Käkelä Reijo, Somerharju Pentti, Tyynelä Jaana
Institute of Biomedicine/Biochemistry, Biomedicum Helsinki, University of Helsinki, Haartmaninkatu 8, 00014 Helsinki, Finland.
J Neurochem. 2003 Mar;84(5):1051-65. doi: 10.1046/j.1471-4159.2003.01602.x.
Phospholipids (PL) in cerebral cortex from patients with infantile (INCL or CLN1) and juvenile (JNCL or CLN3) forms of neuronal ceroid-lipofuscinosis (NCL) and controls were analysed by normal phase HPLC and on-line electrospray ionization ion-trap mass spectrometric detection (LC-ESI-MS). The method provided quantitative data on numerous molecular species of different PL classes, which are not achieved by using the conventional chromatographic methods. Compared with the controls, the INCL brains contained proportionally more phosphatidylcholine (PC), and less phosphatidylethanolamine (PE) and phosphatidylserine (PS). Different molecular species of PC, PE, PS, phosphatidylinositol and sphingomyelin were quantified using multiple internal PL standards that differed in fatty acyl chain length and thus allowed correction for chain length dependency of instrument response. In INCL cortex, which had lost 65% of the normal PL content, the proportions of polyunsaturated molecular species, especially the PS and PE that contained docosahexaenoic acid (22:6n-3), were dramatically decreased. The membranes may have adapted to this alteration by increasing the proportions of PL molecules substituted with monounsaturated and short-chain fatty acids. Lysobisphosphatidic acid was highly elevated in the INCL brain and consisted mostly of polyunsaturated species. It is possible that changes in the composition of PL membranes accelerate progression of INCL by altering signalling and membrane trafficking in neurons.
采用正相高效液相色谱法（HPLC）和在线电喷雾电离离子阱质谱检测法（LC - ESI - MS），对患有婴儿型（婴儿神经轴索营养不良症，INCL或CLN1）和青少年型（青少年神经蜡样脂褐质沉积症，JNCL或CLN3）神经元蜡样脂褐质沉积症（NCL）的患者以及对照组患者大脑皮质中的磷脂（PL）进行了分析。该方法提供了不同磷脂类众多分子种类的定量数据，这是使用传统色谱方法无法实现的。与对照组相比，INCL患者的大脑中磷脂酰胆碱（PC）比例相对较高，而磷脂酰乙醇胺（PE）和磷脂酰丝氨酸（PS）比例较低。使用脂肪酸酰基链长度不同的多种内部磷脂标准品，对PC、PE、PS、磷脂酰肌醇和鞘磷脂的不同分子种类进行了定量，从而校正了仪器响应的链长依赖性。在PL含量丧失了65%的INCL皮质中，多不饱和分子种类的比例，尤其是含有二十二碳六烯酸（22:6n - 3）的PS和PE的比例，显著降低。细胞膜可能通过增加单不饱和脂肪酸和短链脂肪酸取代的PL分子比例来适应这种变化。溶血双磷脂酸在INCL患者大脑中高度升高，且主要由多不饱和种类组成。PL膜组成的变化可能通过改变神经元中的信号传导和膜运输来加速INCL的进展。
