同型半胱氨酸诱导内皮细胞炎症损伤及金雀异黄酮的保护作用机制探讨

Effects of genistein on homocysteine-induced endothelial dysfunction and inflammatory response and its molecular mechanisms

目的:探讨金雀异黄酮(genistein,GEN)对同型半胱氨酸(homocysteine,Hcy)诱导的内皮细胞毒性和炎症的抑制作用及其分子机制。方法:采用MTT法检测细胞活性,倒置显微镜观察细胞形态变化,酶联免疫吸附法(ELISA)检测相关炎症分子白细胞介素-6(IL-6)和细胞间黏附分子-1(ICAM-1)。结果:单独加5.0 mmol/L的Hcy组活力最差,与对照组比较差异有统计学意义(P<0.01)。显微镜下见对照组细胞呈单层鹅卵石样紧密排列,细胞状态良好,而Hcy 5.0 mmol/L组细胞排列杂乱,凋亡增多,状态也最差。ELISA结果发现,与对照组比较,5.0 mmol/L的Hcy能显著增强IL-6及ICAM-1表达(P<0.01)。然而GEN(10、50、100μmol/L)能够增强细胞活力,显著抑制由Hcy介导的IL-6及ICAM-1表达的增强,大大改善细胞状态,尤其是100μmol/L的GEN(P<0.01)。结论:GEN对Hcy介导的内皮细胞毒性及炎症反应有明显的抑制作用。

Objective： To investigate the effects of genistein on homocysteine-mediated endothelial cell toxicity and inflammatory response and the underlying mechanisms of such effects. Methods： MTT assay was used to detect cell viability, changes in the morphology of cells were observed under microscope; ELISA assay was used to examine the levels of IL-6 and ICAM-1. Results：The viability of cells treated with 5.0 mmol/L of Hcy was lower than that of other groups, and the difference was statistics significance compared with the control group（ P 〈 0.01 ）. ECV-304 ceils were identified by their typical cobblestone morphology when viewed by light microscope in control group, the state of cells was also better, but the arrange of cells became disorderly, congregate with 5.0 mmol/L of Hcy in test groups,and the state was the worst. ELISA showed that homocysteine（5.0 mmoL/L）significantly increased the expression of IL-6 and ICAM-1 ,compared with the control group（ P 〈 0.01 ）. However,pretreatment with genistein （10,50,100 μmol/ L） reversed the decreased cell viability, improved the state of cells in a large extent,and suppressed an increase of expressions of IL-6 and ICAM-1 of cells induced by homcysteine markedly, especially the 100 μmol/L of genistein（ P 〈 0.01 ）. Conclusions： Genistein inhibits homocysteine-induced endothelial dysfunction and inflammatory response obviously.