缺血后处理通过上调磷酸化丝氨酸/苏氨酸蛋白激酶阻断细胞色素C释放保护缺血性神经元损伤

doi:10.3969/j.issn.0529-1356.2013.02.002

解剖学报 ›› 2013, Vol. 44 ›› Issue (2 ): 152-156.doi: 10.3969/j.issn.0529-1356.2013.02.002

缺血后处理通过上调磷酸化丝氨酸/苏氨酸蛋白激酶阻断细胞色素C释放保护缺血性神经元损伤

张立柱^1,2 周彩凤^1,3 涂静宜¹ 张茜¹ 朱莹¹ 王瑞敏^1*

1.河北联合大学医学实验研究中心神经生物学研究所，河北唐山 063000;2.唐山市乐亭县医院外科，河北唐山 063604; 3.沧州市中心医院科研处，河北沧州 061001

收稿日期:2012-03-23 修回日期:2012-07-12 出版日期:2013-04-06 发布日期:2013-04-06
通讯作者: 王瑞敏 E-mail:ruimin-wang@163.com
基金资助:
国家自然科学基金资助项目

Ischemic postconditioning prevents cytochrome C release through up-regulation of p-Akt and protects neurons against ischemia insult 

ZHANG Li-zhu ^1,2 ZHOU Cai-feng ^1,3 TU Jing-yi¹ ZHANG Xi¹ ZHU Ying¹ WANG Rui-min^1*

1. Institute of Neurobio logy, Medical Research Center, Hebei United University, Hebei Tangshang 063000,China;2.Surgical Department, Leting Country Hospital, Hebei Tangshang 063604,China;3.Reserch Department of Cangzhou Central Hospital, Hebei Cangzhou 061001,China

Received:2012-03-23 Revised:2012-07-12 Online:2013-04-06 Published:2013-04-06

摘要/Abstract

摘要：

目的通过观察脑缺血再灌注大鼠海马CA1区神经元内丝氨酸/苏氨酸蛋白激酶B（Akt）及细胞色素C 的表达及定位情况，探讨延迟性脑缺血后处理神经保护作用及其可能的分子机制。方法制作SPF级成年雄性SD大鼠（40只）四动脉结扎全脑缺血模型。实验动物随机分为假手术组，缺血再灌注组，后处理组，抑制剂组及溶剂对照组。采用焦油紫染色技术观察大鼠海马CA1区神经元存活情况；Western blotting法检测磷酸化Akt及细胞色素C的表达及定位。结果延迟性的脑缺血后处理能够促进缺血再灌注大鼠海马CA1区神经元生存；促进磷酸化Akt水平升高；阻止线粒体内细胞色素C向胞质释放；脑室注射LY294002后，细胞色素C的释放减少，抑制延迟性后处理的神经元保护作用。结论延迟性脑缺血后处理通过诱导胞质内Akt的磷酸化，抑制线粒体内细胞色素C释放到胞质，阻止全脑缺血再灌注后大鼠海马CA1区神经元损伤。

关键词: 全脑缺血 , 延迟性缺血后处理 , 丝氨酸/苏氨酸蛋白激酶B , 细胞色素C, 免疫印迹法 , 大鼠

Abstract:

Objective The goal of this study is to elucidate the neuro-protective effect and the possible mechanism of delayed ischemic postconditioning through observing the level of p-Akt and cytochrome C (Cyt C) in cytoplasm or mitochondria following global cerebral ischemia. Methods 40 Adult male Sprague-Dawley rats were subjected to global cerebral ischemia by four-vessel occlusion and were randomly divided into five groups: sham group, ischemia-reperfusion group (I/R), delayed ischemic postconditioning group (Post C), Vehicle group (Post C+Vehicle) and LY294002 group (Post C+LY294002). Cresyl violet staining was used to observe the surviving neurons of hippocampal CA1 region following global cerebral ischemia and western blot analysis was used to detect the level of p-Akt and Cyt C in both cytosolic and mitochondrial fraction. Results Delayed ischemic postconditioning protected the hippocampal CA1 region neurons against ischemia/referfusion injury and significantly increased the level of p-Akt in cytoplasm compared with I/R groups. Delayed ischemic postconditioning markedly prevented the release of Cyt C from mitochondria to cytoplasm and LY294002, an inhibitor of Akt up-stream kinase, abolished the positive role of delayed ischemic postconditioning. Conclusion Delayed ischemic postconditioning induces the Akt activation and prevents the release of Cyt C from mitochondria to cytoplasm, thereby blocks the hippocampal CA1 region neurons injury following global cerebral ischemia.

Key words: Global cerebral ischemia , Delayed ischemic postconditioning , Akt, Cytochrome C , Western blotting , Rat

中图分类号:

R363.2

张立柱周彩凤涂静宜张茜朱莹王瑞敏1* . 缺血后处理通过上调磷酸化丝氨酸/苏氨酸蛋白激酶阻断细胞色素C释放保护缺血性神经元损伤[J]. 解剖学报, 2013, 44(2 ): 152-156.

ZHANG Li-zhu ZHOU Cai-feng TU Jing-yi ZHANG Xi ZHU Ying WANG Rui-min* . Ischemic postconditioning prevents cytochrome C release through up-regulation of p-Akt and protects neurons against ischemia insult [J]. AAS, 2013, 44(2 ): 152-156.

参考文献

［1］ Tsang A, Hausenloy DJ, Mocanu MM, et al. Postconditioning: a form of “modified reperfusion” protects the myocardium by activating the phosphatidylinositol 3-kinase-Akt pathway［J］. Circ Res, 2004, 95(3):230-232.

［2］ Pignataro G, Meller R, Inoue K, et al. In vivoand in vitro characterization of a novel neuroprotective strategy for stroke: ischemic Post C［J］. J Cerebral Blood Flow Metab, 2008,28(2):232-241.

［3］ Rehni AK, Singh N. Role of phosphoinositide 3-kinase in ischemic Post C-induced attenuation of cerebral ischemia-evoked behavioral deficits in mice ［J］. Pharmacol Rep, 2007,59(2):192-198.

［4］ Zhao H. The protective effects of ischemic postconditioning against stroke: from rapid to delayed and remote postconditioning［J］. Open Drug Discov J, 2011,5:138-147.

［5］ Christophe M, Nicolas S. Mitochondria: a target for neuroprotective interventions in cerebral ischemiareperfusion［J］. Curr Pharm Des， 2006,12(6):739-757.

［6］ Danielisova V, Gottlieb M, Nemethova M, et al. Bradykinin postconditioning protects pyramidal CA1 neurons against delayed neuronal death in rat hippocampus［J］. Cell Mol Neurobiol, 2009,29(6-7):871-878.

［7］ Yang LC, Zhang QG, Zhou CF, et al. Extranuclear estrogen receptors mediate the neuroprotective effects of estrogen in the rat hippocampus［J］. PLoS One, 2010, 5(5): e9851.

［8］ Zhao ZQ, Vinten-Johansen J. Postconditioning: reduction of

Reperfusion-induced injury［J］. Cardiovasc Res, 2006, 70(2): 200-211.

［9］ Kim AH, Klarsigma G, Sun X, el a1. Akt phosphorylates and negatively regulates apoptosis signal-regaling kinase 1［J］. Mol Cell Biol, 2001, 21(3): 893-901

［10］ Gao X, Zhang H, Takahashi T, et al. The Akt signaling pathway contributes to postconditioning’s protection against stroke; the protection is associated with the MAPK and PKC pathways［J］. J Neurochem, 2008, 105(3):943-95

[11] Galluzzi L, Morselli E, Kepp O, et al. Targeting postmitochondrial effectors of apoptosis for neuroprotection ［J］. Biochim Biophys Acta, 2009,1787(5): 402-413.

［12］ Cao G, Luo Y, Nagayama T, et al. Cloning and characterization of rat Caspase-9: implications for a role in mediating Caspase-3 activation and hippocampal cell death after transient cerebral ischemia ［J］. J Cereb Blood Flow Metab, 2002, 22(5): 534-546.

［13］ Green DR, Reed JC. Mitochondria and apoptosis［J］. Science, 1998, 281(5381):1309-1312.

［14］ SugawaraT, Fujimura M, Morita-Fujimura Y, et al. Mitochondrial release of cytochrome C correponds to selective vulnerability of hippocampal CA1 neurons in rats after transient globe cerebral ischemia［J］. J Neurosci, 1999, 19(22): Rc39.

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缺血后处理通过上调磷酸化丝氨酸/苏氨酸蛋白激酶阻断细胞色素C释放保护缺血性神经元损伤

Ischemic postconditioning prevents cytochrome C release through up-regulation of p-Akt and protects neurons against ischemia insult 

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