血管紧张素1-7对氧糖剥夺/复氧的原代培养大白鼠海马神经元的保护作用

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血管紧张素1-7(angiotensin 1-7,Ang1-7)在神经系统中发挥重要作用。已有研究发现,Ang1-7在脑缺血动物模型中发挥保护作用,但至今未见有关Ang1-7对氧糖剥夺/复氧(oxygenglucose deprivation/reoxygenation,OGD/R)损伤神经元的保护作用及其机制的研究报道。本研究以厌氧培养及不含葡萄糖的EBSS培养基培养、建立新生大白鼠原代培养的海马神经元OGD/R模型模拟脑缺血环境,实验分为3组:正常对照组、实验对照组和Ang1-7处理组。倒置显微镜观察神经元形态显示,Ang1-7处理组的神经元形态明显改善;CCK8试剂盒检测发现,Ang1-7处理组的细胞活性提高;流式细胞术研究发现,Ang1-7处理组的神经元凋亡和坏死率降低、神经元内Ca2+及NO水平降低;Western印迹结果发现,Ang1-7处理组Bax表达降低,Bcl-2表达增加。以上结果说明,Ang1-7可降低OGD/R神经元中NO和Ca2+水平,降低Bax蛋白、增加Bcl-2蛋白的表达,减少OGD/R神经元凋亡和坏死率,对OGD/R神经元发挥了保护作用。本研究为进一步在神经元水平上研究Ang-1-7的保护机制奠定基础,对中风等脑缺血疾病的防治具有重要意义。 Angiotensin 1-7 (Ang1-7) plays an important role in the nervous system. It has been found that Ang1-7 plays a protective role in animal models of cerebral ischemia, but so far no protective effect of Ang1-7 on neurons damaged by oxygenglucose deprivation / reoxygenation (OGD / R) has been found And its mechanism of research reports. In this study, anaerobic and glucose-free EBSS culture medium was cultured to establish a newborn rat primary cultured hippocampal neurons OGD / R model to simulate cerebral ischemia environment, the experiment is divided into 3 groups: normal control group, experimental control group And Ang1-7 treatment group. The morphology of neurons was observed by inverted microscope. The morphology of neurons in Ang1-7 group was significantly improved. The CCK8 kit assay showed that Ang1-7-treated cells showed an increase in cell viability. Flow cytometry showed that Ang1-7- Apoptosis and necrosis rates were decreased and Ca2 + and NO levels in neurons were decreased. Western blotting showed that Ang1-7 treatment decreased Bax expression and Bcl-2 expression. The above results indicate that Ang1-7 can reduce the levels of NO and Ca2 + in OGD / R neurons, decrease Bax protein, increase the expression of Bcl-2 protein and decrease the apoptosis and necrosis rate of OGD / R neurons. OGD / R neurons Play a protective role. This study lays the foundation for further study on the protective mechanism of Ang-1-7 at the neuron level and is of great significance for the prevention and treatment of stroke and other cerebral ischemic diseases.
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