The analysis of onset of familial amyotrophic lateral sclerosis(FALS) with His46Arg point mutation

研究了具有DNA断裂活性的糖基产物脱氢吡嗪(DHPs)与神经功能障碍的关系。His46Arg点突变FALS患者脊髓前角细胞内存在大量游离铜离子，DHPs的作用加速，中枢神经细胞似受损。为了验证这一假说，利用神经细胞系分析了DHPs对细胞功能的影响，并分析了神经细胞PC12和Schwann细胞中功能分子的变化。在具有不同DNA断裂活性的三种DHP中，断裂活性最高的DHP(Yl-3)对细胞功能有显著影响。Yl-3作用后雪旺细胞结构蛋白表达基本不变，但β-连环蛋白表达降低。虽然DHPS的加入引起的形态变化可能表明细胞凋亡…更严重的是，未检测到细胞凋亡标记分子PARP的断裂。低浓度DHP和铜离子单独存在时，细胞效应不明显。MAPK分子(ERK1/2、JNK和p38)在DHP和铜离子共存的雪旺细胞中被激活。提示低浓度DHP在体内可引起铜离子存在下的神经细胞变性。在细胞周期调控系统中，p27和p16的表达降低，cdc2蛋白失活。DHP处理的细胞周期可能表明细胞向G1/S方向收敛，推测有丝分裂过程受到抑制。这些结果表明，DHP在铜离子存在的情况下影响MAPK的活性，并增加了神经功能障碍的发生几率。另一方面，DHP与Cu2+的反应表明，Cu2+的存在加速了DHPS在反应体系中的作用。一部分结果已经在印刷中(Biol.Pharm.Bull，2005)，其他的已经提交，然后正在接受审查。较少

The correlation between neurological disorders and dehydropyrazines(DHPs) that sugar-derived product having the activity of DNA fragmentation was examined. The damage of SOD function bring the existence of many free copper ion, and the effects of DHPs were accelerated in the cells in anterior horn of spinal cord of FALS patient with His46Arg point mutation, thus cells in central nerve seemed to be impaired. To demonstrate this hypothesis, the affect on the cellular functions of DHPs was analyzed using nerve cell lines.The fluctuation of functional molecules in neural cell ; PC12 and Schwann cell, was assayed. Among three DHPs with different DNA fragmentation activity, DHP with the highest fragmentation activity (Yl-3) markedly affect the cellular function. The expression of structural proteins in the cells was almost unchanged, but beta-catenin expression was decreased in Schwann cells treated wjth Yl-3. Though the morphological changes by the addition of DHPs might indicate apoptosis … More occurred, the fragmentation of apoptosis marker molecule PARP was not detected.The cellular effect was not detected in the presence of low density of DHP and copper ion, independently. MAPK molecules (ERK1/2,JNK, and p38) activate in Schwann cells with coexistence of DHP and copper ion. This suggested that low density DHP could degenerate neural cells with the presence of copper ion in vivo. Among cell cycle regulation systems, the expression of p27 and p16 was decreased and cdc2 protein was inactivated. The cell cycle of DHP-treated cells might indicate the convergence into the G1/S state, and it postulated that mitosis process was inhibited.These results suggested that DHPs influenced MAPK activities with the presence of copper ion, and elevated the probability of onset of neurological disorder.On the other hand, the reaction of DHPs with Cu2+ demonstrated that the presence of Cu2+ accelerated the effect of DHPs in the reaction system. A part of the results already was in press (Biol.Pharm.Bull.,2005), and other was submitted and then be under examination. Less