脂质过氧化诱导培养的内皮细胞产生单核细胞趋化因子RANTES
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国家自然科学基金(项目编号39730220)资助


Lipid Peroxidation Induces Production of Monocyte Chemotactic Factor RANTES
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    摘要:

    为探讨脂质过氧化损伤是否诱导内皮细胞产生单核细胞趋化因子RANTES ,使培养的人脐静脉内皮细胞暴露于5 μmol/L联胺后收取其条件培养基。继之,将其放在截留分子质量分别为3.5kDa(外层)和10kDa(内层)的双层透析袋中,间层注入适量的0 .0 1mol/LPBS ,再将透析袋悬在装有PBS的锥形瓶内进行透析。透析完毕,取出间层液体,即为分子质量为3 .5~10kDa的条件培养基,其中含RANTES。用微孔滤膜法进行单核细胞趋化实验,并应用鼠抗人RANTES抗体,以证明条件培养基中RANTES的趋化活性。结果发现,经联胺刺激的条件培养基引起的单核细胞迁移距离(89.75±11.33μm)明显大于不经联胺刺激的条件培养基(77.30±11.5 3μm)和随机移动组(76 .18±10 .5 0 μm)。方差分析显示,组间差异有极显著性(F =47.2 0 ,P<0 .0 1)。加入鼠抗人RANTES抗体后,单核细胞迁移距离明显缩短(F =2 1.31,P<0 .0 1)。结果提示,从功能实验方面表明脂质过氧化损伤能诱导内皮细胞产生RANTES增强,并可能在动脉粥样硬化病变的发生过程中起一定作用。

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    Aim To understand whether lipid peroxidation injury to endothelial cells ( ECs ) induces the production of monocyte chemotactic factor RANTES. Methods After a four hour exposure of the cultured human umbilical vein ECs to 5 μmol/L diamide, the conditioned medium (CM) was collected. To obtain a conditioned medium containing RANTES (MW 8 kDa ), the above CM was put into the double layer dialysis molecular porous membrane tubings (Spectrum Lab. Inc. ) with the molecular weight cut off at 10 kDa ( inner layer ) and 3.5 kDa ( outer layer ) respectively. Appropriate amount of 0.01 mol/L PBS was injected into the interspace between the two membranes. The whole set of the membrane tubings was suspended in a pyramid flask containing PBS, and then the samples were dialysed against PBS. Finally, a conditioned medium with MW of 3.5~10 kDa was taken out from the interspace between the two membranes. The chemotactic activity of this conditioned medium for monocytes was tested by micropore filter method using a modified Boyden chamber. For antibody inhibition studies, the conditioned medium was incubated with anti-RANTES antibody and assayed again. Results The monocyte migration distance induced by diamide-CM group (89.75±11.33 μm) is markedly longer than that of the nondiamide-CM group(77.30±11.53 μm)and that of the random migration group(76.18±10.50 μm ). Analysis of variance showed that there was a significant difference between groups(F=47.20,P<0.01). After the addition of anti-RANTES antibody, monocyte chemotactic activity was markedly inhibited (F=21.31,P<0.01 ). Conclusion On the basis of functional assay it suggests that lipid peroxidation injury might induce ECs to produce increased RANTES and may play an role in the recruitment of monocytes into the intima in atherogenesis.

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赵霞,祝学卫,杨丽敏,邓仲端,朱大和.脂质过氧化诱导培养的内皮细胞产生单核细胞趋化因子RANTES[J].中国动脉硬化杂志,2001,9(4):281~284.

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  • 收稿日期:2000-12-27
  • 最后修改日期:2001-07-10
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