Annexin-A1模拟肽Ac2-26对大鼠星形胶质细胞活化的影响  

Effect of Annexin-A1 mimetic peptide Ac2-26 on activation of astrocytes in rats

作　　者：罗振钊[1] 魏礼清[1] 胡绘 孔曼[1] 王静[1] 谭哲琼 朱满 李星 施静[2] 卢忠心[1] Luo Zhenzhao;Wei Liqing;Hu Hui;Kong Man;Wang Jing;Tan Zheqiong;Zhu Man;Li Xing;Shi Jing;Lu Zhongxin(Department of Medical Laboratory,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China;Department of Neurobiology,The School of Basic Medical Science,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院检验科,430014 [2]华中科技大学同济医学院基础医学院神经生物学系,武汉430030

出　　处：《中华麻醉学杂志》2019年第8期948-952,共5页Chinese Journal of Anesthesiology

基　　金：武汉市卫计委青年项目(WX17Q10)。

摘　　要：目的评价Annexin-A1模拟肽Ac2-26对大鼠星形胶质细胞活化的影响。方法原代培养大鼠皮层组织星形胶质细胞,按随机排序的方法分为4组(n=24):对照组(C组)、LPS组、LPS+乱序肽对照组(LPS+Src组)和LPS+Ac2-26组。LPS组加入LPS终浓度1 mg/ml;LPS+Src组加入LPS终浓度1 mg/ml和乱序肽终浓度3.3 mmol/L;LPS+Ac2-26组加入LPS终浓度1 mg/ml和Ac2-26终浓度3.3 mmol/L。孵育24 h后采用CCK-8法测定细胞存活率,采用Transwell小室测定细胞迁移率,ELISA法测定上清液肿瘤坏死因子-α(TNF-α)、白介素1-β(IL-1β)、单核细胞趋化蛋白-1(MCP-1)和巨噬细胞炎性蛋白1α(MIP-1α)浓度,Western blot检测星形胶质细胞神经胶质酸性蛋白(GFAP)、细胞外信号调节激酶(ERK)、磷酸化ERK(p-ERK)、c-Jun N-末端激酶(JNK)、磷酸化JNK(p-JNK)、p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化p38MAPK(p-p38MAPK)的表达水平。计算p-ERK/ERK比值、p-JNK/JNK比值、p-p38MAPK/p38MAPK比值。结果与C组比较,LPS组GFAP表达上调,迁移率、上清液TNF-α、IL-1β、MCP-1和MIP-1α浓度、p-ERK/ERK比值、p-JNK/JNK比值和p-p38MAPK/p38MAPK比值升高(P<0.05),细胞存活率差异无统计学意义(P>0.05);与LPS组比较,LPS+Ac2-26组GFAP表达下调,迁移率、上清液TNF-α、IL-1β、MCP-1和MIP-1α浓度、p-JNK/JNK比值和p-p38MAPK/p38MAPK比值降低(P<0.05),LPS+Src组上述指标差异无统计学意义(P>0.05)。结论Ac2-26可抑制大鼠星形胶质细胞活化,产生抗炎作用。Objective To evaluate the effect of Annexin-A1 mimetic peptide Ac2-26 on activation of astrocytes in rats.Methods The primarily cultured astrocytes from the cortex of fetal Sprague-Dawley rats after 4 passages were divined into 4 groups(n=24 each)using a random number table method:control group(C group),LPS group,LPS+scramble peptide group(LPS+Src group)and LPS+Ac2-26 group.LPS was added to LPS group with the final concentration of 1 mg/ml.LPS at the final concentration of 1 mg/ml and scramble peptide at the final concentration of 3.3 mmol/L were added to LPS+Src group.LPS at the final concentration of 1 mg/ml and Ac2-26 at the final concentration of 3.3 mmol/L were added to LPS+Ac2-26 group.After 24-h incubation,the cell survival rate was measured by CCK-8 assay,the migration was determined by Transwell assay,the concentrations of tumor necrosis factor-alpha(TNF-α),interleukin-1beta(IL-1β),monocyte chemoattractant protein-1(MCP-1)and macrophage inflammatory protein-1a(MIP-1a)in the supernatant were measured(by enzyme-linked immunosorbent assay),and the expression of glial fibrillary acidic protein(GFAP),extracellular signal-regulated kinase(ERK),phosphorylated ERK(p-ERK),c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),p38 mitogen-activated protein kinase(p38MAPK),and phosphorylated p38MAPK(p-p38MAPK)in astrocytes was detected by Western blot.ResultsCompared with group C,the expression of GFAP was significantly up-regulated,and the cell mobility,concentrations of TNF-α,IL-1β,MCP-1 and MIP-1αin the supernatant,p-ERK/ERK ratio,p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were increased(P<0.05),and no significant change was found in the cell survival rate in group LPS(P>0.05).Compared with group LPS,the expression of GFAP was significantly down-regulated,and the cell mobility,concentrations of TNF-α,IL-1β,MCP-1 and MIP-1αin the supernatant,p-JNK/JNK ratio and p-p38MAPK/p38MAPK ratio were decreased in group LPS+Ac2-26(P<0.05),and no significant change was found in the parameters mentioned above in group

关 键 词：星形细胞 Ac2-26 

分 类 号：R28[医药卫生—中药学]