白藜芦醇预处理对大鼠皮质神经元氧糖剥夺/再复氧损伤后神经元突起生长的影响

doi:10.16098/j.issn.0529-1356.2017.01.001

摘要/Abstract

摘要：

目的探讨白藜芦醇预处理对大鼠皮质神经元氧糖剥夺/再复氧损伤后神经元突起生长的影响。方法体外皮质神经元氧糖剥夺150min，复氧培养24 h。实验分为正常组、对照组和5μmol/L白藜芦醇预处理组。免疫荧光法鉴定神经元，细胞计数试剂盒8（CCK-8）法测定细胞活力，TUNEL法检测细胞凋亡，免疫荧光法和Western blotting检测微管相关蛋白2（MAP-2）和生长相关蛋白43（GAP-43）的表达,并计数神经元突起的长度和数目。结果培养细胞均高表达神经元特异性标记物MAP-2。白藜芦醇组细胞活力较对照组明显增加（0.551±0.009 比 0.436±0.013, P<0.01），而凋亡则明显降低（18.3%±1.3% 比35.3%±1.9%, P<0.01），上调MAP-2(0.790±0.102 比 0.462±0.063, P<0.01)和GAP-43(0.768±0.084 比 0.424±0.065,P<0.01)蛋白的表达，增加神经元突起的长度(89.510±6.939 比 61.538±9.14,P<0.01)和数目(6.347±1.002 比 3.040±0.608,P<0.01）。结论白藜芦醇预处理能减轻氧糖剥夺/再复氧对神经元的损伤，促进神经元突起的生长。

关键词: 白藜芦醇, 神经保护, 神经突起, 氧糖剥夺/再复氧, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the effects of resveratrol pretreatment on neurite growth of rat primary cortical neurons after oxygen-glucose deprivation/reperfusion (OGD/R) injury in vitro. Methods Primary cortical neurons were cultured under oxygen and glucose deprivation for 150 minutes and reoxygenation for 24 hours. The study had the normal, control and 5μmol /L resveratrol pretreatment groups. Neurons were identified with immunofluorescence. Cell viability was detected with cell counting kit-8(CCK-8) assay. Cell apoptosis was detected with TUNEL assay. Immunofluorescence and Western blotting measured the expressions of microtubule-associated protein 2（MAP-2）and growth associated protein 43（GAP-43）, and the length and number of neurites were counted.
Results Cells had high expression of neuronal specific marker MAP-2. Compared with the control group, resveratrol treatment significantly enhanced the neurons viability (0.551±0.009 vs 0.436±0.013,P<0.01), decreased the numbers of apoptosis (18.3%±1.3% vs 35.3%±1.9%,P<0.01), upregulated the expressions of MAP-2 (0.790±0.102 vs 0.462±0.063,P<0.01) and GAP-43 (0.768±0.084 vs 0.424±0.065,P<0.01) proteins, increased the length (89.510±6.939 vs 61.538±9.14,P<0.01) and numbers (6.347±1.002 vs 3.040±0.608,P<0.01) of neurites. Conclusion Resveratrol pretreatment can reduce injury and promote neurite growth of cultured neurons after OGD/R.

Key words: Resveratrol, Neuroprotection, Neurite, Oxygen-glucose deprivation/reoxygenation, Western blotting, Rat

唐凡人余萍萍王莉郭霜杨琴. 白藜芦醇预处理对大鼠皮质神经元氧糖剥夺/再复氧损伤后神经元突起生长的影响[J]. 解剖学报, 2017, 48(1): 1-6.

TANG Fan-ren YU Ping-ping WANG Li GUO Shuang YANG Qin. Effects of resveratrol pretreatment on neurite growth of primary cortical neurons after oxygen-glucose deprivation/reperfusion injury in rats[J]. Acta Anatomica Sinica, 2017, 48(1): 1-6.

参考文献

［1］Lopez M, Dempsey R, Vemuganti R. Resveratrol neuroprotection in stroke and traumatic CNS injury［J］.Neurochem Int, 2015, 89:75-82.

［2］Cheng W, Shen ChB,Wang L,et al. Effect of resveratrol pretreatment on proliferation of cortical neural stem cells after oxygen-glucose deprivation /reperfusion injury in rat［J］.Chinese Pharmacological Bulletin, 2015, 31(1):113-118.(in Chinese)

成薇,沈长波,王莉,等.白藜芦醇预处理对氧糖剥夺/再复氧损伤大鼠皮质神经干细胞增殖的影响［J］.中国药理学通报,2015,31(1):113-118.

［3］Cheng W,Yu P, Wang L,et al. Sonic hedgehog signaling mediates resveratrol to increase proliferation of neural stem cells after oxygen-glucose deprivation/reoxygenation injury in vitro［J］. Cell Physiol Biochem, 2015, 35(5):2019-2032.

［4］Zhang LL, Huang JG, Shen ChB,et al. Effect of resveratrol on rat primary cortical neurons during oxygen and glucose deprivation /reperfusion injury［J］.Chinese Traditional Patent Medicine,2014,36(5):897-903.(in Chinese)

张黎黎,黄家贵,沈长波，等. 白藜芦醇预处理对缺糖缺氧再灌注大鼠原代皮质神经元的保护作用［J］.中成药,2014,36(5):897-903.

［5］Yu P,Wang L,Tang F,et al. Resveratrol pretreatment decreases ischemic injury and improves neurological function via sonic hedgehog signaling after stroke in rats. ［J］. Mol Neurobiol, doi:10-1007/s12035-015-9639-7.

［6］Yu PP,Wang L,Tang FR,et al. Effect of resveratrol on activation of astrocyte after cerebral ischemia /reperfusion injury in rats［J］. Chinese Pharmacological Bulletin, 2015, 31(9):1228-1233.(in Chinese)

余萍萍,王莉,唐凡人,等. 白藜芦醇对大鼠脑缺血/再灌注损伤后星形胶质细胞活化的影响［J］.中国药理学通报,2015,31(9):1228-1233.

［7］Dasgupta B, Milbrandt J. Resveratrol stimulates AMP kinase activity in neurons［J］. Proc Natl Acad Sci USA, 2007, 104(17):7217-7222.

［8］Bora-Tatar G, Erdem-Yurter H. Investigations of curcumin and resveratrol on neurite outgrowth: perspectives on spinal muscular atrophy［J］. Biomed Res Int, 2014, 2014:709108. ［9］Rong Z,Pan R,Xu Y,et al. Hesperidin pretreatment protects hypoxia-ischemic brain injury in neonatal rat［J］. Neuroscience, 2013, 255:292-299.

［10］Li Z, Fang F, Wang Y. Resveratrol protects CA1 neurons against focal cerebral ischemic reperfusion-induced damage via the ERK-CREB signaling pathway in rats［J］.Pharmacol Biochem Behav, 2016, 146-147:21-27.

［11］Huang JG,Zhang LL,Shen ChB,et al. Effects of resveratrol on oxygen and glucose deprivation /reoxygenation injury of rat primary cortical neurons at different time windows［J］. Chinese Pharmacological Bulletin,2013,29(9):1230-1234.(in Chinese)

黄家贵,张黎黎,沈长波,等. 白藜芦醇在缺糖缺氧/再灌注损伤不同时间窗对大鼠皮质神经元的保护作用［J］. 中国药理学通报,2013,29(9):1230-1234.

［12］Wang Q,Yang L,Wang Y. Enhanced differentiation of neural stem cells to neurons and promotion of neurite outgrowth by oxygen-glucose deprivation［J］.Int J Dev Neurosci, 2015, 43:50-57.

［13］Shen Y,Karina M. GAP-43 dependency defines distinct effects of netrin-1 on cortical and spinal neurite outgrowth and directional guidance［J］. Dev Neurosci, 2013, 31(1):11-20.

［14］Tao T, Feng J, Xu G. Minocycline promotes neurite outgrowth of PC12 cells exposed to oxygen-glucose deprivation and reoxygenation through regulation of MLCP_MLC signaling pathways［J］. Cell Mol Neurobiol, doi:10.1007/s10571-016-0374-z.

［15］Manczak M, Mao P, Calkins MJ, et al. Mitochondria-targeted antioxidants protect against amyloid-beta toxicity in Alzheimer’s disease neurons［J］. J Alzheimers Dis, 2010, 20 (Suppl 2):S609-631.

［16］Fan L, Simard LR. Survival motor neuron (SMN) protein: role in neurite outgrowth and neuromuscular maturation during neuronal differentiation and development［J］. Hum Mol Genet, 2002, 11(14):1605-1614.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	谢惠兰方芳林毅. Chir99021调控Wnt/β-catenin 信号通路抑制大鼠牙髓干细胞成骨诱导分化的机制[J]. 解剖学报, 2024, 55(1): 67-72.
[3]	魏茜茜李超红赵晨露唐家萍刘玉珍. 大鼠颈上神经节中Ⅰ组代谢型谷氨酸受体表达及慢性间歇性低氧对其的影响 [J]. 解剖学报, 2024, 55(1): 3-9.
[4]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[5]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[6]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[7]	吕海燕沈西雅赵富生朱梅. 松茸多糖对 1-甲基-4-苯基-吡啶离子诱导 PC12 细胞损伤的影响 [J]. 解剖学报, 2024, 55(1): 49-54.
[8]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[9]	苏芃王兴仪梁景岩熊天庆. 一种低密度及高纯度胎鼠原代海马神经元培养方法的优化及鉴定[J]. 解剖学报, 2024, 55(1): 113-119.
[10]	郝永明郭磊周程辉裴汉军李莉赵哲 . 改良腹主动脉缩窄法建立心肌肥厚模型[J]. 解剖学报, 2024, 55(1): 120-124.
[11]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[12]	刘丽平朱梦妮徐慧 . 白细胞介素6对脂多糖诱导所致类子痫前期大鼠有核红细胞的影响 [J]. 解剖学报, 2023, 54(6): 722-729.
[13]	赵卫明李晓余国营王改平常翠芳 . 丝氨酸肽酶抑制因子Kazal型1对肝癌细胞增殖的影响及其机制 [J]. 解剖学报, 2023, 54(6): 695-702.
[14]	王文娟丁雨桐苏昊任捷孙艳荣王涵菲陆嘉莉张林倩白钰秦丽华. 低雌激素状态下大鼠海马及纹状体Nogo-A的表达变化[J]. 解剖学报, 2023, 54(6): 620-627.
[15]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.