不同时间窗MACO模型大鼠脑脊液对星形胶质细胞活性的影响  

Effects of cerebrospinal fluid from rat MACO model with different time windows on activity of astrocytes

作　　者：张光茹[1] 宫圣 王界成[1] 李玉华[1] 王志华[1] 王学习[2] 

机构地区：[1]青海省中医院脑病科,青海西宁810000 [2]兰州大学基础医学院,甘肃兰州730000

出　　处：《兰州大学学报（医学版）》2016年第3期1-5,共5页Journal of Lanzhou University（Medical Sciences）

基　　金：国家自然科学基金项目(81360207);青海省(应用)基础研究项目(2012-Z-737);兰州市城关区科技发展项目(2015-3-2);兰州大学中央高校基本科研业务费专项资金项目(lzujbky-2015-156)

摘　　要：目的观察MCAO模型大鼠不同时间点脑脊液炎性反应及其对星形胶质细胞增殖和谷氨酸摄取能力的影响。方法制备MCAO模型大鼠,采集造模后30 min、6 h和24 h脑脊液,Elisa法检测脑脊液中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)水平;3种脑脊液干预体外培养星形胶质细胞,孵育6、24、48 h后MTT法检测星形胶质细胞增殖情况。根据星形胶质细胞增殖检测结果,选择6 h脑脊液继续干预星形胶质细胞,孵育6、24、48 h后进行乳酸脱氢酶(LDH)、星形胶质细胞谷氨酸摄取能力检测。结果 MCAO模型大鼠脑缺血30 min后,脑脊液中炎性因子IL-1β、IL-6水平未有变化,6 h后IL-1β、IL-6水平升高,24 h后IL-1β、IL-6水平仍然较高,但与6 h比较未进一步升高。MCAO模型大鼠30 min、6 h和24 h脑脊液干预星形胶质细胞24 h和48 h,星形胶质细胞增殖活性均降低,6 h和24 h脑脊液的抑制作用强于30 min脑脊液,但6 h脑脊液和24 h脑脊液二者的抑制作用无差异。MCAO模型大鼠6 h脑脊液干预星形胶质细胞,LDH水平升高,且干预时间越长,LDH水平越高。6 h脑脊液干预星形胶质细胞6 h和24 h,星形胶质细胞谷氨酸摄取能力增加,但干预48 h后其谷氨酸摄取能力降低。结论脑缺血6 h后,脑脊液中炎性因子IL-1β、IL-6水平升高,同时可造成星形胶质细胞损伤,抑制星形胶质细胞增殖,但对星形胶质细胞的谷氨酸摄取能力影响是先增强后抑制。Objective To observe inflammatory responses of cerebrospinal fluid to rat brain ischemia at different time and their influence on the proliferation and glutamate uptake ability of astrocytes. Methods Cerebrospinal fluid（CSF） was collected at 30 min, 6 h, 24 h after the rat middle cerebral artery occlusion（MCAO）model was established. ELISA method was used to detect the levels of inflammatory cytokines IL-1β, IL-6 in CSF. CSF collected from 3 time points was used to intervene astrocytes in vitro. After incubated for 6 h, 24 h,48 h, the proliferation of astrocytes was measured by MTT method. According to the astrocytes proliferation assay results, 6 h CSF was selected to incubate with astrocytes for 6 h, 24 h, 48 h, the lactic dehydogencse（LDH） level and glutamate uptake ability were detected. Results The levels of inflammatory cytokines IL-1β,IL-6 did not change in CSF 30 min after cerebral ischemia, however, the levels were increased 6 h after ischemia and maintained high level 24 h after ischemia, but there was no significant difference between this two time points. The proliferation of astrocytes were decreased after intervened by CSF from MCAO model rats at30 min, 6 h and 24 h, the inhibitory effect of CSF at 6 h and 24 h were stronger than that at 30 min, but no difference between 6 h and 24 h of CSF. Incubation of CSF collected at 6 h after ischemia with astrocytes, the level of LDH was increased, and the longer the intervention, the higher the LDH level. When CSF collected at6 h after ischemia intervening astrocytes for 6 h, 24 h, the glutamate uptake ability of astrocytes were increased, but decreased after intervention for 48 h. Conclusion The elevated inflammatory cytokines IL-1β and IL-6 in CSF after cerebral ischemia can cause injury of astrocytes, inhibit proliferation of astrocytes, but their impact on glutamate uptake ability of astrocytes was firstly enhanced and then inhibited.

关 键 词：缺血再灌注 星形胶质细胞 脑脊液 

分 类 号：R363[医药卫生—病理学]