黄芪注射液拮抗大鼠脑缺血再灌注损伤的保护作用和机制

doi:10.3969/j.issn.0529-1356.2014.03.003

摘要/Abstract

摘要：

目的探讨黄芪注射液对大鼠脑缺血再灌注损伤的神经保护作用和机制。方法成年健康雄性Wistar大鼠70只，应用线栓法经左侧颈外-颈内动脉插线建立大脑中动脉阻塞再灌注模型，经腹腔注射1g/L黄芪注射液（3ml/kg）干预治疗。Longa法评价大鼠神经功能缺损程度，氯化三苯基四氮唑（TTC）染色观察脑梗死体积，苏木素-伊红染色观察顶叶皮质区神经元形态结构变化，透射电子显微镜观察神经细胞的超微结构，流式细胞术检测细胞凋亡，Western blotting检测c-Jun氨基末端激酶3（JNK3）蛋白表达，RT-PCR检测JNK3 mRNA表达。结果经黄芪注射液治疗后，大鼠顶叶皮质区神经元JNK3 mRNA和JNK3蛋白表达较对照组显著减低，凋亡细胞数量显著减少，脑梗死体积显著缩小，神经元形态和超微结构以及动物神经行为功能显著改善。结论黄芪注射液可能通过下调神经元JNK3基因表达而抑制细胞凋亡，缩小脑梗死体积，改善动物的神经行为功能。

关键词: 脑缺血, 再灌注损伤, 黄芪注射液, c-Jun氨基末端激酶3, 凋, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the neuroprotective effect and mechanism of astragalus injection in cerebral ischemia reperfusion injury in rats. Methods A total of 70 healthy adult maleWistar rats were subjected to establish middle cerebral artery occlusion reperfusion models by inserting a monofilament thread from the external-internal carotid artery and treated by injecting 1g/L intraperitoneally astragalus injection (3ml/kg). The neurobehavioral function of rats was evaluated by Longa’s test and the cerebral infarction volume was calculated by tetrazolium chloride staining. The shape and ultrastructure of neurons in parietal cortex were observed by HE stain TEM. The early apoptotic ratio of neurons was detected by flow cytometry. The expressions of c-Jun N-terminal kinase 3 (JNK3) mRNA and protein were determined by RT-PCR and Western blotting, respectively. Results After treatment with astragalus injection, the expressions of JNK3 mRNA and protein reduced significantly, the number of neuronal apoptosis in parietal cortexminus, the cerebral infarction volume shrink, the neuronal shape and ultrastructure and animal neurobehavioral function were improved significantly than those in model group rats. Conclusion The results suggest that astragalus injection may inhibit neuronal apoptosis, reduce the infarction volume and improve the animal neurobehavioral function by down-regulated the expression of JNK3 gene following cerebral ischemia in rats.

Key words: Cerebral ischemia, Reperfusion injury, Astragalus injection, c-Jun N-terminal kinase 3, Apoptosis, Western blotting, Rat

王婷婷滕蕾* 黄惠李艳孙丽. 黄芪注射液拮抗大鼠脑缺血再灌注损伤的保护作用和机制[J]. 解剖学报, 2014, 45(3): 310-315.

WANG Ting-ting TENG Lei* HUANG Hui LI Yan SUN Li . Protective effect and mechanism of astragalus injection in cerebral ischemia reperfusion injury in rats[J]. AAS, 2014, 45(3): 310-315.

参考文献

［1］Blomgren K, Zhu C, Hallin U, et al. Mitochondria and ischemic reperfusion damage in the adult and in the developing brain［J］. Biochem Biophys Res Commun, 2003,304(3):551-559.
［2］Davis RJ. Signal transduction by the JNK group of MAP kinases ［J］. Cell, 2000, 103(2): 239-252.
［3］Ginet V, Puyal J, Magnin G, et al. Limited role of the c-Jun N-terminal kinase pathway in a neonatal rat model of cerebral hypoxia-ischemia［J］. J Neurochem, 2009, 108(3):552-562.
［4］Bogoyevitch MA, Kobe B. Uses for JNK: the many and varied substrates of the c-Jun N-terminal kinases［J］. Microbiol Mol Biol Rev, 2006, 70(4):1061-1095.
［5］Kuan CY, Wtfitmamh AJ, Yang DD, et al. A critical role of nalml-specific JNK3 for ischemic apoptosis［J］. Proe Nail Acad Sci USA, 2003, 100(25):15184-15189.
［6］Pirianov G, Brywe KG, Mallard C, et al. Deletion ofthe c-Jun N-ter-minal kinase 3 gene protects neonatal mice against cerebral hypoxic-ischemic injury［J］. J Cereb Blood Flow Metab, 2007, 27(5):1022-1032.
［7］O’Collins VE, Macleod MR, Donnan GA, et al. 1,026 experimental treatments in acute stroke［J］. Ann Neurol, 2006, 59(3):467-477. 
［8］Lijima T, Mishima T, Akagawa K, et al. Neuroprotective effect of propofol on necrosis and apoptosis following oxygen-glucose deprivation-relationship between mitochondrial membrane potential and mode of death［J］. Brain Res, 2006, 1099(1):25-32.
［9］Lai Zh, Yao CK, Cheng ShB, et al. Study of radix astragali on neuronal apoptosis in reperfusion local cerebral ischemia rats brains［J］. Journal of Emergency in Traditional Chinese Medicine, 2008, 17 (11):1565-1569.(in Chinese)
赖真, 姚灿坤, 程少冰, 等. 黄芪对脑缺血再灌注后的神经元凋亡的影响及机究［J］. 中国中医急症, 2008,17(11):1565-1569.
［10］Zhuo M, Chen HL, Cai NL, et al. A study on clinical effects and immunological mechanism of intravenous astragalus injection therapy in neonates with hypoxic ischemic encephalopathy［J］. Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care, 2008,15(1):13-15. (in Chinese)
卓名, 陈鸿莲, 蔡娜丽, 等. 黄芪注射液对新生儿缺氧缺血性脑病的临床与免疫机制研究［J］. 中国中西医结合急救杂志, 2008,15(1):13-15.
［11］Ye DQ, Gao WJ, Qian T, et al. Astragalus injection inhibits the expression of JNK3 after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats［J］. Chinese Pharmacological Bulletin, 2010, 26(1):77-82. (in Chinese)
叶冬青, 高维娟, 钱涛, 等. 黄芪注射液抑制缺氧缺糖/复氧复糖大鼠海马神经元JNK3的表达［J］. 中国药理学通报, 2010,26(1):77-82.
［12］Qu YZh, Li M, Gao GD. Protective effects of astragalus injection on blood-brain barrier and expressions of occludin and zonula occludens protein-1 after cerebral ischemia and reperfusion in rats［J］. Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care, 2011, 18(5):263-265. (in Chinese)
曲友直, 李敏, 高国栋. 黄芪注射液对大鼠脑缺血/再灌注后血脑屏障的保护作用及其机制研究［J］. 中国中西医结合急救杂志, 2011,18(5): 263-265.
［13］Longa EZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral artery occlusion without craniotomy in rats［J］. Stroke, 1989,20(1):84-91.
［14］Liu ShSh, Gao WJ, Qian T, et al. Effect of astragalus on expression of JNK3 in hippocampus of cerebral ischemia reperfusion rats［J］. Chinese Pharmacological Bulletin, 2012,28(5): 665-670. (in Chinese)
刘莎莎，高维娟，钱涛, 等. 黄芪注射液对脑缺血/再灌注大鼠海马组织JNK-3 表达的影响［J］. 中国药理学通报, 2012,28(5):665-670.
［15］Sun W, Gould TW, Newbem J, et al. Phosphorylation of c-Jun in avian and mammalian motoneurons in vivo during apoptosis: an early reversible event in the apoptotic casccade［J］. J Neuroscience, 2005,25(23): 5595-5603.
［16］Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases［J］. Science, 2002,298(5600):1911-1912.
［17］Davis RJ. Signal transduction by the JNK group of MAP kinases［J］. Cell, 2000, 103(2): 239-252.
［18］Nijboer CH, Heijnen CJ, Groenendaal F, et al. Alternate pathways preserve tumor necrosis factorαproduction after nuclear factorκB inhibition in neonatal cerebral hypoxia/ischemia［J］. Stroke, 2009,40(10):3362-3368.
［19］Guan QH, Pei DS, Liu XM, et al. Neuroprotection against ischemic brain injury by SP600125 via suppressing the extrinsic and intrinsic pathways of apoptosis［J］. Brain Res, 2006, 1092(1):36-46
［20］Ren F, Gong ShY, Zhi LM, et al. Effectiveness of astragalus for cerebral ischemic injury［J］. Chinese Journal of Clinical Rehabilitation, 2006, 10(3):149-151. (in Chinese)
任非, 龚淑英, 智丽敏, 等. 黄芪治疗缺血性脑损伤的有效性［J］. 中国临床康复, 2006, 10(3):149-151.
［21］Yan FX, Gao WJ, Qian T, et al. The effect of astragalus injection on the expression of Caspese-3 after hyposia/ hypoglycemia and re-oxygenation in hippocampal neurons of rats［J］. Chinese Pharmacological Bulletin, 2010, 26(7): 898-903. (in Chinese)
严凤霞, 高维娟, 钱涛, 等. 黄芪注射液对缺氧缺糖/复氧复糖大鼠海马神经元Caspase-3的表达影响［J］. 中国药理学通报, 2010, 26(7): 898-903.

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