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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 observe the protective effect of Danhong injection on neuronal cells with hypoxic injury after cerebral ischemia-reperfusion injury in rats. Methods: ① Adult male Wistar rats were randomly divided into three groups: model group, sham operation group, Danhong injection group (0.9,1.8,3.6 mL · kg -1) and positive control group (edaravone 6 mg · kg -1 ) Group. Middle cerebral artery occlusion (MCAO) model was established by thread occlusion method. The rats were reperfused for 60 min and then reperfused for 72 h at 30 min before reperfusion and immediately after reperfusion , 48,72 h via ip administration. The neurological function score was measured at 72 h after the last administration. The levels of S100B protein and neuron specific enolase (NSE) were determined by ELISA. ② Neuro-2A cells were cultured in vitro and divided into normal control group, OGD injury model group, Danhong injection group and D-Hanks cells with balanced salt solution in 94% N2-5% OGD injury model was established by incubating CO2-1% O2 three times in hypoxia for 2 hours. The cells were treated with Danhong injection (1.25, 2.5 and 5 mL.L-1) for 24 hours. The activity of lactate dehydrogenase ) Leakage, reactive oxygen species (ROS) levels. Results: ①Compared with the sham operation group, the neurological scores of the model group were significantly increased (P <0.01), and the levels of NSE and S100B in the brain tissue homogenate were significantly increased (P <0.01). Compared with the model group, (P <0.05, P <0.05), and significantly decreased the content of NSE in brain tissue homogenate (P <0.05), middle and low dose group Significantly reduce S100 content (P <0.05, P <0.01). Compared with the normal control group, the viability of the model group decreased significantly, the leakage of LDH increased and the level of ROS increased significantly (P <0.01). Compared with the OGD group, the Danhong injection group can increase Neuro-2A (P <0.01). 2.5 and 2.5 mL.L-1 Danhong injection significantly reduced the amount of LDH leakage (P <0.05, P <0.01). Conclusion: Danhong injection has a protective effect on cerebral ischemia and hypoxia injury.