苯丙氨酸及其衍生物对未成熟神经元胞质游离钙的影响_医学论文.docVIP

苯丙氨酸及其衍生物对未成熟神经元胞质游离钙的影响_医学论文 苯丙氨酸及其衍生物对未成熟神经元胞质游离钙的影响_医学论文 作者：张惠文，袁小兵，张拥军，顾学范 【关键词】 苯丙酮尿症 　　【Abstract】 AIM: To gain more insight into the mechanism of neuronal injury in phenylketonuria patients and to investigate the effect of phenylalanine on intracellular free calcium concentration (［Ca］i) of embryonic rat cortical neurons. METHODS: Calcium imaging was used to observe the influence of Lphenylalanine and its derivatives on ［Ca］i. RESULTS: High concentration of Lphenylalanine (Phe, 10 mmol/L) and DLβphenyllactic acid （PL,10 mmol/L）decreased ［Ca］i, while phenylacetic acid (PAA, 10 mmol/L) decreased, then increased, and again decreased ［Ca］i. With the normal extracellular balance solution replaced by calciumfree and natriumfree balance solution, PL did not change its effect on intracellular free calcium. With Thapsigargin (100 nmol/L) added to the normal extracellular solution to inhibit the sarcoplasmic/endoplasmic reticulum Ca2+ATPase（SERCA）, PAA decreased ［Ca］i, and no subsequent increase of ［Ca］i was observed. With the same concentration, PL and PAA had more significant effects on immature neurons. CONCLUSION: Combining the result of gene chips, Lphenylalanine and its derivatives can activate plasma membrane Ca2+ATPase and accelerate the extrusion of intracellular free calcium from neurons, thus resulting in the decrease of ［Ca］i. 　　【Keywords】 phenylketonuria calcium imaging phenylalanine 　　【摘要】 目的： 探讨苯丙酮尿症患者脑损伤的病理生理机制，分析苯丙氨酸和胞质游离钙有无关系. 方法： 用钙成像技术检测氨基酸及其衍生物对胚鼠皮层未成熟神经元胞质游离钙浓度的影响. 结果： 高浓度苯丙氨酸降低胞质游离钙，苯乙酸使胞质内钙浓度降低后升高最后又降低，苯乳酸使胞质内钙降低更明显. 使用无钙离子无钠离子的细胞外液，苯乳酸降低胞质内钙的作用无明显影响；使用Thapsigargin阻断内质网上钙泵，苯乙酸虽仍能降低胞质内钙，但没有降低后又升高的现象. 当苯丙氨酸、苯乙酸和苯乳酸相同浓度时，苯乙酸、苯乳酸较苯丙氨酸的作用更强. 结论： 结合我们以前的观察结果，苯丙氨酸可能激活plasma membrane Ca2+ATPase，促进神经元胞质内钙外排，进而导致神经元胞质内钙浓度降低. 　　【关键词】 苯丙酮尿症；钙成像；苯丙氨酸 　　0引言 　　苯丙酮尿症是遗传代谢性疾病，由于苯丙氨酸羟化酶基因突变导致的酶活性缺失或者降低，苯丙氨酸不能转化为酪氨酸，体内苯丙氨酸及其衍生物苯乙酸、苯丙酮酸、苯乳酸等贮积. 苯丙酮尿症的研究历史较长，生化研究显示高浓度的苯丙氨酸抑制Na+, K+ATP酶、肌酐激酶活性，电生理研究显示高浓度的苯丙氨酸抑制海马神经元N型钙通道开放，抑制NMDA受体激活，抑制突触前膜谷氨酸释放，抑制突触后谷氨酸能兴奋性电位［