天麻素对氧糖剥夺模型中小鼠少突胶质前体细胞的保护作用  

Protective effects of Gastrodin on oligodendrocyte precursor cells in vitro model of oxygen glucose deprivation

作　　者：郭艳吉 郭家智 钟莲梅[1] 陶俊 姚玥伊 艾明达 陆地 艾青龙[1] Guo Yanji;Guo Jiazhi;Zhong Lianmei;Tao Jun;Yao Yueyi;Ai Mingda;Lu Di;Ai Qinglong(Kunming Medical University,The First Affiliated Hospital,Department of Internal Neurology,Kunming 650032;Kunming Medical University,Biomedical Engineering Research Center,Kunming 650500,China)

机构地区：[1]昆明医科大学第一附属医院神经内科,昆明650032 [2]昆明医科大学生物医学工程研究中心,昆明650500

出　　处：《神经解剖学杂志》2019年第6期599-605,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金资助项目(No.81460210,81460350);昆明医科大学2018年研究生创新基金(2018S099)

摘　　要：目的:探讨天麻素对氧糖剥夺(OGD)模型中小鼠少突胶质前体细胞(OPCs)的保护作用。方法:采用来自于C57BL/6小鼠的原代少突胶质前体细胞进行OGD模型实验。首先分为对照组和OGD不同时间处理模型组,用CCK-8试剂盒检测小鼠OPCs的细胞活力变化。用细胞凋亡-Hoechst染色试剂盒检测各组在不同时间凋亡细胞数量的变化。最后选择细胞活性开始明显下降以及凋亡细胞开始明显增多的时间点(18 h)作为天麻素对抗OGD模型中小鼠OPCs凋亡增多的实验研究时间点,采用Western Blot和免疫荧光细胞化学染色检测18 h时cleaved-caspase 3、Bax和Bcl-2的表达变化。结果:小鼠OPCs的细胞生长曲线结果表明,OGD模型组的细胞在12 h前细胞活力应激性升高,6 h时达到最高,12 h后细胞活力逐渐下降,18 h时细胞活力明显低于对照组(P <0. 05)。此外,18 h时,OGD模型组的凋亡细胞较对照组增多,天麻素治疗组能逆转凋亡细胞数量的升高,并且能下调OGD模型组中小鼠OPCs中cleaved-caspase3和Bax蛋白的表达量(P <0. 01),上调抑制Bcl-2蛋白及Bcl-2/Bax的比值(P <0. 01)。结论:OGD处理18 h时能引起小鼠OPCs细胞活力下降以及凋亡增多,天麻素能够抑制小鼠OPCs的凋亡。Objective: To investigate the protective effects of gastrodin on mouse oligodendrocyte precursor cells(OPCs) in the model of oxygen glucose deprivation(OGD). Methods: Experiments were divided into control group and OGD groups conducted within varied time points using primary OPCs from C57BL/6 mouse. We adopt CCK-8 to detect the cell viability of mouse OPCs,and apoptosis-Hoechst staining kit to observe the changes of apoptotic cell number at different time in control group,OGD model group and groups pretreated with gastrodin(10 μmol/L). Finally,choose the time point(18 h) when OPCs cell activity began to decrease while apoptotic cells began to increase significantly as the experimental time point of gastrodin antagonizing OPCs apoptosis increasing in OGD model. The diversification of apoptosis-related protein were detected by Western Blot and immunofluorescence staining for cleaved-caspase 3 and Box,Bcl-2 immunoreactivities at 18 h. Results: The results of OPCs growth curve showed that the cell viability of OGD group increased before 12 h,reached the highest level at 6 h,decreased gradually after 12 h,and was significantly lower than that of blank control group at 18 h.(P < 0. 05). In addition,the number of OPCs apoptotic cells in OGD group was higher than that in control group at 18 h. The group pretreated with gastrodin can not only reverse the increase of apoptotic cells,but also down-regulate the expression of cleaved-caspase3 and Bax protein of mouse OPCs(P < 0. 01),whilst up-regulate the protein of Bcl-2 and the ratio of Bcl-2/Bax under the condition of OGD(P <0. 01). Conclusion: The viability of mouse OPCs declines,and the number of apoptotic cells increases simultaneously under the condition of OGD at 18 h,furthermore,gastrodin can inhibit the apoptosis of mouse OPCs.

关 键 词：天麻素 OPCs OGD 凋亡 氧糖剥夺模型 小鼠 

分 类 号：R28[医药卫生—中药学]