SIRT1对激活型小胶质细胞BV-2毒性损伤PC12细胞的保护作用及机制研究  

Protection role of silent information regulator 1 in toxicity of activated BV-2 cells to PC12 ceils

作　　者：叶杰明[1] 刘振华[1] 谢惠芳[1] 魏继鹏[1] 陆伶俐[1] 黄燕君[1] 

机构地区：[1]南方医科大学珠江医院神经内科,广州510282

出　　处：《中华神经医学杂志》2013年第11期1112-1117,共6页Chinese Journal of Neuromedicine

摘　　要：目的探讨沉默信息调控因子1（SIRTl）在激活型小胶质细胞介导PCI2细胞损伤中所起的作用及相关机制。方法体外常规培养BV-2小胶质细胞和PCI2细胞，ELISA检测1μg／mL脂多糖（LPS）作用BV-2细胞6、12、24h后上清肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）的水平；MTT检测LPS与BV-2细胞共培养上清对PCI2细胞存活率的影响，同时设对照组、LPS组、单纯BV-2上清液组；MTT检测SIRTl激活剂藜芦醇（5、10、25、50、100μmol／L1、SIRTl抑制剂尼克酰胺（5、10、25、50mmol／L）对PCI2细胞存活率的影响，以筛选二者的干预浓度。实验分对照组、LPS＋BV-2细胞共培养上清液组、白藜芦醇干预组、尼克酰胺干预组，分别加入培养基、LPS＋BV-2细胞共培养上清液、50μmol／L白藜芦醇、25mmoFL尼克酰胺，培养18h后MTT检测4组细胞存活率，Westernblotting检测细胞内SIRT1、乙酰化p53的表达水平。结果LPS作用BV-2细胞6、12、24h后IL-6、TNF-α分泌量均依次增高，差异有统计学意义（P〈0．05）；MTT比色显示与对照组、LPS组、单纯BV-2上清液组比较，LPS＋BV-2细胞共培养上清组PCI2细胞存活率下降，差异有统计学意义（氏0．05）；100μmol／L白藜芦醇组、50μmol／L尼克酰胺组细胞存活率较对照组降低，差异有统计学意义（P〈0．05），因此选择50μmol／L白藜芦醇、25μmol／L尼克酰胺进行实验；与对照组比较，LPS＋BV-2细胞共培养上清组细胞存活率降低，SIRT1的表达下降，乙酰化p53的表达上升．差异有统计学意义（P〈0．05），与LPS与BV-2共培养上清组比较，白藜芦醇干预组细胞活性上升，SIRT1的表达水平较高，乙酰化p53的表达量较低，尼克酰胺干预组结果相反，细胞活性下降，SIRT1的表达较低，乙酰化p53的表达较高，差异有统计学意义（P〈0．05）。结论在LPS与小胶质细胞共培养上清损伤多巴胺（DA）能神经元�Objective To observe the effects of silent information regulator 1 （SIRT1） on toxicity of activated BV-2 to PC12 cells and the possible mechanisms. Methods BV-2 microglial cells and PC12 cells were routinely cultured in vitro; ELISA was used to measure to the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） stimulated by lipopolysaccharide （LPS, 1 μg/mL） in BV-2 cells. MTT assay was employed to identify the cell viability of PC12 cells injured by culture medium of activated BV-2 and determined the suitable concentrations of resveratrol （a potent SIRT1 activator, 5, 10, 25, 50 and 100 μmol/L） and nicotinamide （a known SIRT1 inhibitor, 5, 10, 25 and 50 mmol/L）. PC12 cells were divided into groups as follows： control group II, LPS＋BV-2 co-cultred group, resveratrol treatment group and nicotinamide treatment group （pretreated with resveratrol or nicotinamide for 2 h, and then subjected to culture medium of activated BV-2 cells, in the presence of resveratrol or sirtinol for 18 h）; the cell viability was measured by OD value in MTT assay, and the expressions of SIRT1 and acetyl-p53 were detected by Western blotting. Results TNF-α and IL-6 secretions increased gradually at 6, 12 and 24 h after LPS being induced BV-2, with significant difference between each two time points （P〈0.05）. PC12 cell viability decreased in the LPS＋BV-2 co-cultured group as compared with that in the control group I, LPS treatment group and BV-2 supernate group （P〈0.05）. The cell viability of cells in the 100 μmol/L resveratrol treatment group and 50 p, mol/L niacinamide treatment group decreased as compared with that in the control group Ⅱ （P〈0.05）, therefore, 50 μmol/L resveratrol and 25 μmol/L nicotinamide were chosen in the next experiment. As compared with those in the control group III, the cell viability and SIRT1 expression significantly decreased, and acetyl-p53expression significantly increased in the LPS＋BV-2 co-cultured group （P〈0.05）; as compared

关 键 词：沉默信息调控因子 帕金森病 BV-2细胞 大鼠肾上腺嗜铬细胞瘤细胞 P53 

分 类 号：R741[医药卫生—神经病学与精神病学]