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目的研究肥大细胞(或肥大细胞裂解上清液)、平滑肌细胞与氧化低密度脂蛋白(oxLDL)共孵育时,肥大细胞对平滑肌细胞源性泡沫细胞形成的影响。方法用oxLDL处理大鼠腹腔肥大细胞后,采用荧光分光光度仪检测组胺释放率,通过倒置显微镜、甲苯胺蓝染色、透射电镜观察肥大细胞脱颗粒状态。将肥大细胞(或肥大细胞裂解上清液)、平滑肌细胞与60mg/LoxLDL共育48h,以油红O染色观测细胞内中性脂质,高效液相色谱法检测细胞内胆固醇和胆固醇酯含量。结果肥大细胞被oxLDL激活脱颗粒,肥大细胞脱颗粒度与oxLDL呈剂量依赖性关系,以60mg/LoxLDL处理肥大细胞时,肥大细胞脱颗粒度达71.70%±3.02%;甲苯胺蓝染色和透射电镜均显示,oxLDL处理后肥大细胞明显脱颗粒。在60mg/LoxLDL存在时,以肥大细胞(或肥大细胞裂解上清液)处理平滑肌细胞48h后,油红O染色显示,处理组胞浆内脂滴明显多于未处理组;高效液相色谱分析结果显示,肥大细胞裂解上清液处理组细胞内总胆固醇和胆固醇酯含量明显高于未处理组,总胆固醇增高了0.57倍,胆固醇酯增高了0.83倍。结论oxLDL以剂量依赖性方式诱导肥大细胞脱颗粒;肥大细胞脱颗粒后促进oxLDL诱导的平滑肌细胞源性泡沫细胞形成。
Objective To investigate the effect of mast cells on the formation of SMC-derived foam cells when mast cells (or mast cell lysis supernatant), smooth muscle cells and oxidized low density lipoprotein (oxLDL) were co-incubated. Methods After treatment of peritoneal mast cells with oxLDL, the release rate of histamine was detected by fluorescence spectrophotometer. The degranulation status of mast cells was observed by inverted microscope, toluidine blue staining and transmission electron microscopy. The mast cells (or mast cell lysis supernatant) and smooth muscle cells were incubated with 60mg / L oxLDL for 48h. The intracellular lipids were stained with Oil Red O and the content of cholesterol and cholesterol esters in the cells were detected by HPLC. Results Mast cells were degranulated by oxLDL. Mast cell degranulation was in a dose-dependent manner with oxLDL. When treated with 60 mg / L oxLDL, mast cells degranulation reached 71.70% ± 3.02%. Toluidine blue staining and transmission electron microscopy All show that mast cells were significantly degranulated after oxLDL treatment. Smooth muscle cells were treated with mast cells (or mast cell lysis supernatant) for 48 h in the presence of 60 mg / L oxLDL. Oil red O staining showed that there were more lipid droplets in the cytoplasm of the treated group than those in the untreated group. High performance liquid chromatography The results showed that the content of total cholesterol and cholesterol ester in the cells treated with mast cell lysis supernatant was significantly higher than that of the untreated group, the total cholesterol increased by 0.57 times and the cholesterol ester increased by 0.83 times. Conclusion OxLDL induces degranulation of mast cells in a dose-dependent manner; mast cells degranulation promotes oxLDL-induced smooth muscle cell-derived foam cell formation.