EPO对缺氧缺糖受损神经细胞的作用研究

发布时间：2018-11-20 20:04

【摘要】：目的：建立缺氧缺糖神经细胞损伤模型，探讨EPO发挥神经保护作用的可能机制，希望为EPO在神经系统疾病的临床应用提供理论依据。方法：将出生后24h内C57BL/6小鼠的大脑皮质神经元原代培养7-10d后，神经元特异性烯醇化酶（Neuron specific enolase, NSE）免疫组化染色方法鉴定神经元的纯度；用含连二亚硫酸钠的无糖Earle’s液模拟大脑缺血缺氧性损伤，建立缺氧缺糖神经细胞损伤模型。分组：1、正常组；2、模型组（缺氧缺糖神经细胞损伤模型）；3、EPO组(EPO组干预的缺氧缺糖神经细胞损伤模型）。MTT检测细胞存活率、FITC-Annexin V/PI荧光染色方法检测细胞凋亡率、测定LDH漏出率，观察EPO对缺氧缺糖损伤神经元的保护作用。结果：1、成功原代培养神经细胞，经NSE免疫组化方法鉴定神经元纯度大于90%；2、建立缺氧缺糖神经细胞损伤模型，模型组中神经细胞凋亡率为（15.2±0.37），LDH漏出率为（20.4±0.33），细胞存活率为（54.1±0.51），与正常组相比差异有统计学意义（P0.01）；3、与模型组相比，EPO组的细胞凋亡率、LDH漏出率低于模型组，细胞存活率高于模型组（P0.01）。结论：原代培养小鼠神经细胞，成功构建缺氧缺糖神经细胞损伤模型。EPO能够减轻缺氧缺糖对神经元的损伤，，该保护作用可能是通过抑制神经细胞凋亡途径介导完成。
[Abstract]:Objective: to establish a model of hypoxia-glucose deficiency nerve cell injury and to explore the possible mechanism of neuroprotective effect of EPO in order to provide a theoretical basis for the clinical application of EPO in nervous system diseases. Methods: after primary culture of cerebral cortical neurons of C57BL/6 mice within 24 hours after birth for 7-10 days, the purity of neurons was identified by immunohistochemical staining with neuron-specific enolase (Neuron specific enolase, NSE). A model of hypoglycemic neuronal injury was established by simulating cerebral ischemia and hypoxia injury with Earle's solution containing sodium bisulfite. Groups: 1, normal group, 2, model group (hypoxia and glucose deficiency nerve cell injury model); (3). MTT, FITC-Annexin V/PI fluorescence staining and LDH leakage were used to detect cell survival rate, apoptosis rate and LDH leakage rate in EPO group (hypoxia / glucose deficient nerve cell injury model). To observe the protective effect of EPO on neurons injured by hypoxia and glucose deficiency. Results: 1. The primary cultured neurons were successfully cultured, and the purity of neurons was higher than 90 by NSE immunohistochemical method. 2. In the model group, the apoptosis rate was (15.2 卤0.37), LDH leakage rate was (20.4 卤0.33), and the cell survival rate was (54.1 卤0.51). Compared with the normal group, the difference was statistically significant (P0.01). 3Compared with the model group, the apoptosis rate and LDH leakage rate in EPO group were lower than those in the model group, and the cell survival rate was higher than that in the model group (P0.01). Conclusion: EPO can attenuate the neuronal damage induced by hypoxia and glucose deficiency, which may be mediated by inhibiting neuronal apoptosis.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R741

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