巨噬细胞增强H2O2抑制人血管内皮细胞和兔血管平滑肌细胞的增殖

摘要

目的 探讨H2O2处理对巨噬细胞分泌因子的影响,及对血管内皮细胞(VEC)及平滑肌细胞(VSMC)增殖的影响.方法 实验分组:空白对照组,0.5 mmol/L H2O2组,巨噬细胞条件培养基组,巨噬细胞+0.5 mmol/L H2O2条件培养基组.CCK-8测定VEC和VSMC的增殖情况.ELISA检测白介素石(IL-6)、白介素-10(IL-10)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α),以及血管内皮细胞生长因子(VEGF).Western blot检测增殖核抗原(PCNA)、细胞周期蛋白cyclinD1的表达,以及血管内皮细胞内皮源性一氧化氮合酶(eNOS)的表达.结果 VEC中其他3个处理组的增殖水平均低于对照组(P＜0.05),而在VSMC中,巨噬细胞组的增殖水平高于空白对照组(P＜0.05).在VEC和VSMC中,巨噬细胞+0.5 mmol/L H2O2组的增殖水平均高于单独0.5 mmol/L H2O2处理组(P＜0.05),且低于单独巨噬细胞处理组(P＜0.05).巨噬细胞分泌的IL-6、IL-10、MCP-1随时间和H2O2的浓度增加而增高(P＜0.05).VEC分泌的VEGF在1.0 mmol/L H2O2浓度时有明显增高(P＜0.05).结论 氧化应激处理可影响巨噬细胞炎性因子的分泌,进而影响血管内皮细胞和平滑肌细胞的增殖.%Objective Investigate the changes of inflammatory factors secreted by macrophages exposed to oxidative stress,and their effect on the proliferation of vascular endothelial cells and smooth muscle cells.Methods The groups described as followings:control,macrophage,0.5 mmol/L H2O2,macrophage + 0.5 mmol/L H2O2.The growth curves of VECs and VSMCs under different conditions were measured by CCK-8.The IL-6,IL-10,MCP-1,TNF-αt and VEGF were detected by ELISA.The level of PCNA,cyclinD1 and eNOS were analysised by Western blot.Results The proliferation of control group was higher than the others in VEC (P ＜ 0.05),while in VSMC,the proliferation of control group was lower than that of macrophage group (P ＜ 0.05).In both VEC and VSMC,macrophages + 0.5 mmol/L H2O2 conditioned medium groups were higher than 0.5 mmol/L H2O2 groups (P ＜0.05),while were lower than macrophage groups (P ＜0.05).The secretions of IL-6,IL-10,and MCP-1 in macrophages were increased with time and H2O2 concentration(P ＜0.05),while the titer of TNF-α didn't change significantly.Only the concentration of H2O2 reached 1.0 mmoL/L,and secretion of VEGF increased significantly (P ＜ 0.05).Conclusions The oxidative stress affect the secretions of inflammatory cytokines secreted by macrophages,and thus regulate proliferations of vascular endothelial cells and smooth muscle cells.