Hartbauer M, Hutter-Paie B, Windisch M
Institute of Zoology, Department of Neurobiology, Karl-Franzens University, Graz, Austria.
J Neural Transm (Vienna). 2001;108(5):581-92. doi: 10.1007/s007020170058.
Cerebrolysin (Cere, EBEWE Arzneimittel, Austria), a peptidergic drug produced by a standardised enzymatic breakdown of porcine brain proteins, consists of a mixture of 75% free amino acids and 25% low molecular weight peptides (<10 k DA). Cerebrolysin was shown to protect against MAP2 loss in primary embryonic chick neuronal cultures after brief histotoxic hypoxia and in a rat model of acute brain ischemia. Since MAP2 is involved in processes like neuronal growth, plasticity and dendritic branching, we address the question whether Cere is protecting processes against degeneration in a chronic low serum (2% FCS) cell stress model and whether the spontaneous outgrowth of axon-like processes is influenced. This was accomplished by quantification of the neurite lengths of embryonic chicken telencephalon neurons after 4 and 8 days. Additionally, time-lapse video microscopy was performed to study a possible influence of Cere on the growth cone behaviour of axon-like processes. To distinguish between effects caused by the peptide fraction and the effects related to free amino acids, we used an artificial amino acid solution (AA-mix). Results demonstrate a process outgrowth promoting effect of the AA-mix and Cere after 4 DIV. After 8 days neuronal network degeneration occurred in the AA-mix treated cultures, whereas Cere treated cultures still presented a well differentiated neuronal network. Dying neurons could release factors possibly impeding neurite outgrowth and Cere was shown to increase the viability of chicken cortical neurons. Neither the addition of BDNF nor serum supplementation (5% and 10% FCS) could protect the neuronal network against degeneration after 8 DIV, although these treatments were shown to ameliorate the viability of chicken telencephalon neurons. This result together with the finding obtained using the artificial amino acid solution points to the peptide fraction of Cere to be responsible for the protection of processes against degeneration. Time-lapse studies of Cere treated cultures revealed a significant decrease of the velocities characterising random growth cone movements, which is thought to be responsible for an increase in the length of axon-like processes after 4 DIV.
脑蛋白水解物（Cere，奥地利EBEWE制药公司生产）是一种通过对猪脑蛋白进行标准化酶解产生的肽能药物，由75%的游离氨基酸和25%的低分子量肽（<10 kDa）组成。研究表明，在短暂的组织毒性缺氧后，脑蛋白水解物可保护原代胚胎鸡神经元培养物中的微管相关蛋白2（MAP2）不丢失，在急性脑缺血大鼠模型中也有同样作用。由于MAP2参与神经元生长、可塑性和树突分支等过程，我们探讨了在慢性低血清（2%胎牛血清）细胞应激模型中，脑蛋白水解物是否能保护相关过程不发生退化，以及其对轴突样突起的自发生长是否有影响。这通过对4天和8天后胚胎鸡端脑神经元的神经突长度进行定量来实现。此外，进行了延时视频显微镜观察，以研究脑蛋白水解物对轴突样突起生长锥行为的可能影响。为了区分肽部分和游离氨基酸的作用，我们使用了一种人工氨基酸溶液（AA混合物）。结果表明，在接种后4天，AA混合物和脑蛋白水解物均有促进突起生长的作用。8天后，用AA混合物处理的培养物中出现神经元网络退化，而用脑蛋白水解物处理的培养物仍呈现出分化良好的神经元网络。垂死的神经元可能释放出可能阻碍神经突生长的因子，而脑蛋白水解物被证明可提高鸡皮质神经元的活力。尽管添加脑源性神经营因子（BDNF）和补充血清（5%和10%胎牛血清）在接种后8天均不能保护神经元网络不发生退化，但这些处理被证明可改善鸡端脑神经元的活力。这一结果与使用人工氨基酸溶液获得的发现共同表明，脑蛋白水解物的肽部分负责保护相关过程不发生退化。对用脑蛋白水解物处理的培养物进行的延时研究表明，表征随机生长锥运动的速度显著降低，这被认为是接种后4天轴突样突起长度增加的原因。
