苯丙氨酸及其代谢物对P19细胞神经分化的影响

探讨苯丙氨酸及其代谢物对P19细胞分化的影响及机制。实验各组从P19细胞神经分化诱导期开始分别加入苯丙氨酸、苯乙酸和苯乳酸 ,观察神经分化期细胞形态 ,MTT法检测神经分化期细胞存活率和LDH漏出量的变化 ,并通过流式细胞仪检测诱导后细胞周期有无变化。结果显示 :苯丙氨酸及其代谢物可使P19神经元肿胀、崩解 ,神经突起纤细且突起数少 ,可降低P19细胞存活率 ,但对LDH漏出量并无影响 ,对诱导后的P19细胞周期也无影响。以上结果提示 ,苯丙氨酸及其代谢物可干扰P19细胞的神经诱导过程 ,母源性苯丙酮尿症中枢神经系统损伤可能与其在脑发育早期阶段神经诱导及分化过程中受到高苯丙氨酸及其代谢物影响有关。

Effects of phenylalanine and its metabolites on the neuronal differentiation of P19 cell

The study was undertaken to determine the effect and mechanism of phenylalanine (PHE)and its metabolites on the neuronal differentiation of P19 embrional carcinomal cell(P19 cell). PHE or phenylacetic acid (PAA) or phenyllactic acid (PLA) was used to treat P19 cell from induction phase till the end of the experiment. After being plated for neuronal differentiation, cell morphology was observed under light microscope. P19 cell viability was determined by mitochondrial MTT reduction and cellular LDH release. Cell cycle analysis was performed by using flow cytometry. The result indicated that P19 neurons exposed to PHE and its metabolites showed various changes such as cell swelling, collapsing, slender neurofibrils, decrease in the number of neurofibrils, etc. PHE, PAA, PLA remarkably decreased the viability of P19 cells without altering cellular LDH release. Flow cytometry revealed that PHE, PAA, PLA had no effect on the cell cycle of the P19 cell after induction phase. Our data showed PHE or its metabolites could disturb the neuronal induction of P19 cell and suggested that PHE or its metabolites might be involved in the central nervous systerm dysfunction associated with maternal PKU. [