不同时长氧糖剥夺对大鼠原代皮层神经元-小胶质细胞共培养体系的影响  

Effects of Oxygen and Glucose Deprivation for Different Time on Primary Cortical Neuron-Microglia Co-culture System of Rats

作　　者：程伟能 张玉琴[1] 赖文芳[1] 陈亚萍 南丽红[1] 黄枚[1] Cheng Weineng;Zhang Yuqin;Lai Wenfang;Chen Yaping;Nan Lihong;Huang Mei(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122)

机构地区：[1]福建中医药大学药学院,福州350122

出　　处：《中药药理与临床》2021年第4期195-200,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:81873031);福建中医药大学校管科研课题(编号:X2020008-重点)。

摘　　要：目的:探讨不同时长氧糖剥夺(OGD)对大鼠原代皮层神经元-小胶质细胞共培养体系的影响。方法:提取新生SD大鼠原代皮层神经元及小胶质细胞,经免疫荧光法鉴定细胞纯度达到95%以上,可用于建立大鼠原代皮层神经元-小胶质细胞共培养体系并进行OGD处理;分别设置空白对照组、氧糖剥夺0.5 h、1 h、2 h、4 h组,除空白对照组不进行OGD处理,其余各组分别OGD处理0.5、1、2、4 h;复糖复氧24 h后,比色法检测上清中乳酸脱氢酶(LDH)含量,TUNEL法检测细胞凋亡率,免疫荧光法检测离子钙结合衔接分子1(Iba1)阳性细胞数,ELISA法检测上清中肿瘤坏死因子-α(TNF-α)含量。结果:与空白对照组相比,氧糖剥夺4 h组细胞凋亡率、Iba1阳性细胞率无显著性差异;氧糖剥夺0.5 h组、1 h组和2 h组中的LDH含量、细胞凋亡率、Iba1阳性细胞数及TNF-α含量均明显增多(P<0.05或P<0.01)。与氧糖剥夺2 h组相比,氧糖剥夺0.5 h组中的LDH含量及TNF-α含量明显降低;氧糖剥夺1 h组中TNF-α含量明显降低(P<0.05或P<0.01)。结论:大鼠原代皮层神经元-小胶质细胞共培养体系经OGD处理2 h可较好模拟缺血性脑损伤后两细胞的相互作用。Objective:To investigate the effects of oxygen and glucose deprivation(OGD) for different time on primary cortical neuron-microglia co-culture system of rats. Methods: Primary cortical neurons and microglial cells of newborn SD rats were extracted and identified for over 95% purity by immunofluorescence. The primary cortical neuron-microglia co-culture system was established and cultured under OGD. Cells were divided into a blank control group, an OGD 0.5 h group, an OGD 1 h group, an OGD 2 h group, and an OGD 4 h group. The cells in the OGD groups were subjected to OGD treatment for 0.5 h, 1 h, 2 h, and 4 h, respectively. After 24 hours of reoxygenation, the content of lactate dehydrogenase(LDH) in the supernatant was detected by colorimetry. The apoptosis rate was detected by TUNEL assay. The number of ionized calcium-binding adapter molecule 1(Iba1) positive cells was detected by immunofluorescence. The content of tumor necrosis factor-α(TNF-α) in the supernatant was detected by ELISA. Results:Compared with the blank control group, the OGD 4 h group showed insignificant difference in the apoptosis rate and the number of Iba1 positive cells, while other OGD groups displayed increased LDH content, apoptosis rate, number of Iba1 positive cells, and TNF-α content(P<0.05 or P<0.01). Compared with the OGD 2 h group, the OGD 0.5 h group showed declining LDH and TNF-α levels, while the OGD 1 h group showed reduced TNF-α level(P<0.05 or P<0.01). Conclusion: The rat primary cortical neuron-microglia co-culture system treated with OGD for 2 h can simulate the interaction of the two cells after ischemic brain injury.

关 键 词：氧糖剥夺 神经元 小胶质细胞 凋亡 乳酸脱氢酶 离子钙结合衔接分子1 肿瘤坏死因子-Α 

分 类 号：R743.3[医药卫生—神经病学与精神病学]