Annexin—A1模拟肽Ac2—26抑制小胶质细胞释放炎性介质的作用研究  

Effects of annexin-A1 mimetic peptide Ac2-26 on inflammatory factors released from microglia

作　　者：罗振钊[1] 刘璐[2] 马健[1] 王静[1] 胡绘[1] 孔曼[1] 王九菊[1] 施静[2] 卢忠心[1] Luo Zhenzhao Liu Lu Ma Jian Wang Jing Hu Hui Kong Man Wang Jiuju Shi Jing Lu Zhongxin(Department of Clinical Laboratory,Wuhan Central Hospital Department of Neurobiology , Tongji Medical College, Huazhong University of Science and Technology , Wuhan 430014, China)

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院检验科,430014 [2]华中科技大学同济医学院基础医学院神经生物学系,430014

出　　处：《中华老年医学杂志》2016年第12期1320-1323,共4页Chinese Journal of Geriatrics

基　　金：国家自然科学基金（31171029、31471015）;武汉市中心医院博士基金（YB15804）

摘　　要：目的观察Annexin—A1模拟肽Ac2—26对小胶质细胞炎性介质肿瘤坏死因子-α（TNF—α）、白介素-1β（IL-1β）、白介素-6（IL-6）和一氧化氮（NO）释放的影响及其对Toll样受体2（TLR2）的调控作用。方法根据处理不同将大鼠来源的原代小胶质细胞随机分为Aβ1-42组（给予Aβ1-42刺激组）、Aβ1-42＋对照乱序肽组（在给予Aβ1-42刺激的同时加入氨基酸乱序组处理）、Aβ1-42＋Ac2—26（在给予Aβ1-42刺激的同时加入Ac2—26处理）及空白对照组。经上述处理24h后，收集各组小胶质细胞。用CCK-8法检测细胞存活率，采用倒置相差显微镜观察细胞形态，收集各组细胞上清液采用酶联免疫吸附法（ELISA）测定TNF-α、IL-1β、IL-6和NO的浓度，采用Western blot法测定TLR2的蛋白水平。结果Aβ1-42和Ac2—26对小胶质细胞存活率没有影响，与空白对照组比较差异无统计学意义。Aβ1-42作用后，小胶质细胞形态发生明显变化，由静息的苎麻状变成激活的阿米巴状巨噬细胞样；Aβ1-42组、Aβ1-42＋对照乱序肽组、Aβ1-42＋Ac2—26及空白对照组上清TNF-α浓度分别为（125．17±7．13）ng／L（118．78±7．28）ng／L、（24．02±2．62）ng／L和（20．89±1．82）ng／L，IL-1β浓度分别为（117．61±8．73）ng／L（108．90±6．53）ng／L、（27．06±3．32）ng／L和（24．58±3．45）ng／L，IL-6浓度分别为（108．67±5．86）ng／L（102．67±4．85）ng／L（25．94±2．83）ng／L和（19．68±2．66）ng／L，Aβ1-42＋Ac2-26组上清液TNF-α、IL-1β和IL-6浓度明显低于Aβ1-42组、Aβ1-42＋对照乱序肽组（P〈0．05），与空白对照组比较差异无统计学意义。Aβ1-42＋Ac2—26组上清液NO浓度为（20．27±2．53）mmol／L，低于Aβ1-42组（75．68±6．14）mmol／L、Aβ1-42＋对照乱序肽组（69．25±4．50）mmol／L，与空白对照组（16．39±2．96）mmol／L比较差异无统计学意义。Western blObjective Objective To observe the effect of Annexin-Al mimetic peptide Ac2-26 on microglia released tumor necrosis factor-a（ TNF-a）, interleukin-1 b（ IL-1b）, interleukin-6 （ IL-6 ） and nitric oxide （NO）, and to explore its impact on expression of Toll Like Recepto〉2（TLR2）. Methods Rat primary microglial cells were randomly divided into 4 groups by different treatments ：Aβ1-42 group （ stimulated by Aβ1-42 ）,Aβ1-42 plus scramble control（co-stimulated by Aβ1-42 and scramble control）, Aβ1-42 plus Ac2-26 （costimulated by Aβ1-42and Ac2-26）and blank control group. After above treatments for 24 hour, the microglial cells were collected. Microglial cell viability was measured by CCK-8 assay kits. The inverted phase contrast microscope was applied to observe the morphological changes of microglia. The concentrations of TNF-a, IL-1β, IL-6 and NO in the supernatant were measured by commercial kits, and the level of TLR2 protein expression was detected by Western blot assay. Results The Aβ1-42 and Ac2-26 had no effect on the microglial cell viability. After activation with Aβ1-42, the microglia showed an obvious morphological change. The microglia cell soma showed obvious changes, from resting ramified state into the activated amoeboid macrophage-like appearance. The Aβ1-42 group, Aβ1-42 ＋ scramble control group, Aβ1-42＋ Ac2-26 group and blank control group showed that the supernatant levels of TNF-a were （125.17±7.13）, （118. 78±7.28）, （24. 02± 2. 62）, （20. 89 ±1.82） ng/L, respectively, the supernatant levels of IL-1β were （117. 61±8. 73）,（108. 90±6.53）, （27. 06±3.32）, （24. 58±3.45）ng/L,respectively,and the supernatant levelsof IL-6 were（108.6 7± 5.86）, （102.67±4.85）, （25.94 ±2.83）, （19.68 ±2.66） ng/L, respectively. The supernatant levels of TNF-a,IL-1β and IL-6 were significantly lower in the Aβ1-42＋Ac2-26 group than in Aβ1-42 group and Aβ1-42 ＋ scramble control groups, and had no significant difference c

关 键 词：阿尔茨海默病 小神经胶质细胞 Ac2—26 炎症 TOLL样受体2 

分 类 号：R749.16[医药卫生—神经病学与精神病学]