原代大鼠星形胶质细胞对氧糖剥夺的增殖性应答反应  

Proliferative Response of Astrocytes to Oxygen-Glucose Deprivation in Primary Rats

作　　者：范玉颖[1] 张俊梅[1] 王华[1] 赵晓春[2] 

机构地区：[1]中国医科大学附属盛京医院小儿神经内科,沈阳110004 [2]中国医科大学附属盛京医院麻醉科,沈阳110004

出　　处：《实用儿科临床杂志》2012年第20期1596-1599,共4页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金青年科学基金(81000824)

摘　　要：目的建立良好的模拟缺氧缺血性脑损伤后星形胶质细胞增殖的体外模型的方法。方法采用CCK-8(Cell CountingKit-8)法测定星形胶质细胞活力,5-乙炔基-2'脱氧尿嘧啶核苷(EdU)掺入法检测星形胶质细胞DNA合成,免疫组织化学染色法检测星形胶质细胞增殖细胞核抗原(PCNA)表达。比较原代大鼠星形胶质细胞在氧糖剥夺(OGD),10 mL·L-1氧气、无糖含血清培养基)培养不同时间(0 h、3 h、6 h、9 h)的增殖情况。结果 CCK-8检测结果显示随OGD培养时间延长,原代大鼠星形胶质细胞活力逐渐增强,6 h达高峰,与相应时间点正常培养组比较差异有统计学意义;随后减弱,9 h已接近正常培养组水平。OGD培养后,原代大鼠星形胶质细胞EdU和PCNA阳性细胞率逐渐增加,6 h达最高水平,与相应时间点的正常培养组比较差异有统计学意义;而后下降,至9 h与正常培养组比较差异无统计学意义。结论 OGD培养6 h可成功模拟体内缺氧缺血后星形胶质细胞增殖改变,为进行缺氧缺血性脑损伤后星形胶质细胞过度增殖的相关研究提供了稳定可靠的体外实验模型和理论依据。Objective To explore the method for building a good in vitro model mimicking astrogliosis after hypoxic - ischemic brain in-jury during the neonatal period. Methods The vitalities of astrocytes of primary rats were measured by adopting cell counting kit - 8 （ CCK - 8 ） method. Ethynyl deoxyuridine （EdU） incorporation was used to test DNA synthesis of the astrocytes, and immunohistochemistry staining was used to test the proliferating cell nuclear antigen （PCNA） of astrocytes. The proliferations of the astrocytes of primary rats at different time points （0 h,3 h ,6 h and 9 h） after oxygen-glucose deprivation （OGD）, 10 mL · L^-1 oxygen, dulbecco＇s minimum essential medium without glucose） were compared. Results With OGD cultivation prolonged, the vigor of astrocytes of primary rats increased and reached to the highest level at OGD 6 h by CCK-8 detection, compared with normal control group, at the corresponding time point, and the differences were significant. Afterwards the vigor decreased, at OGD 9 h, and the vigor of astrocytes of primary rats decreased nearly to the level of the control group. After OGD cultivation, the EdU and PCNA positive ratio of astrocytes of primary rats increased gradually to the most extent at OGD 6 h; compared with normal control group at the corresponding time point, the differences were significant. Afterwards the vigor decreased, at 9 h, the differences, compared with the normal cultivating control group, had no statistical significance. Conclusions The OGD cultivation for 6 h can successfully mimic the astrogliosis after hypoxic - ischemia brain injury, and this can provide a stable in vitro experimental model and theoreti-cal basis for correlational studies on excessive astrocyte proliferation after hypoxic - ischemia brain injury.

关 键 词：星形胶质细胞 氧糖剥夺 缺氧缺血 增殖 

分 类 号：R748[医药卫生—神经病学与精神病学]