摘要
目的:观察丹红注射液对脑缺血再灌注损伤大鼠和缺氧损伤神经元细胞的保护作用。方法:①成年雄性Wistar大鼠随机分为模型组、假手术组、丹红注射液(0.9,1.8,3.6 mL·kg-1)组和阳性对照(依达拉奉,6 mg·kg-1)组,采用线栓法制备大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)模型,缺血60 min后再灌注72 h,分别于造模前30 min、再灌注即刻、缺血后24,48,72 h经ip给药。72 h,进行神经功能评分;末次给药1 h后,取左侧缺血半脑,匀浆,ELISA法测定S100B蛋白和神经特异烯醇化酶(NSE)含量。②体外培养Neuro-2A细胞,分为正常对照组、氧糖剥夺(oxygen and glucose deprivation,OGD)损伤模型组、丹红注射液组,细胞加平衡盐溶液D-Hanks于94%N2-5%CO2-1%O2三气培养箱缺氧孵育2 h建立OGD损伤模型,丹红注射液(1.25,2.5,5 mL.L-1)干预处理,检测细胞的增殖活力、乳酸脱氢酶(LDH)漏出量、活性氧(ROS)水平。结果:①与假手术组比,模型组大鼠神经功能评分明显升高(P<0.01),脑组织匀浆NSE和S100B的含量明显升高(P<0.01),与模型组比,丹红注射液各剂量组均能不同程度改善MCAO大鼠的行为障碍(P<0.05,P<0.01,P<0.05),明显降低脑组织匀浆中NSE含量(P<0.05),中、低剂量组明显降低S100含量(P<0.05,P<0.01)。②与正常对照组比,模型组细胞活力明显下降,LDH漏出量升高,ROS水平显著升高(P<0.01),与OGD组比,丹红注射液各组均能增加OGD损伤Neuro-2A细胞活力,降低细胞内ROS水平(P<0.01),2.5,5 mL.L-1浓度丹红注射液显著降低细胞LDH漏出量(P<0.05,P<0.01)。结论:丹红注射液对脑缺血缺氧损伤具有一定的保护作用。
Objective：To investigate the protective effect of Danhong injection on cerebral ischemiareperfusion injury in rats and hypoxic injury in Neuro-2A cell.Method：①Adult male Wistar rats were randomly divided into 6 groups as following：model group,sham group,Danhong injection low,middle and high dose（0.9,1.8,3.6 mL·kg-1） groups,and the positive control（edaravone,6 mg·kg-1） group.The model of focal cerebral ischemia-reperfusion injury was reproduced by middle cerebral artery occlusion（MCAO） performed with thread inserted for 60 minutes and then reperfusion 72 h,Danhong injection was administered intraperitoneally to the animals 30 min before operation and immediately at the onset of reperfusion,and then once a day for three days.Scoring method of ischemia for 72 h was used to evaluate the abnormal behaviour by nerve injury.The content of S100B protein and nerve specific enolase（NSE） in brain tissue were determined by enzyme linked immunoassay（ELISA）.②Neuro-2A cells injury were reproduced by oxygen and glucose deprivation（OGD）.Incubating with Danhong injection（1.25,2.5,5 μL.mL-1）,the neuroprotective effect indexes were investigated as the cell proliferation activity,LDH release,and ROS detection.Result：① Compared with sham group,the contents of S100B and NSE were significantly increased in brain tissue（P ＆lt; 0.01）.Compared with model operation group,Danhong injection 0.9,1.8,3.6 mL·kg-1 ip could improve the nerve defect symptoms of model rats at 72 h to some extent（P ＆lt; 0.05,P ＆lt; 0.01,P ＆lt; 0.05）,and decreased the content of NSE significantly（P ＆lt; 0.05）.Danhong injection 0.9,1.8 mL·kg-1 ip could markedly decrease the content of S100B protein（P ＆lt; 0.05,P ＆lt; 0.01）.② Compared with control group,the viability of Neuro-2A cell injury induced by OGD decreased significantly,the release of LDH and the level of ROS significantly increase（P ＆lt; 0.01）,1.25,2.5,5 mL.L-1Danhong injection could improve hypoxic injury Neuro-2A cell proliferative activity,inhibit the level of ROS increase in cells（P ＆lt; 0.01） and 2.5,5 mL.L-1Danhong injection could reduce the LDH release of Neuro-2A cell（P ＆lt; 0.05,P ＆lt; 0.01）.Conclusion：Danhong injection protect against cerebral the hypoxic-ischemic injury.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第17期221-224,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家"重大新药创制"科技重大专项(2012ZX09101201-004)
天津市科技创新体系及条件平台建设(10SYSYJC28900)
中药有效成分群辨识技术研究(2008BAI51B01)
作者简介
李美娇，硕士，从事中药药理研究，Tel：18920011537，E—mail：meijiaojia0851020@163．com
【通讯作者】胡利民，博士，研究员，从事脑血管病中药药理研究，Tel：022~9596166，E-mail：huliminth@126．com
