川芎嗪对内皮祖细胞氧化应激损伤的保护作用研究

 目的观察川芎嗪(TMP)对过氧化氢(H₂O₂)诱导的内皮祖细胞(EPCs)氧化应激损伤的保护作用及机制。方法通过密度梯度离心分离脐静脉血单个核细胞,利用血管内皮生长因子(VEGF)诱导纯化方法培养EPCs。CCK-8细胞计数试剂盒、流式细胞仪、激光共聚焦显微镜检测TMP对细胞氧化应激损伤的保护作用及机制探讨。结果200mg·L^–1 TMP抑制内皮祖细胞增殖功能;但是25mg·L^-1 TMP显著减少H₂O₂致细胞凋亡和细胞生长停滞在G0/G1期,而L-单甲基精氨酸(L-NMMA)抵消TMP的保护作用。结论TMP对氧化应激损伤诱导细胞凋亡和生长停滞具有明显的保护作用,eNOS受体拮抗剂L-NMMA能抵消TMP这种保护作用,说明TMP对EPCs抗氧化应激损伤的保护作用可能是通过eNOS途径。

Protective Effect of Tetramethylpyrazine on Oxidative Stress Injured Endothelial Progenitor Cells in Vitro

OBJECTIVE To study the protective effect and mechanisms of tetramethylpyrazine (TMP) on oxidative stress injured endothelial progenitor cells by hydrogen peroxide (H₂O₂). METHODS Total mononuclear cells were isolated from cord blood by ficoll density gradient centrifugation, and cultured with vascular endothelial growth factor (VEGF) in vitro. The cells cultured for 7 d were divided into control, H₂O₂ model, TMP, TMP+H₂O₂ and L-NMMA+TMP+H₂O₂ groups. Detected protective effect and the relevant mechanisms of TMP on oxidative stress injured cells were measured by cck-8 cell count kit, flow cytometry, confocal Laser microscopy. RESULTS The high concentrations of 200 mg·L^-1 TMP inhibited the proliferation of endothelial progenitor cells,the low concentration of 25 mg·L^–1 TMP showed protective effect on EPCs oxidative stress. The results showed that TMP+H₂O₂ group significantly reduced the hydrogen peroxide-induced cells apoptosis and cells detention in G0/G1 stage, compared with H₂O₂ group(P<0.05). The nitricoxide synthase (eNOs) inhibitor NG-monomethyl L-arginine(L-NMMA ) counteracted the protective effect of TMP. There were no significantly different in cells apoptosis and cells development, compared with H₂O₂ group. CONCLUSION TMP protects EPCs from oxidative injured stress, including cells apoptosis and cells growth by hydrogen peroxide. The benefit of TMP to EPCs is offseted by L-NMMA. It means eNOs may play a role in this anti-oxidative stress injury effect.