目的 探讨丹灯通脑胶囊对脑缺血再灌注损伤大鼠的保护作用及可能作用机制,对丹灯通脑胶囊临床药效再评价。方法 采用线栓法复制大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,缺血2 h后拔出线栓造成缺血再灌注损伤(ischemia reperfusion injury,I/R)。TTC染色法观察丹灯通脑胶囊对脑梗死体积的影响。激光多普勒血流仪(laser doppler flowmetery,LDF)测定缺血再灌注大鼠局部脑血流量;采用苏木素伊红(hematoxylin and eosin, HE)染色、透射电镜(transmission electron microscopy, TEM)观察大鼠海马神经元损伤;微透析系统检测脑缺血后氨基酸含量变化;流式分析预防治疗脑内海马钙离子含量。结果 结果显示,与模型组相比,丹灯通脑胶囊组大鼠7、14 d脑血流量回升且脑梗死体积下降;海马CA1区神经元损伤明显减轻;脑内兴奋性氨基酸含量和海马内钙离子含量降低。结论 提示丹灯通脑胶囊有明显预防和治疗大鼠脑缺血再灌注损伤,该保护作用可能与抑制兴奋性氨基酸释放及减少胞内钙离子有关。
Abstract
OBJECTIVE To observe the protective effects of Dandeng Tongnao capsule (DDTN) on cerebral ischemia reperfusion injury in rat and explore its mechanism so as to reevaluation the clinical efficacy of DDTN. METHODS Middle cerebral artery occlusion (MCAO) model in rats was established using filament method and reperfusion was processed with filament removed 2 h later. Therefore, the MCAO models were established to observe the effect of DDTN-L group (0.21 g·kg-1) and DDTN-H group (0.84 g·kg-1) on infract volume via TTC staining. Regional cerebral blood flow(rCBF) of corpus striatum of the side of cerebral infraction was measured by using laser doppler flow (LDF). Pathological changes of neuron in hippocampuses were observed via hematoxylin and eosin (HE) staining and transmission electron microscopy (TEM).The changes of amino acid in cerebral ischemia were detected by microdialysis system. The level of Ca2+ was evaluated by flow cytometer (FCM). RESULTS The results showed that the cerebral blood flow increased and infract volume reduced at 7, 14 d in DDTN group. Analysis of HE staining and TEM scan suggested that DDTN (0.21, 0.84 g·kg-1) remarkably alleviated neuronal injury of hippocampal CA1 area during I/R. The contents of Ca2+ and excitatory amino acids were significantly decreased in DDTN group. CONCLUSION DDTN could prevent cerebral injury of MCAO rats via decreasing the intracellular Ca2+ concentration, and inhibiting the release of excitatory amino acids.
关键词
丹灯通脑胶囊 /
脑缺血 /
微透析 /
局部血流量 /
激光多普勒血流仪
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Key words
Dandeng-Tongnao granule /
cerebral ischemia /
microdialysis /
regional cerebral blood flow /
laser doppler flow
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中图分类号：
R965
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参考文献
[1] National Health and Family Planning Commission Stroke Screening and Prevention Engineering Committee. Stroke screening and prevention norms[J]. Chin J Front Med Sci(中国医学前沿杂志电子版),2013,5(9):44.
[2] Guidelines for the diagnosis and treatment of acute ischemic stroke in the cerebrovascular group of the Chinese Medical Association Neurology Branch. Guidelines for the diagnosis and treatment of acute ischemic stroke in China[J]. Chin J Front Med Sci(中国医学前沿杂志电子版),2010,2(4):16.
[3] LONGA E Z,WEINSTEIN P R,CARLSON S, et al. Reversible middlecerebral artery occlusion without craniectomy in rats[J]. Stroke,1989,20(1):84.
[4] BENVENISTE H, DREJER J, SCHOUSBOE A, et al. Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracerebralmicrodiaiysis[J]. Neurochem,1984,43(5):1369-1374.
[5] HARADA H, WANG Y, MISHIMA Y, et al. A novel method of detecting rCBF with laser-Doppler flowmetry without cranial window through the skull for a MCAO rat model[J]. Brain Res Protoc,2005,14(3):165-170.
[6] SCHMID-ELSAESSER R, ZAUSINGER S, HUNGERHUBER E, et al. A critical reevaluation of the intraluminal thread model of focal cerebral ischemia:evidence of inadvertent premature reperfusion and subarachnoid hemorrhage in rats by laser-Doppler flowmetry[J]. Stroke,1998,29(10):2162-2170.
[7] LI F, ZHENG G C, QU Y P, et al. Clinical application of Dan Deng Tong Nao Capsule[J]. Chin J Ethnomed Ethnopharm (中国民族民间医药),2016,25(13):35-37.
[8] ZHANG J H, HUANG C, ZHANG Z N, et al. Clinical observation on 30 cases of cerebral infarction recovering with Dandeng Tongnao capsule[J]. Guide J Tradit Chin Med Pharm (中医药导报),2014,20 (7):109-110.
[9] WEI W H, BAI L, SI X, et al. Clinical observation of Dandeng Tongnao capsule in treating the causes of cerebral infarction[J]. Chin J Pract Nervous Dis (中国实用神经疾病杂志),2012,15 (5):51-52.
[10] PANG X, LI T, FENG L, et al. Ellagic acid-induced thrombotic focal cerebral ischemic model in rats[J]. J Pharmacol Toxicol Methods,2014,69: 217-222.
[11] ARRANZ A M, GOTTLIEB M, PEREZ-CERDA F, et al. Increased expression of glutamate transportersin subcortical white matter after transient focal cerebral ischemia[J]. Neurobiol Dis,2010,37(1): 156-165.
[12] ZHANG Y, CHEN Q, GU W D, et al. Effects of erigeron on ATP content and release of excitatory amino-acids of the hippocampus after cerebral ischemia/reperfusion injury in gerbils[J]. Jiangsu Med J(江苏医药),2005,31(11):843-845.
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脚注
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基金
北京市科技计划课题资助(Z161100002616024)
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