摘要
目的:研究载脂蛋白A-I模拟肽L-4F是否减轻过氧化氢(hydrogen peroxide,H2O2)诱导的小鼠骨髓来源内皮祖细胞(endothelial progenitor cells,EPCs)氧化应激损伤及其机制。方法:通过密度梯度离心法分离小鼠骨髓单个核细胞,条件培养基EGM-2MV诱导分化培养EPCs。对培养EPCs先以PI3K/AKT抑制剂LY294002(30μmol/L)干预2 h后加入L-4F(75 mg/L)4 h,再加入100μmol/L H2O224 h。MTT法检测细胞活力,试剂盒检测各实验组培养基超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)含量,以评价细胞氧化与抗氧化平衡及脂质过氧化程度;流式细胞术Annexin V/PI双染法检测细胞凋亡;免疫印迹法检测p-AKT蛋白水平。结果:L-4F预处理显著抑制了H2O2诱导的细胞活力降低、SOD活性下降、MDA含量增加及细胞凋亡。H2O2下调p-AKT的蛋白水平,L-4F呈浓度依赖性地上调p-AKT的蛋白水平。LY294002抑制了L-4F减轻EPCs损伤的作用。结论:载脂蛋白A-I模拟肽L-4F部分通过PI3K/AKT通路减轻H2O2诱导的EPCs氧化应激损伤。
AIM： To investigate the protective effect and underlying mechanism of apolipoprotein A-I mimetic peptide L-4F on mouse bone marrow-derived endothelial progenitor cells（ EPCs） injured by hydrogen peroxide（ H2O2）.METHODS： The bone marrow-derived mononuclear cells of C57 mice were isolated by density gradient centrifugation and cultured in EGM-2MV medium. EPCs were pretreated with a PI3 K inhibitor LY294002（ 30 μmol / L） for 2 h,and then incubated with L-4F（ 75 mg / L） for 4 h,followed by the treatment with 100 μmol / L H2O2 for 24 h. MTT assay and flow cytometry with Annexin V / PI staining were used to detect cell viability and apoptosis. The extracellular superoxide dismutase（ SOD） andmalondialdehyde（ MDA） levels were measured to characterize the balance between oxidation and antioxidation,and lipid peroxidation. The protein level of phosphorylated AKT（ p-AKT） was analyzed by Western blot. RESULTS： L-4F restored H2O2-induced decrease in EPCs cell viability and apoptosis,and inhibited H2O2-induced decrease in SOD activity and increase in MDA level in culture medium. In addition,H2O2 inactivated p-AKT,which was significantly restored by L-4F. However,LY294002 inhibited the restoration of L-4F on EPCs impaired by H2O2. CONCLUSION： Apolipoprotein A-I mimetic peptide L-4F significantly reduced H2O2-induced mouse bone marrow-derived EPC injury partially through stimulating the PI3 K / AKT signaling pathway.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第1期1-7,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30971098)
山东省自然科学基金资助项目(No.Z2008C03
No.ZR2012HL18)
山东省教育厅科技计划(No.J14LK03)
关键词
动脉粥样硬化
载脂蛋白A-I模拟肽
内皮祖细胞
过氧化氢
氧化应激
Atherosclerosis
Apolipoprotein A-I mimetic peptide
Endothelial progenitor cells
Hydrogen per oxide
Oxidative stress
