Patton G M, Clark S B, Fasulo J M, Robins S J
J Clin Invest. 1984 Jan;73(1):231-40. doi: 10.1172/JCI111196.
To determine the molecular species composition of lecithins of different nascent lipoproteins, high density lipoproteins (HDL), very low density lipoproteins (VLDL), and chylomicrons (CM) were isolated from the mesenteric lymph of rats. Lymph was collected at 0 degrees C with 5,5'-dithiobis-2-dinitrobenzoic acid added to inhibit lecithin-cholesterol acyl transferase. CM were separated by ultracentrifugation and HDL from VLDL by dextran SO4-MG+2 precipitation. Molecular species of lecithin were directly isolated by reverse phase high performance liquid chromatography. In fasted animals, the lecithin compositions of lymph HDL and VLDL were virtually the same and closely resembled the lecithin composition of intestinal mucosa. When bile lecithin was eliminated (by bile diversion), there was a marked change in lecithin composition of all lipoprotein and mucosal samples, which was most notable for a reduction in 16:0-species (which are predominant in bile) and a relative increase in the corresponding 18:0-species. Feeding unsaturated triglycerides (triolein, trilinolein, or a combination of triolein and trilinolein) also resulted in a change in HDL and VLDL lecithin composition. The effect was similar whether bile lecithin was present or eliminated and was notable for a reduction in 16:0-species, an increase in 18:0-species, and the emergence of large amounts of diunsaturated lecithins that corresponded to the fatty acid composition of the triglycerides fed (i.e., 18:1-18:1, 18:2-18:2, and 18:1-18:2 lecithins). When bile-diverted rats were infused via the duodenum with a mix of [14C]choline-labeled lecithins (isolated from the bile of other rats), the incorporation of infused lecithins into different lymph lipoproteins was distinctly different. Individual lecithins were incorporated to a variable extent into each lipoprotein. In fasted rats the specific activities of all major molecular species of lecithin were relatively greater in VLDL than HDL, indicating that HDL derived proportionately more of its lecithins from an endogenous pool than did VLDL. Feeding triolein changed the specific activities of more of the lecithin species of VLDL than of HDL. The specific activities of lecithins in CM were more similar to VLDL than to HDL after triolein feeding. Results thus indicate that, although the lecithins of different mesenteric lymph lipoproteins are similar and may be derived from membrane sites with the same lecithin composition, lecithins incorporated into different lipoproteins originate from different metabolic pools and/or by different mechanisms.
为了确定不同新生脂蛋白(高密度脂蛋白(HDL)、极低密度脂蛋白(VLDL)和乳糜微粒(CM))中卵磷脂的分子种类组成,从大鼠肠系膜淋巴中分离出这些脂蛋白。在0℃下收集淋巴液,并添加5,5'-二硫代双-2-二硝基苯甲酸以抑制卵磷脂-胆固醇酰基转移酶。通过超速离心分离CM,通过葡聚糖硫酸酯-Mg²⁺沉淀从VLDL中分离HDL。通过反相高效液相色谱直接分离卵磷脂的分子种类。在禁食动物中,淋巴HDL和VLDL的卵磷脂组成实际上相同,并且与肠黏膜的卵磷脂组成非常相似。当胆汁卵磷脂被去除(通过胆汁引流)时,所有脂蛋白和黏膜样品的卵磷脂组成都发生了显著变化,最明显的是16:0种类(在胆汁中占主导)减少,相应的18:0种类相对增加。喂食不饱和甘油三酯(三油精、三亚油酸甘油酯或三油精和三亚油酸甘油酯的组合)也导致HDL和VLDL卵磷脂组成发生变化。无论胆汁卵磷脂是否存在或被去除,效果都是相似的,表现为16:0种类减少、18:0种类增加以及大量对应于所喂食甘油三酯脂肪酸组成的二不饱和卵磷脂出现(即18:1-18:1、18:2-18:2和18:1-18:2卵磷脂)。当通过十二指肠向胆汁引流的大鼠输注从其他大鼠胆汁中分离出的[¹⁴C]胆碱标记的卵磷脂混合物时,输注的卵磷脂掺入不同淋巴脂蛋白的情况明显不同。各个卵磷脂掺入每种脂蛋白的程度各不相同。在禁食大鼠中,可以看到所有主要卵磷脂分子种类在VLDL中的比活性相对高于HDL,这表明HDL从内源性池中获得的卵磷脂比例比VLDL更大。喂食三油精改变了VLDL中比HDL中更多的卵磷脂种类的比活性。喂食三油精后,CM中卵磷脂的比活性与VLDL更相似,而与HDL不同。因此结果表明,尽管不同肠系膜淋巴脂蛋白中的卵磷脂相似,并且可能来自具有相同卵磷脂组成的膜部位,但掺入不同脂蛋白中的卵磷脂源自不同的代谢池和/或通过不同的机制产生。