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-  2015 

泛素对人脐静脉内皮细胞和巨噬细胞的影响

DOI: doi:10.7507/1671-6205.2015044

Keywords: 泛素, 人脐静脉内皮细胞, 人巨噬细胞, 脂多糖

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Abstract:

目的对比分析泛素对正常状态下的人脐静脉内皮细胞(HUVECs)和巨噬细胞的影响及其差别,并评价泛素是否对脂多糖(LPS)诱导的HUVECs损伤具有保护作用。 方法在显微镜下观察不同浓度泛素干预后HUVECs细胞形态学的变化。将体外培养的HUVECs和人巨噬细胞分为空白对照组和4个泛素干预组(0.01 μg/mL组、0.1 μg/mL组、1 μg/mL组和10 μg/mL组),ELISA法测定干预后24 h各组细胞上清肿瘤坏死因子α(TNF-α)和血管细胞粘附分子1(VCAM-1)水平,Western blot法测定细胞核因子κB (NF-κB)蛋白水平。用0.1 μg/mL泛素干预10 μg/mL LPS诱导的HUVECs,ELISA法测定8、16和24 h泛素+LPS组和LPS组细胞上清TNF-α和VCAM-1水平。 结果随着泛素干预浓度的增加,HUVECs损伤的程度逐步加重,当泛素浓度达到50 μg/mL时,细胞开始呈气球样变并大量死亡。随着泛素干预浓度的升高,HUVECs细胞上清TNF-α和VCAM-1水平呈先升高后降低的变化趋势,而人巨噬细胞的TNF-α和VCAM-1水平则是呈逐步升高的趋势;HUVECs和人巨噬细胞NF-κB蛋白水平在泛素浓度为0.1 μg/mL和1 μg/mL时明显增高,而在0.01 μg/mL和10 μg/mL时增高不明显,甚至有减低的趋势,其中HUVECs减低的趋势更为明显。0.1 μg/mL泛素干预10 μg/mL LPS诱导的HUVECs,各时间点LPS组和泛素+LPS组的TNF-α和VCAM-1水平相似,差异无统计学意义。 结论较低浓度的泛素对HUVECs即有明显的损伤作用,而人巨噬细胞则需要较高的泛素浓度才出现损伤。泛素对体外培养的LPS诱导的HUVECs损伤不具有明显的保护作用

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