摘要
目的研究丹皮酚对大鼠神经元缺糖缺氧再灌损伤的保护作用,并探讨其作用机制。方法建立原代培养的新生大鼠神经元缺糖缺氧再灌损伤模型,随机分组为:对照组;模型组;丹皮酚组:建立模型,每个再灌时间点均经0.2×10-6,1×10-6,5×10-6mol·L-1 3个浓度药物处理;MK-801阳性对照药组。倒置相差显微镜下进行一般形态学观察,用MTT法测定神经元存活率。通过放射配基结合实验测定N-甲基-D-天门冬氨酸受体(NMDA受体)结合力,采用荧光法测定细胞内游离钙离子([Ca2+]i)浓度及应用试剂盒测定一氧化氮(NO)含量及一氧化氮合酶(NOS)活性。结果丹皮酚可显著降低大鼠皮质和海马神经元缺糖缺氧再灌损伤时细胞死亡率,减弱NMDA受体结合力,降低神经细胞内游离钙离子浓度([Ca2+]i)及NO含量、NOS活性的升高。结论丹皮酚对离体培养的大鼠神经元缺糖缺氧再灌损伤具有明显保护作用,其作用机制可能与抗氧化及减弱NMDA受体结合力有关。
Abstract
OBJECTIVE To determine if paeonol can protect cultured rat neurons from anoxia-induced injury and elucidate the underlying mechanism.METHODS The rat neurons cultivated as the object of study and then build the model of oxygen-glucose deprivation and reperfusion.They were randomly divided into 4 groups: control group;injury model group;paeonol treatment groups,the paeonol was added before reperfusion,which included the three concentration of paeonol(0.2×10-6mol·L-1,Pae-L;1×10-6mol·L-1,Pae-M and 5×10-6mol·L-1,Pae-H);MK-801 group(10×10-6mol·L-1).Each group was observed by inverted phase contrast microscope;neuron viability was measured by the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT);binding force of NMDA receptors was evaluated by liquid scintillation counting;intracellular Ca2+ level was determined by fluorospectrophotometer.NO content and NOS activity were observed.RESULTS Compared with OGD/reperfusion group at each designated period,paeonol treatment after OGD obviously increased cell survival rate and reduced the binding force of NMDA receptors,decreased the activity of NOS and the content of NO and intracellular Ca2+.CONCLUSION Paeonol can protect ret neurons from ischemia-reperfusion injury by alleviating morphological damage and increasing neuron viability.
关键词
丹皮酚 /
缺糖缺氧损伤 /
NMDA受体 /
氧自由基 /
缺血性脑血管病
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Key words
paeonol /
oxygen-glucose deprivation /
NMDA receptor /
oxygen free radical /
ischemic cerebral vascular disease
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宋宁宁;魏欣冰;刘睿;武继彪;张岫美.
丹皮酚对原代培养的大鼠皮质和海马神经元缺糖缺氧再灌损伤的保护作用[J]. 中国药学杂志, 2007, 42(05): 353-356
SONG Ning-ning;WEI Xin-ing;LIU Rui;WU Ji-io;ZHNG Xiu-mei.
Prorective Effects of Paeonol on Cultured Cortical and Hippocampal Neurons Subjected to Oxygen-Glucose Deprivation and Reperfusion in Rats [J]. Chinese Pharmaceutical Journal, 2007, 42(05): 353-356
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脚注
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基金
国家自然科学基金资助项目(30572187)
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