反油酸对血管内皮细胞功能及形态的影响

EFFECT OF ELAIDIC ACID ON MORPHOLOGY AND FUNCTION OF ENDOTHELIAL CELLS

目的通过建立血管内皮细胞(EC)与血管平滑肌细胞(SMC)的联合培养模型,探讨反油酸对内皮细胞的损伤作用。方法将与平滑肌细胞联合培养的内皮细胞用浓度为50μmol/L的油酸和反油酸分别作用48h。试剂盒测定乳酸脱氢酶(LDH)活力、一氧化氮(NO)含量;RT-PCR测定细胞粘附因子(VCAM-1、ICAM-1和E-slectin)基因表达量;激光共聚焦显微镜观察处理后的内皮细胞形态。结果反油酸促进LDH渗出,且紧密接触模型中内皮细胞LDH渗出率较旁分泌模型显著增加;反油酸抑制内皮细胞NO分泌,两种模型间无显著性差异;紧密接触模型中内皮细胞与旁分泌模型中的内皮细胞相比,VCAM-1、ICAM-1、E-slectin基因表达量均显著上升,且反油酸促粘附因子表达作用强于油酸;与平滑肌细胞紧密接触的内皮细胞出现形态不规则,反油酸使内皮细胞变形。结论与平滑肌联合培养可增加反油酸对内皮细胞的损伤。

Objective To investigate the effect of elaidic acid on endothelial cells （EC）. Methods Co-cultured endothelial cells and smooth muscle cells （SMC） models were used. The co-cultured cells were treated with 50 μmol/L oleic acid or 50 μmol/L elaidic acid for 48 hours, then lactic acid dehydrogenase （LDH）, and nitrogen oxide （NO） were measured spectrophotometrically. Gene expression of vascular cell adhesion molecule-1 （VCAM-1）, intercellular adhesion molecule-I （ICAM-1） and E-selectin were detected by RT-PCR respectively. Cell morphology of endothelial cells was observed by laser scanning confocal microscope. Results Elaidic acid increased LDH leakage, especially in EC/SMC （tightly contact） model. NO level was decreased with the addition of elaidic acid in both EC＋SMC （separated） or EC/SMC models. The expressions of VCAM-1, ICAM-1 and E-selectin of endothelial cells were significantly increased in EC/SMC model. Also, elaidic acid induced irregularity and transformation of endothelial cells membrane. Conclusion The injury on endothelial cells by elaidic acid is exacerbated when co-cultured with smooth vascular cells.