13-甲基十四烷酸对大鼠脑皮层星形胶质细胞氧反常的影响  

Effects of 13-MTD on cerebral ischemia and oxygen paradox in cultured astrocytes

作　　者：何宏星 翁绳美[2] 胡潇[3] 林艳婷 余涓[3] HE Hongxing;WENG Shengmei;HU Xiao;LIN Yanting;YU Juan(Laboratory Animal Center,Fujian Medical University,Fuzhou 350108,China;School of Pharmacy,Fujian Medical University,Fuzhou 350108;Department of Physiology and Pathophysiology,Fujian Medical University,Fuzhou 350108)

机构地区：[1]福建医科大学实验动物中心,福州350108 [2]福建医科大学药学院,福州350108 [3]福建医科大学基础医学院生理学与病理生理学系,福州350108

出　　处：《中国实验动物学报》2018年第4期454-460,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：福建省科技厅计划项目资助基金(No.2010Y0027);福建医科大学教授基金(No.JS12001)~~

摘　　要：目的研究13-甲基十四烷酸(13-methyltetradecanoic Acid,13-MTD)对大鼠脑皮质星形胶质细胞氧反常的保护作用。方法传代培养新生SD乳鼠大脑皮质星形胶质原代细胞,以氧糖剥夺/再复氧糖(OGD/R)方法复制氧反常模型,OGD 10 h/R 24 h,于再复氧糖即刻分别给予13-MTD 20,40,80μg/m L(M20,M40,M80)干预,倒置显微镜动态观察星形胶质细胞形态,细胞免疫化学鉴定角质纤维酸性蛋白(glial fibrillary acidic protein,GFAP),MTT法检测线粒体活性,免疫组化法检测星形胶质细胞的水通道蛋白4(aquaporin 4,AQP4)蛋白表达。结果OGD 10 h/R 24 h损伤后,体外培养的SD乳鼠脑皮质星形胶质细胞出现明显损伤,线粒体活性显著下降(P<0.01),星形胶质细胞膜AQP4蛋白表达量明显增加(P<0.01);与模型组比较,13-MTD 20,40,80μg/m L可减少损伤,使线粒体活性上升、AQP4蛋白表达减少,以80μg/m L效果最好(P<0.01)。结论 13-MTD可通过降低AQP4的表达,提高线粒体活性,减轻细胞水肿,进而保护氧反常诱导的星形胶质细胞损伤。Objective The aim of this work was to investigate the protective effects of 13-methyltetradecanoic acid（ 13-MTD） on focal cerebral ischemia in rats and oxygen paradox in cultured astrocytes in vitro. Methods Almost pure astrocytes were obtained after primary culture and subculture from cerebral cortex of neonatal rats,and then the OGD 10 h/R 24 h cell model was generated by treatment with oxygen-glucose deprivation for 10 h/reperfusion for 24 h. 13-MTD 20,40,80 μg/m L was administered immediately after reperfusion. Microscopy was used to observe the changes of cell morphology dynamically. MTT assay was employed to detect mitochondrial activity,and immunohistochemistry was done to determine the expressions of GFAP and AQP4. Results After oxygen paradox,damages of the cerebral cortical astrocytes of neonatal rats were observed,the mitochondrial activity was decreased significantly（ P〈 0. 01）,and AQP4 protein expression was increased significantly（ P〈 0. 01）. Compared with the vehicle control,13-MTD 20,40,80 μg/m L improved the above parameters（ P〈 0. 01）,with optimal effect being obtained at 80 μg/m L. Conclusions 13-MTD can down-regulated AQP4 protein expression,improve the mitochondrial activity,decrease cell edema,and protected the astrocytes damage from oxygen paradox.

关 键 词：13-甲基十四烷酸 脑缺血损伤 氧反常 星形胶质细胞 

分 类 号：Q95-33[生物学—动物学]