摘要
目的:探讨基质细胞衍生因子1α(SDF-1α)对过氧化氢(H2O2)损伤人脑胶质瘤细胞U87的保护作用及机制。方法:双抗体夹心酶联免疫吸附试验(ELISA)检测胶质瘤细胞U87自分泌SDF-1α;细胞增殖实验研究外源SDF-1α对U87细胞增殖的影响;SDF-1α作用U87 12小时后,0.7 mM H2O2处理6小时,流式细胞术检测细胞凋亡率;蛋白质免疫印记实验(western blot)检测SDF-1α对U87细胞中蛋白激酶B(Akt)和细胞外信号调节激酶1/2(ERK1/2)磷酸化的影响。结果:胶质瘤细胞U87自身几乎不分泌SDF-1α,24小时内外源性SDF-1α对U87细胞增殖无明显影响;H2O2损伤后,SDF-1α预处理组细胞存活率高于对照组,凋亡率和死亡率低于对照组,差异具有统计学意义;Western blot显示SDF-1α处理能够诱导U87细胞Akt和ERK1/2的快速磷酸化。结论:SDF-1α能够提高H2O2损伤的U87细胞存活率,降低凋亡率和死亡率,其机制可能与磷脂酰肌醇3激酶(PI3K)-Akt和丝裂原活化蛋白激酶(MAPK)-ERK1/2通路的激活有关。
Objective：To investigate the protective effect and mechanism of stromal cell derived factor-1α（SDF-1α） on human glioma cell line U87 injuried by hydrogen peroxide（H2O2）.Methods：Enzyme-linked Immunosorbent Assay was used to detect the SDF-1α produced by U87 cells.MTT Assay was performed to evaluate the effect of proliferation on cells induced by human recombinant SDF-1α.Then cells were divided into three groups：control group,H2O2（0.7 mM） treated group,SDF-1α＋H2O2 treated group（100 ng/ml and 0.7 mM respectively）.Apoptotic cells were measured by flow cytometry.Western Blot was used to detect the expression of phosphorylated protein kinase B（pAkt） and phosphorylated extracellular signal regulating kinase（pERK1/2）.Results：The human glioma cells treated by H2O2 exhibited apoptosis in the presence of SDF-1 compared with the cells treated with H2O2 alone.100ng/ml SDF-1α treatement could lead to Akt and ERK activation in U87.Conclusion：SDF-1α could protect U87 cells from H2O2-induced apoptosis,the mechanism may be related to the activation of phosphatidylinositol 3-kinase（PI3K）-Akt and mitogen-activated protein kinase（MAPK）-ERK1/2 signalling pathway.
出处
《现代生物医学进展》
CAS
2011年第12期2266-2268,2272,共4页
Progress in Modern Biomedicine
基金
江苏省普通高校研究生科研创新计划项目(CX09B_264Z)
国家自然科学基金资助项目(编号:81072329)
