黄蜀葵花总黄酮对海马神经元缺氧复氧损伤的保护作用

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (4): 367-372.

黄蜀葵花总黄酮对海马神经元缺氧复氧损伤的保护作用

文继月，陈志武

安徽医科大学，基础医学院药理学教研室，合肥 230032，安徽

收稿日期:2016-12-05 修回日期:2017-01-11 出版日期:2017-04-26 发布日期:2017-04-26
通讯作者: 陈志武，男，博士，教授，研究方向：心脑血管药理学。 Tel：13515669237 E-mail：chpharmzw@163.com
作者简介:文继月，女，博士，讲师，研究方向：心脑血管药理学。 Tel：18256510760 E-mail：wenjiyue139@aliyun.com.cn
基金资助:
国家自然科学基金（81173596）

Protective effect of total flavones of abelmoschl manihot on hippocampal neurons subjected to anoxia-reoxygenation injury

WEN Jiyue, CHEN Zhiwu

Department of Pharmacology, Anhui Medical University, Hefei 230032, Anhui, China

Received:2016-12-05 Revised:2017-01-11 Online:2017-04-26 Published:2017-04-26

摘要/Abstract

摘要：

目的: 研究黄蜀葵花总黄酮（total flavones of abelmoschl manihot,TFA）对海马神经元缺氧复氧损伤的保护作用以及可能的内皮衍生超级化因子（endothelium-dependent hyperpolarizing factor,EDHF）相关机制。方法: 采用原代培养的海马神经元建立缺氧4 h/复氧24 h损伤模型，预先在细胞培养上清内加入大鼠脑基底环动脉（basilar artery,BA）环，分别给予乙酰胆碱（acetylcholine,Ach）联合一氧化氮（nitric oxide,NO）合酶抑制剂Nω-nitro-L-arginine-methyl-ester（L-NAME)和前列环素（prostacyclin,PGI2）抑制剂indomethacin（Indo）；TFA联合L-NAME和Indo预处理。检测缺氧/复氧损伤后海马神经元存活率、培养上清液中乳酸脱氢酶（lactate dehydrogenase,LDH）活力的变化；另外，用激光共聚焦法检测细胞内游离Ca²⁺浓度。结果: 与单独应用Ach或者TFA预处理比较，Ach或者TFA联合脑基底动脉环均能明显改善缺氧/复氧后海马神经元存活率的降低，培养上清中LDH活力降低。此外，TFA联合脑基底动脉环抑制海马神经元细胞内游离Ca²⁺的增高，L-NAME及Indo对此结果并无显著影响。结论: TFA对缺氧/复氧致海马神经元损伤有明显的保护作用，其机制可能与促进EDHF释放，抑制细胞内Ca²⁺超载有关。

关键词: 黄蜀葵花总黄酮, 内皮衍生超级化因子, 缺氧/复氧损伤, 乳酸脱氢酶, 钙超载

Abstract:

AIM: To study the protective effect of total flavones of abelmoschl manihot (TFA) on hippocampal neurons subjected to the injury of anoxia/re-oxygenation (A/R) and its endothelium-dependent hyperpolarizing factor (EDHF) mechanism. METHODS: The injury of primarily cultured hippocampal neurons plus with cerebral basilar artery ring was induced by hypoxia for 4 hours and re-oxygenation for 24 hours, pretreated with TFA, acetylcholine (Ach),etc. Cell viability and the lactate dehydrogenase (LDH) activity in supernatant were served as the cell damage index. The level of free calcium fluorescence intensity in the neurons was detected by laser scanning confocal microscope. RESULTS: Pretreated with TFA and cerebral artery could increase the cell viability, reduce the activity of LDH in culture medium significantly, pretreated with Ach had a similiar effect. The N -nitro-L-arginine-methyl-ester (L-NAME) and indomethacin (Indo) didn't influence the effect of Ach or TFA. Moreover, pretreated with TFA plus with cerebral artery lowered the calcium concentration in neuron (compared with model group). CONCLUSION:TFA has remarkably neuroprotective effect induced by A/R, the mechanism is partially concerned with the calcium overload inhibited by EDHF released from cerebral basilar artery ring.

Key words: TFA, EDHF, A/R injury, LDH, calcium concentration

中图分类号:

R965.2

文继月，陈志武. 黄蜀葵花总黄酮对海马神经元缺氧复氧损伤的保护作用[J]. 中国临床药理学与治疗学, 2017, 22(4): 367-372.

WEN Jiyue, CHEN Zhiwu. Protective effect of total flavones of abelmoschl manihot on hippocampal neurons subjected to anoxia-reoxygenation injury[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(4): 367-372.

参考文献

［1］陈国成，潘林平，陈巧明，等. 中医特色卒中单元对缺血性脑卒中患者生活质量的影响［J］.长春中医药大学学报,2015, 31（6）:1182-1185.
［2］Damkjaer M, Nielsen G, Bodendiek S, et al. Pharmacological activation of KCa3.1/KCa2.3 channels produces endothelial hyperpolarization and lowers blood pressure in conscious dogs［J］. Br J Pharmacol, 2012, 165(1): 223-234.
［3］Bos EM, Wang R, Snijder PM, et al. Cystathionine γ-lyase protects against renal is chemia/reperfusion by modulating oxidative stress［J］. J Am Soc Nephrol, 2013, 24(5): 759-770.
［4］吴剑，陈志武. 乙酰胆碱介导的内皮超极化因子对海马神经元缺氧/再给氧损伤的保护作用［J］. 中国药理学通报, 2007, 23(12): 1624-1629.
［5］Han J, Chen ZW, He GW. Acetylcholine and sodium hydrosulfide-induced endothelium- dependent relaxation and hyperpolarization in cerebral vessels of global cerebral ischemia- reperfusion rat［J］. J Pharmacol Sci, 2013, 121(4): 318-326
［6］高杉，范丽，董六一, 等. 黄蜀葵花总黄酮对MCAO大鼠脑细胞凋亡的影响［J］. 中国药理学通报, 2003, 19(6): 704-707.
［7］范丽，袁丽萍，陈志武, 等. 黄蜀葵花总黄酮药理性预适应对家兔心肌缺血再灌注损伤的保护作用［J］. 中国药学杂志, 2005, 40(11): 836-839.
［8］文继月，陈志武. 黄蜀葵花总黄酮预处理对大鼠脑缺血再灌注性损伤的保护作用［J］. 安徽医科大学学报, 2006, 41(6): 667-669.
［9］Bonde C, Noraberg J, Noer H, et al. Ionotropic glutamate receptors and gltamalte transporters are involved in necrotic cell death induced by oxygen-glucose deprivation of hippocampal slice cultures［J］. Neuroscience, 2005, 136(3): 779-794.
［10］张传林，王蕊，马克涛, 等. 大黄素增强SD大鼠脑极低动脉平滑肌细胞BKca电流［J］. 药理学通报, 2013, 29(9): 1244-1247.
［11］刘立，王友联，张枢, 等. 环孢霉素A对局部脑缺血再灌注损伤的改善作用［J］. 中风与神经疾病, 2002, 19(2): 106-107.
［12］Feletou M, Vanhoutte PM. Endothelium-dependent hyperpolarization of vascular smooth muscle cells［J］. Acta Pharm Sin, 2000, 21(1): 1-18.
［13］Yang Q, Yim AP, He GW. The significance of endotheliumderived hyperpolarizing factor in the human circulation［J］. Curr Vasc Pharmacol, 2007, 5(1): 85-92.
［14］Zhang RZ, Yang Q, Yim AP, et al. Role of NO and EDHF-mediated endothelial function in the porcine pulmonary circulation: comparison between pulmonary artery and vein［J］. Vascul Pharmacol, 2006, 44(3): 183-191.
［15］Dong YY, Wu M, Yim AP, et al. Hypoxia-reoxygenation, St. Thomas cardioplegic solution, and nicorandil on endotheliumderived hyperpolarizing factor in coronary microarteries［J］. Ann Thorac Surg, 2005, 80(5): 1803-1811.
［16］Takel N, Endo Y. Ca ²⁺ ionophre-induced apoptosis on cultured embryonic rat cortical neurons［J］. Brain Res, 1994, 652(1): 65-70.

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