Wirthensohn G, Lefrank S, Wirthensohn K, Guder W G
Biochim Biophys Acta. 1984 Sep 12;795(2):392-400. doi: 10.1016/0005-2760(84)90090-0.
Renal phospholipid metabolism was studied in rat cortical tubule suspensions by combining net measurements with precursor incorporation studies in order to be able to calculate the turnover of the different phospholipid species. Net amounts of tubular phosphatidylcholine (PC), phosphatidylinositol (PI) and phosphatidylethanolamine (PE) were 96.7 +/- 3.9, 29.2 +/- 2.9 and 74.1 +/- 6.0, mumol per g protein, respectively. Incubation of tubules in the absence or presence of renal substrates did not change net phospholipid contents. The predominant fatty acids were palmitate (47%) in PC and stearate in PI (56%) and PE (41%). Highest [14C]palmitate and [14C]glycerol incorporation rates were found in PC. In contrast, 32P-labeling was highest in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate exceeding PI-labeling by a factor of 2.4 and 5.2, respectively. In contrast, [3H]inositol incorporation into phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate was only 20% compared to that into PI. Addition of renal substrates like lactate, glutamine, glycerol and fatty acids increased precursor incorporation. The stimulating effects of gluconeogenic precursors and fatty acids were more than additive. Comparison of rates of [14C]glycerol, [32P]- and [3H]inositol incorporation suggests that de novo phospholipid biosynthesis is represented by these measurements, while the fatty acid fraction in addition turns over by exchange processes. According to our conclusions PE, PC, PI and phosphoinositides turn over with a half-life of 139, 16, 15 and 0.7 h, respectively, under optimal in vitro conditions.
通过将净测量与前体掺入研究相结合,对大鼠皮质肾小管悬浮液中的肾磷脂代谢进行了研究,以便能够计算不同磷脂种类的周转率。肾小管中磷脂酰胆碱(PC)、磷脂酰肌醇(PI)和磷脂酰乙醇胺(PE)的净含量分别为每克蛋白质96.7±3.9、29.2±2.9和74.1±6.0微摩尔。在不存在或存在肾底物的情况下孵育肾小管,不会改变磷脂的净含量。主要脂肪酸在PC中为棕榈酸(47%),在PI中为硬脂酸(56%),在PE中为硬脂酸(41%)。在PC中发现最高的[14C]棕榈酸和[14C]甘油掺入率。相比之下,磷脂酰肌醇4-磷酸和磷脂酰肌醇4,5-二磷酸中的32P标记最高,分别比PI标记高2.4倍和5.2倍。相比之下,与掺入PI相比,[3H]肌醇掺入磷脂酰肌醇4-磷酸和磷脂酰肌醇4,5-二磷酸的量仅为20%。添加乳酸、谷氨酰胺、甘油和脂肪酸等肾底物会增加前体掺入。糖异生前体和脂肪酸的刺激作用大于相加作用。[14C]甘油、[32P]和[3H]肌醇掺入率的比较表明,这些测量代表了从头合成磷脂的生物合成,而脂肪酸部分还通过交换过程进行周转。根据我们的结论,在最佳体外条件下,PE、PC、PI和磷酸肌醇的半衰期分别为139、16、15和0.7小时。