effect of an intravenous infusion of DME in the rat on the hepatic phospholipid composition. Bile fistula rats were intravenously infused for 15 h with sodium taurocholate (1 mumol/kg per min), with or without the addition of 0.3 mg/kg per min of [14C]DME. The concentration per gram of wet liver of individual phospholipid classes, PC molecular species and of total triacylglycerols, as well as the distribution of radioactivity in liver phospholipids, in rat tissues and body fluids were analyzed. A significant (P less than 0.01) enrichment in PC was found in the liver of DME-infused rats with respect to controls. No differences in the other phospholipid classes were found. DME-infused rats showed a significant (P less than 0.01) decrease in the hepatic concentration of triacylglycerols. At HPLC analysis, the enrichment in PC in DME-infused rats was found to be selectively due to three molecular species (i.e., sn-1 stearoyl/sn-2 arachidonoyl, sn-1 stearoyl/sn-2 linoleoyl, sn-1 stearoyl/sn-2 docosahexanoyl molecular species). In agreement with quantitative data, more than 70% of hepatic radioactivity was recovered in polyunsaturated PC species, with the highest specific activity in the sn-1 stearoyl PCs. The specific activity of hepatic PC approximates that of phosphatidyldimethylethanolamine. This finding together with the effective incorporation of DME in PC suggests that this amino base is methylated after its incorporation into phosphatidyldimethylethanolamine, throughout the stimulation of hepatic N-methyltransferase activity. The selective hepatic enrichment with polyunsaturated PC species after DME infusion may offer a new experimental tool for studying hepatic membrane metabolism.
多不饱和磷脂酰胆碱(PCs)的含量是调节膜功能的参数之一。多不饱和PCs优先在肝脏中由微粒体酶磷脂酰乙醇胺N-甲基转移酶合成。在离体大鼠肝细胞中,通过补充二甲基乙醇胺(DME)(该途径中PC前体的极性头部基团)可在体外刺激该酶的活性。本研究的目的是在体内评估静脉输注DME对大鼠肝脏磷脂组成的影响。对胆瘘大鼠静脉输注牛磺胆酸钠(1 μmol/kg每分钟)15小时,添加或不添加0.3 mg/kg每分钟的[14C]DME。分析了每克湿肝中各个磷脂类、PC分子种类以及总三酰甘油的浓度,以及大鼠组织和体液中肝脏磷脂的放射性分布。与对照组相比,输注DME的大鼠肝脏中PC有显著(P<0.01)富集。其他磷脂类未发现差异。输注DME的大鼠肝脏中三酰甘油浓度显著(P<0.01)降低。在高效液相色谱分析中,发现输注DME的大鼠肝脏中PC的富集选择性地归因于三种分子种类(即sn-1硬脂酰/sn-2花生四烯酰、sn-1硬脂酰/sn-2亚油酰、sn-1硬脂酰/sn-2二十二碳六烯酰分子种类)。与定量数据一致,超过70%的肝脏放射性在多不饱和PC种类中回收,sn-1硬脂酰PC的比活性最高。肝脏PC的比活性接近磷脂酰二甲基乙醇胺的比活性。这一发现连同DME有效掺入PC表明,该氨基酸在掺入磷脂酰二甲基乙醇胺后通过刺激肝脏N-甲基转移酶活性而被甲基化。DME输注后肝脏中多不饱和PC种类的选择性富集可能为研究肝膜代谢提供一种新的实验工具。