Suyatna F D, van Veldhoven P P, Borgers M, Mannaerts G P
Afdeling Farmakologie, Katholieke Universiteit Leuven, Belgium.
J Mol Cell Cardiol. 1988 Jan;20(1):47-62. doi: 10.1016/s0022-2828(88)80178-0.
Sarcolemmal vesicles were purified to a similar extent, 50- to 60-fold on a protein basis, from normal rat hearts and hearts subjected to 30 or 60 min of autolysis at 37 degrees C (total ischemia in vitro). Electron microscopic examination of the autolytic hearts revealed sarcolemmal discontinuities and other morphological characteristics typical of irreversible cell injury. Total contents and percentage composition of phospholipid classes did not differ between normal and autolytic hearts or between sarcolemmal preparations from these hearts. There was no increase in lysophospholipid contents of whole hearts or of purified sarcolemma after autolysis. Long chain acyl-CoAs or acylcarnitines did not accumulate in autolytic hearts under our experimental conditions. The molar long chain acyl-CoA: phospholipid ratio in isolated sarcolemma was extremely low (1:100,000). It increased 3-fold after autolysis but the increase was most probably the result of an increase in mitochondrial contamination of the sarcolemmal preparations from autolytic hearts. The molar long chain acylcarnitine: phospholipid ratio of isolated sarcolemma was much larger (1:100), but it did not change after autolysis. Experiments, in which radioactive amphiphiles were incorporated in isolated sarcolemma that was subsequently repeatedly washed, indicated that the lysophospholipid and acylcarnitine contents of isolated sarcolemma reflect the contents of sarcolemma in situ, but that sarcolemmal acyl-CoA is used for re-acylation reactions during purification, explaining the low acyl-CoA content of isolated sarcolemma. Na/K-ATPase and Na/Ca-exchange activities were markedly depressed in isolated sarcolemma from autolytic hearts. Our results suggest that sarcolemmal phospholipid breakdown and sarcolemmal amphiphile accumulation are not responsible for the structural and functional defects of the sarcolemma after autolysis.
从正常大鼠心脏以及在37℃下经历30或60分钟自溶(体外完全缺血)的心脏中,肌膜囊泡在蛋白质基础上被纯化到相似的程度,即50至60倍。对自溶心脏的电子显微镜检查显示肌膜连续性中断以及其他不可逆细胞损伤的典型形态特征。正常心脏和自溶心脏之间或来自这些心脏的肌膜制剂之间,磷脂类别的总含量和百分比组成没有差异。自溶后,全心脏或纯化肌膜的溶血磷脂含量没有增加。在我们的实验条件下,长链酰基辅酶A或酰基肉碱在自溶心脏中没有积累。分离的肌膜中长链酰基辅酶A与磷脂的摩尔比极低(1:100,000)。自溶后它增加了3倍,但这种增加很可能是由于自溶心脏的肌膜制剂中线粒体污染增加的结果。分离的肌膜中长链酰基肉碱与磷脂的摩尔比要大得多(1:100),但自溶后没有变化。将放射性两亲物掺入随后反复洗涤的分离肌膜中的实验表明,分离肌膜的溶血磷脂和酰基肉碱含量反映了原位肌膜的含量,但肌膜酰基辅酶A在纯化过程中用于再酰化反应,这解释了分离肌膜中酰基辅酶A含量低的原因。自溶心脏分离的肌膜中钠钾ATP酶和钠钙交换活性明显降低。我们的结果表明,肌膜磷脂分解和肌膜两亲物积累与自溶后肌膜的结构和功能缺陷无关。