DHA对氧糖剥夺-复氧复糖损伤时小胶质细胞活化的影响  

Effect of docosahexaenoic acid on microglial activation during oxygen-glucose deprivation and restoration injury

作　　者：冯燕[1] 王魁 张霞婧[1] 邵勇平[1] 赵蔚[1] Feng Yan;Wang Kui;Zhang Xiajing;Shao Yongping;Zhao Wei(Department of Anesthesiology,Xi′an No.4 Hospital,Xi′an 710001,China;Department of Anesthesiology,Second Affiliated Hospital,Xi′an Jiaotong University,Xi′an 710004,China)

机构地区：[1]西安市第四医院麻醉科,710001 [2]西安交通大学第二附属医院麻醉科,710004

出　　处：《中华麻醉学杂志》2019年第8期928-930,共3页Chinese Journal of Anesthesiology

摘　　要：目的评价二十二碳六烯酸(DHA)对氧糖剥夺-复氧复糖损伤时小胶质细胞活化的影响。方法将N9小胶质细胞以104个/孔的密度接种于96孔细胞培养板培养3~5 d,采用随机数字表法分为3组(n=18):对照组(C组)、氧糖剥夺-复氧复糖组(OGD/R组)和DHA+氧糖剥夺-复氧复糖组(DHA+OGD/R组)。C组37℃培养箱(95%空气-5%CO2)中培养24 h;OGD/R组和DHA组将培养基更换为无糖培养基,置于37℃培养箱(95%N2-5%CO2)中培养12 h,随后将培养基更换为正常培养基,37℃培养箱(95%空气-5%CO2)中继续培养24 h,制备氧糖剥夺-复氧复糖损伤模型。DHA组于氧糖剥夺前12 h时加入25μmol/L DHA孵育。采用MTT法测定细胞活力,采用化学比色法测定乳酸脱氢酶(LDH)活性,采用免疫荧光染色法计数活化小胶质细胞,采用Western blot法检测小胶质细胞活化标志物iba-1表达水平,采用ELISA法检测培养基TNF-α、IL-6、IL-4和IL-10的浓度。结果与C组比较,OGD/R组和DHA+OGD/R组细胞活力降低,LDH活性升高,活化小胶质细胞计数增多,iba-1表达上调,OGD/R组TNF-α和IL-6浓度升高,DHA+OGD/R组TNF-α、IL-6、IL-4和IL-10浓度升高(P<0.05);与OGD/R组比较,DHA组细胞活力升高,LDH活性降低,活化小胶质细胞计数减少,iba-1表达下调,TNF-α和IL-6浓度降低,IL-4和IL-10浓度升高(P<0.05)。结论DHA减轻氧糖剥夺-复氧复糖损伤的机制可能与抑制小胶质细胞活化,减轻炎症反应有关。Objective To evaluate the effect of docosahexaenoic acid(DHA)on microglial activation during oxygen-glucose deprivation and restoration(OGD/R)injury.Methods N9 microglia were inoculated in 96-well culture plates at a density of 104 cells/well for 3-5 days and divided into 3 groups(n=18 each)using a random number table method:control group(group C),group OGD/R and DHA+OGD/R group.The cells were cultured for 24 h in an incubator at 37℃(95%air-5%CO2)in group C.In OGD/R and DHA groups,the culture medium was replaced by glucose-free culture medium,the cells were cultured for 12 h in an incubator at 37℃(95%N2-5%CO2),and then cells were returned to the normal culture medium and cultured for 24 h in an incubator at 37℃(95%air-5%CO2)to establish the OGD/R injury model.The cells in group DHA were incubated with 25μmol/L DHA at 12 h before OGD/R.The cell viability was detected using the methyl thiazolyl tetrazolium assay,the activity of lactate dehydrogenase(LDH)was measured by using chemical colorimetric method,activated microglia were counted by immunofluorescence staining,the expression of microglia activation marker iba-1 was detected by Western blot,and the concentrations of tumor necrosis factor-α(TNF-α),interleukin 6(IL-6)and IL-4 and IL-10 in culture medium were determined using enzyme-linked immunosorbent assay.Results Compared with the group C,the cell viability was significantly decreased,the LDH activity was increased,the number of activated microglia was increased,and the expression of iba-1 was up-regulated in OGD/R and DHA+OGD/R groups,the concentrations of TNF-αand IL-6 were significantly increased in group OGD/R,and the concentrations of TNF-α,IL-6,IL-4 and IL-10 were significantly increased in group DHA+OGD/R(P<0.05).Compared with group OGD/R,the cell viability was significantly increased,the LDH activity was decreased,the number of activated microglia was decreased,the expression of iba-1 was down-regulated,TNF-αand IL-6 concentrations were decreased,and IL-4 and IL-10 concentrations were incr

关 键 词：二十二碳六烯酸类 再灌注损伤 小神经胶质细胞 

分 类 号：R28[医药卫生—中药学]