泛素对人脐静脉内皮细胞和巨噬细胞的影响

发布时间：2019-05-12 19:21

【摘要】：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(0.01μg/m L组、0.1μg/m L组、1μg/m L组和10μg/m L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0.1μg/m L泛素干预10μg/m L 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50μg/m L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0.1μg/m L和1μg/m L时明显增高,而在0.01μg/m L和10μg/m L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。0.1μg/m L泛素干预10μg/m L 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。
[Abstract]:Objective to compare and analyze the effects and differences of ubiquitin on (HUVECs) and macrophages in normal human umbilical vein endothelial cells (umbilical vein endothelial cells), and to evaluate whether ubiquitin has protective effect on HUVECs damage induced by lipopolysaccharide (LPS). Methods the morphological changes of HUVECs cells treated with different concentrations of ubiquitin were observed under microscope. HUVECs and human macrophages cultured in vitro were divided into blank control group and four ubiquitin intervention groups (0.01 渭 g / mL group, 0.1 渭 g / mL group, 1 渭 g / mL group and 10 渭 g / mL group). The levels of tumor necrosis factor 伪 (TNF- 伪) and vascular cell adhesion molecule 1 (VCAM-1) were measured by ELISA assay 24 hours after intervention. The levels of nuclear factor kappa B (NF- kappa B) protein were measured by, Western blot assay. HUVECs,ELISA assay induced by 0.1 渭 g / mL ubiquitin in 10 渭 g / m L LPS was used to determine the levels of TNF- 伪 and VCAM-1 in the cells of ubiquitin LPS group and LPS group at 16 h and 24 h. Results with the increase of the concentration of ubiquitin, the degree of HUVECs damage was gradually aggravated. When the concentration of ubiquitin reached 50 渭 g / mL, the cells began to become balloon-like and died a lot. With the increase of ubiquitin concentration, the levels of TNF- 伪 and VCAM-1 in HUVECs cells increased at first and then decreased, while the levels of TNF- 伪 and VCAM-1 in human macrophages increased gradually. The levels of NF- 魏 B protein in HUVECs and human macrophages increased significantly when the concentration of ubiquitin was 0.1 渭 g / mL and 1 渭 g / mL, but not at 0.01 渭 g / mL and 10 渭 g / mL, and even decreased at 0.01 渭 g / mL and 10 渭 g / mL. The decrease trend of HUVECs was more obvious. The levels of TNF- 伪 and VCAM-1 in LPS group and ubiquitin LPS group induced by 0.1 渭 g / mL ubiquitin at 10 渭 g / m L LPS were similar at each time point, but there was no significant difference. Conclusion low concentration of ubiquitin has obvious damage effect on HUVECs, while human macrophages need higher concentration of ubiquitin. Ubiquitin has no obvious protective effect on HUVECs damage induced by LPS in vitro.
【作者单位】：南京明基医院重症医学科;南京明基医院胸腔医学中心;
【基金】：南京医科大学科技发展基金面上项目(编号:2011NJMU076)
【分类号】：R392

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