香豆素衍生物瑞香素对HT22细胞中谷氨酸毒性损伤及脑缺血损伤的保护作用研究

发布时间：2018-04-23 09:12

本文选题：瑞香素 + 脑缺血再灌注　；参考：《苏州大学》2014年硕士论文

【摘要】：第一部分瑞香素（Daphnetin）对小鼠大脑中动脉缺血再灌注损伤及新生大鼠缺氧缺血性损伤的保护作用目的：观察瑞香素(Daphnetin, DAP)对小鼠大脑中动脉缺血再灌注损伤及新生大鼠缺氧缺血性损伤是否具有保护作用。方法：将雄性ICR小鼠及新生SD大鼠随机分为安慰剂(sham)组、安慰剂+DAP组、安慰剂+缺血再灌注(ischemia/reperfusion, I/R)(or缺氧缺血(hypoxia/ischemia, H/I))组和DAP+I/R(or H/I)组，建立小鼠短暂大脑中动脉阻塞(transient middle cerebral arteryocclusion, tMCAO)模型及新生大鼠缺氧缺血(H/I)模型，观察DAP的处理对小鼠脑梗死体积、神经行为功能及对新生大鼠缺氧缺血性脑梗死的影响。结果：与对照组相比，经DAP处理后小鼠的脑梗死体积明显减小，神经行为功能明显增强(P0.05)。缺血再灌注前侧脑室预给予DAP(1mg/kg)，脑梗死体积减小了24.6%；神经行为学评分具有明显统计学差异(P0.05)。同样，缺血再灌注后不同时间段分别予以腹腔注入DAP(10mg/kg)，与对照组相比，经DAP预处理后的新生大鼠的脑梗死体积明显缩小，甚至缺血4小时后给药仍有保护作用(P0.05)。结论：使用DAP干预小鼠tMCAO模型及新生大鼠H/I模型，小鼠的脑梗死体积明显减小，，神经行为功能缺损明显减轻；新生大鼠的脑梗死体积明显减少。这些结果均提示DAP对于小鼠tMCAO及新生大鼠H/I诱导的脑缺血损伤具有神经保护作用。第二部分瑞香素（Daphnetin）对HT22细胞中谷氨酸毒性损伤的保护作用的相关机制目的：研究瑞香素(Daphnetin, DAP)对HT22细胞中谷氨酸毒性的保护及对谷胱甘肽(Glutathione, GSH)及超氧化物歧化酶(superoxide dismutase, SOD)的影响。探讨DAP神经保护作用的相关机制。方法：将HT22细胞培养过夜，后加入5mmol/L谷氨酸制作神经元损伤模型组。培养基中加入不同浓度的DAP与培养基的混合液共同孵育12小时，后在显微镜下分别观察各组神经元形态变化；采用MTS法检测细胞活力；测定细胞内GSH含量及SOD活力的变化。结果： DAP能有效地提高HT22细胞的存活率，其最佳保护浓度是100μmol/L。以5mmol/L谷氨酸作用于培养HT22细胞，细胞形态表现为突起减少。应用DAP100μmol/L后可改善因谷氨酸引起的HT22细胞形态的改变，可维持谷氨酸损伤后HT22细胞内GSH的含量及SOD的活性。结论： DAP可明显拮抗谷氨酸毒性作用。其可能的机制与DAP维持GSH含量及SOD活性有关。
[Abstract]:The Protective effect of Daphnetin on Middle Cerebral artery Ischemia-reperfusion injury in mice and Hypoxic-Ischemic injury in Neonatal Rats Aim: to observe the protective effect of daphnetin (DAP1) on middle cerebral artery ischemia reperfusion injury in mice and hypoxic ischemic injury in newborn rats. Methods: male ICR mice and newborn SD rats were randomly divided into three groups: the placebo group, the placebo DAP group, the placebo DAP group, the placebo ischemia reperfusion group, the I/R)(or hypoxic ischemia hypoxia-r-ischemic ischemic reperfusion group, and the DAP I/R(or ischemia reperfusion group. The transient middle cerebral arterial occlusion (tMCAO) model of transient middle cerebral artery occlusion (TMCAO) in mice and the model of hypoxic-ischemic H / I in neonatal rats were established to observe the effects of DAP treatment on cerebral infarction volume, neurobehavioral function and hypoxic-ischemic cerebral infarction in neonatal rats. Results: compared with the control group, the volume of cerebral infarction and the neurobehavioral function of the mice treated with DAP were significantly decreased and the neurobehavioral function was significantly enhanced (P 0.05). The volume of cerebral infarction decreased by 24.6mg 路kg ~ (-1) 路kg ~ (-1) and the neurobehavioral score was significantly different (P _ (0.05) ~ 0. 05) when the anterior ventricle was given 1 mg 路kg ~ (-1) of DAPG / kg after ischemia-reperfusion. Similarly, 10 mg / kg DAP was injected intraperitoneally at different time points after ischemia / reperfusion. Compared with the control group, the volume of cerebral infarction in neonatal rats pretreated with DAP was significantly reduced, and even after 4 hours of ischemia, the drug still had a protective effect (P 0.05). Conclusion: the cerebral infarct volume and neurobehavioral deficit were significantly decreased and the cerebral infarction volume of newborn rats was significantly decreased after DAP was used to interfere with tMCAO model and neonatal rat model of H / P I. These results suggest that DAP has neuroprotective effects on cerebral ischemia injury induced by tMCAO in mice and H / P I in newborn rats. Part two the protective mechanism of daphnetin on glutamate toxicity in HT22 cells Aim: to study the protection of daphnetin (DAPs) against glutamate toxicity in HT22 cells and its effects on glutathione (GSH) and superoxide dismutase (sod). To explore the mechanism of neuroprotective effect of DAP. Methods: HT22 cells were cultured overnight and 5mmol/L glutamate was added to model group. Different concentrations of DAP were added to the culture medium for 12 hours, then the morphological changes of neurons were observed under microscope, the cell viability was detected by MTS assay, and the changes of GSH content and SOD activity in the cells were measured. Results: DAP could effectively improve the survival rate of HT22 cells, and the best protective concentration was 100 渭 mol / L. When 5mmol/L glutamate was used to culture HT22 cells, the morphology of the cells decreased. DAP100 渭 mol/L could improve the morphological changes of HT22 cells induced by glutamate, and maintain the content of GSH and the activity of SOD in HT22 cells after Glutamic acid injury. Conclusion: DAP can antagonize the toxicity of glutamate. The possible mechanism is related to the maintenance of GSH content and SOD activity by DAP.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R743.31

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