高氧对星形胶质细胞的损伤及抗氧化剂对其保护作用  

The Damage of High Oxygen to Astrocytes and the Protective Effect of Antioxidants

作　　者：杨晖 张玲霞 叶雨露 董志章[1] 陈婕灵[1] 刘丹[1] 吴秋萍[1] 郑穗联[1] YANG Hui;ZHANG Lingxia;YE Yulu;DONG Zhizhang;CHEN Jieling;LIU Dan;WU Qiuping;ZHENG Suilian(The 2^nd Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院,温州325000

出　　处：《中国细胞生物学学报》2020年第6期945-953,共9页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金项目(批准号:LY15H120004)资助的课题。

摘　　要：该文旨在探究高氧环境对星形胶质细胞的损伤情况及抗氧化剂N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)对其的保护作用。该文将星形胶质细胞随机分为常氧组、高氧组(6 h、12 h、24 h)、常氧NAC组和高氧NAC组(6 h、12 h、24 h)。在高氧环境下培养不同时间后,检测星形胶质细胞的活性、细胞内线粒体膜电位的变化、细胞内的总活性氧(ROS)和线粒体ROS的改变,用免疫荧光染色法及Western blot检测细胞VEGF及VEGFR的表达情况。结果显示,高氧环境培养后,星形胶质细胞的活性显著下降(P<0.05)。细胞内总ROS水平及线粒体ROS水平较常氧组明显增加。星形胶质细胞在高氧环境下VEGF与VEGFR的表达下调。加入抗氧化剂NAC后,星形胶质细胞的细胞活性在高氧6 h和高氧12 h组增强(P<0.01),而在高氧24 h组减弱(P<0.05)。加入抗氧化剂后6 h组与12 h组星形胶质细胞内的ROS较未加抗氧化剂组明显减少,但对24 h高氧处理的星形胶质细胞内ROS无明显影响。加入抗氧化剂后高氧处理的星形胶质细胞的VEGF与VEGFR的表达量也有所增加,但在高氧24 h组,星形胶质细胞的VEGF和VEGFR表达仅少量增加。结果证实,星形胶质细胞在高氧条件下产生损伤,且与高氧持续时间相关,而使用NAC能够在一定程度上挽救短期内高氧对星形胶质细胞的损伤。以上结果提示,对高氧刺激后星形胶质细胞的抗氧化保护可能可以作为ROP治疗的一种方法。This study aimed to explore the damage of hyperoxia on astrocytes and the protective effect of antioxidants(N-acetyl-L-cysteine,NAC).Astrocytes were randomly divided into normoxic group,hyperoxic group(6 h,12 h,24 h),normoxic NAC group and hyperoxic NAC group(6 h,12 h,24 h).After different periods of exposure to hyperoxia,the activity of astrocytes,the changes of mitochondrial membrane potential in cells,the changes of total intracellular ROS(reactive oxygen species),mitochondrial ROS in cells,the expressions of VEGF and VEGFR were detected.The activity of astrocytes decreased significantly after hyperoxic culturing(P<0.05).The total intracellular ROS and mitochondrial ROS significantly increased compared with normoxic group.The expression of VEGF and VEGFR decreased in astrocytes in hyperoxic environment.After the using of the antioxidant NAC,the activity of astrocytes increased at 6 h and 12 h hyperoxia groups(P<0.01),but decreased at 24 h hyperoxia group(P<0.05).The ROS of astrocytes in the 6 h hyperoxic NAC group and the 12 h hyperoxic NAC group significantly reduced compared with those in the non-antioxidant group,but no significant changes of the ROS were observed in 24 h hyperoxic NAC group.VEGF and VEGFR levels also increased in hyperoxic astrocytes at 6 h and 12 h after the addition of the antioxidant.Astrocytes are damaged under high oxygen conditions and it is related to the duration of high oxygen.However,the use of NAC can save the damage of astrocytes caused by high oxygen in the short term to some extent,suggesting that the antioxidant protection of astrocytes after high oxygen stimulation may be a new therapy of ROP.

关 键 词：高氧 星形胶质细胞 抗氧化剂 血管内皮生长因子 早产儿视网膜病变 

分 类 号：R722.6[医药卫生—儿科] R774.1[医药卫生—临床医学]