Oxygen-Glucose-Deprivation / Reoxygenation-Induced Autophagic Cell Death Depends on JNK-Mediated Phosphorylation of Bcl-2
Oxygen-Glucose-Deprivation / Reoxygenation-Induced Autophagic Cell Death Depends on JNK-Mediated Phosphorylation of Bcl-2
复制标题
缺氧/葡萄糖剥夺/复氧诱导的自噬细胞死亡取决于 JNK 介导的 Bcl-2 磷酸化
DOI：
10.1159/000443057
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发表时间：
2016-01-01
影响因子：
--
通讯作者：
Yin, Guoyong
中科院分区：
文献类型：
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作者：
Fan, Jin;Liu, Yuwen;Yin, Guoyong
背景/目的：本研究旨在探讨自噬在大鼠神经元缺氧缺糖/复氧(OGD/R)损伤中的作用。方法和结果：分离SD大鼠大脑皮质神经元，免疫荧光法鉴定。将大鼠大脑皮质神经元随机分为4组：对照组(I)、实验组(II)、JNK抑制剂预处理组(III)和JNK抑制剂+OGD/R组(IV)。IV组复氧6h和12h后神经细胞存活率显著降低(P<；0.05)。电子显微镜下可见神经元内有大量自噬空泡和自溶酶体形成，6h自噬空泡数量短暂减少，12h出现新的自噬通量和大量空泡。IV组复氧0.5h和2h出现大量自噬空泡，随着复氧时间的延长，自噬空泡逐渐减少。复氧6h前，II组和IV组的LC3II蛋白表达差异无统计学意义，总体呈上升-下降模式。随着复氧时间的延长，Ⅱ组的磷酸化JNK、Bcl2和Beclin-1的表达逐渐升高，而Ⅳ组的表达在复氧12h时逐渐升高(P<0.05)。此外，在II组中观察到Bcl2/Beclin-1复合体的渐进性解离，而JNK抑制剂抑制了这种解离。结论：JNK/Bcl2/Beclin-1信号通路的调控可能是OGD/R诱导神经元自噬细胞死亡的机制之一。版权所有(C)2016 S.Karger AG，巴塞尔
Background/Aims: The purpose of this study was to investigate the role of autophagy in oxygen-glucose-deprivation/reoxygenation (OGD/R) injury in rat neurons. Methods and results: Cortical neurons were isolated from Sprague-Dawley rats and identified by immunofluorescence. The cortical neurons were randomly assigned to one of four groups: control group (I), experimental group (OGD/R group, II), JNK inhibitor pretreatment group (III) and JNK inhibitor pretreatment + OGD/R group (IV). Neuronal cell viability significantly decreased after 6h and 12h of reoxygenation in Group IV (P < 0.05). Electron microscopy showed the presence of many autophagic vacuoles and the formation of autolysosomes in the neurons; the number of autophagic vacuoles decreased transiently at 6h, while a new autophagic flux and a large number of empty autophagic vacuoles were observed at 12h. In Group IV, a large number of autophagic vacuoles were present at 0.5h and 2h of reoxygenation, which gradually decreased with increasing reoxygenation time. No significant differences in the expression of the LC3II protein were detected between the Group II and IV prior to 6h of reoxygenation, and LC3II expression showed an overall rise-decline pattern. However, LC3II protein expression increased in Group II at 12h of reoxygenation, whereas a continuous decline was observed in Group IV. The levels of phosphorylated JNK and Bcl-2 and the expression of Beclin-1 increased gradually as the reoxygenation time going in Group II, whereas they increased at 12h of reoxygenation in Group IV (P < 0.05). In addition, progressive dissociation of the Bcl-2/Beclin-1 complex was observed in the Group II, while JNK inhibitor suppressed this dissociation. Conclusion: The regulation of the JNK/Bcl-2/Beclin-1 signaling pathway may be one of the mechanisms underlying the OGD/R-induced autophagic cell death of neurons. Copyright (C) 2016 S. Karger AG, Basel