苯丙酮尿症大鼠海马神经元损伤及其与BDNF/TrkB通路的关系-东南大学学报医学版

苯丙酮尿症大鼠海马神经元损伤及其与BDNF/TrkB通路的关系

单位：郑州大学第三附属医院新生儿筛查中心, 河南郑州 450000

分类号：R-33;R742.8

出版年·卷·期（页码）：2019·38·第三期（461-466）

摘要：

目的：观察苯丙酮尿症大鼠海马神经元损伤及其与脑源性生长因子（BDNF）/酪氨酸激酶受体B（TrkB）通路的关系。方法：将24只大鼠根据随机数字表法分为对照组和苯丙酮尿症组各12只。用10% FeCl₃液显色进行尿液实验，采用高效液相色谱-荧光检测法测定大鼠大脑皮层及血清中酪氨酸和苯丙氨酸水平，HE染色观察海马CA1区病理变化，采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定（TUNEL）法测定神经元凋亡情况，采用蛋白质印迹法测定海马含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase-3）、B淋巴细胞瘤-2基因（Bcl-2）蛋白和BDNF、TrkB、磷酸化细胞外调节蛋白激酶（p-ERK）、磷酸化丝氨酸/苏氨酸激酶（P-Akt）蛋白含量。结果：苯丙酮尿症组尿液显色为强阳性；大脑皮层和血清苯丙氨酸水平明显高于对照组（P<0.05），大脑皮层和血清酪氨酸水平低于对照组（P<0.05）；海马CA1区组织结构紊乱，细胞结构模糊、排列紊乱，见神经元细胞变性、坏死；海马CA1区细胞凋亡数高于对照组（P<0.05）；海马Caspase-3、Bcl-2、BDNF、TrkB、p-ERK、P-Akt蛋白含量高于对照组（P<0.05）。结论：苯丙酮尿症大鼠海马神经元存在损伤，其机制可能与BDNF/TrkB通路有关。

Objective:To observe the relationship between hippocampal neuron injury in rats with phenylketonuria and the brain-derived growth factor(BDNF)/tyrosine kinase receptor B(TrkB) pathway, and to explore the possible mechanism of hippocampal neuron injury. Methods:Twenty-four rats were divided into control group and phenylketonuria group by random number method with 12 rats in each group. Urine test was conducted with 10%FeCl₃ liquid chromogenic analysis. Tyrosine and phenylalanine levels in cerebral cortex and serum were determined by HPLC-fluorescence detection. Pathological changes in the CA1 region of the hippocampus were observed by HE staining. Levels of cysteine-containing aspartate proteolytic enzyme 3(Caspase-3), b-lymphocytoma-2(bcl-2), BDNF, TrkB, phosphorylated extracellular regulatory protein kinase(p-erk), and phosphorylated serine/threonine kinase(P-Akt) proteins in hippocampus were determined by Western blot. Results:The urine of rats in phenylketonuria group was strongly positive. The levels of phenylalanine in the cerebral cortex and serum in the phenylketonuria group were significantly higher than those of the control group(P<0.05), while the levels of phenylalanine in the cerebral cortex and serum in the phenylketonuria group were significantly lower than those of the control group(P<0.05). In phenylketonuria group, the tissue structure in CA1 area of the hippocampus was disordered, and the cell structure was fuzzy and disordered. The degeneration and necrosis of neurons were observed. The number of apoptic cells in the CA1 region of hippocampus in phenylketonuria group was higher than that of the control group(P<0.05). The levels of Caspase-3, Bcl-2, BDNF, TrkB, p-erk and p-akt protein in hippocampus of the phenylketonuria group were higher than those of the control group(P<0.05). Conclusion:The mechanism of hippocampal neurons injury in phenylketonuria rats may be related to the BDNF/TrkB pathway.

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