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  

Angiotensin 1-7 Protects against Oxygen-Glucose Deprivation /Reoxygenation-induced Injury of Primarily Cultured Hippocampal Neurons of Rats

作　　者：卞伟华[1] 陈明迎 张小华[3] 许聪[1] 孙业盈[4] 杨玥[1] 杨美子[5] 

机构地区：[1]滨州医学院细胞生物学教研室,山东烟台264003 [2]广东省连州卫生学校教务科,广东连州513400 [3]滨州医学院生物技术教研室,山东烟台264003 [4]滨州医学院遗传教研室,山东烟台264003 [5]滨州医学院药理学教研室,山东烟台264003

出　　处：《中国生物化学与分子生物学报》2016年第10期1149-1154,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：山东省自然科学基金(No.ZR2013CL004;No.ZR2012HM057;No.ZR2013CL003);泰山学者建设工程专项经费资助~~

摘　　要：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 important roles in the nervous system. Previous studies found that Ang1-7 played a protective role in cerebral ischemia animal model. But the protection effect of Ang1-7 on hippocampal neuron injury induced by oxygen-glucose deprivation/reoxygenation and its mechanism had not been reported. In this study,the hippocampal neuron of new born SD rat was cultivated. Anaerobic incubator and EBSS medium were used to establish oxygen-glucose deprivation /reoxygenation model of hippocampal neuron. The cells were divided into three groups： the normal control group,the experimental contro1 group and the Ang1-7 stimulation group. Inverted microscope was adopted to observe the morphology of hippocampal neurons. Compared with the experimental control group,the cell morphology in Ang1-7 group was greatly improved. The results of CCK8 assays showed that the neuron vitality in Ang1-7 group was greatly increased. Flow cytometry was adopted to study cell apoptosis and the levels of Ca2 ＋and NO in hippocampal neurons. The results showed that cell apoptosis and necrosis rate in Ang1-7 group were decreased. At the same time,the levels of Ca2 ＋and NO in Ang1-7 group were significantly decreased. Western blot showed that Bax expression was decreased and Bcl-2expression was increased in Ang1-7 group. These results suggested that Ang1-7 played protective roles through increasing Bcl-2 expression,reducing the levels of Ca2 ＋,NO and Bax,and reducing cell apoptosis and necrosis rate. The results in this study would lay the foundation for further research on the protection mechanism of Ang1-7 in neurons and the prevention and control of cerebral ischemia related diseases.

关 键 词：血管紧张素1-7 海马神经元 氧糖剥夺/复氧 凋亡 

分 类 号：Q279[生物学—细胞生物学]