目的 研究大黄素对脂多糖（LPS）诱导人脐静脉血管内皮细胞（HUVEC）氧化损伤的保护作用及机制。方法 运用CCK-8细胞活力检测法筛选LPS体外诱导HUVEC细胞氧化损伤模型浓度及大黄素给药浓度。取HUVEC细胞，分为对照组、模型组（1 μg/L LPS）、吡咯烷二硫代氨基甲酸铵（PDTC，阳性药，10 μmol/L）组和大黄素低、高剂量（40、80 μmol/L）组，置于37℃、5% CO₂培养箱中培养24 h。ELISA法测定各组细胞上清液中一氧化氮（NO）、丙二醛（MDA）、活性氧（ROS）、肿瘤坏死因子-α（TNF-α）的含量；采用Western Blotting法检测各组细胞中Toll样受体（TLR4）、核因子κB（NF-κB p65）、TNF-α蛋白的表达。结果 LPS浓度为1 μg/L，作用24、48 h，HUVEC细胞存活率分别为64.8%、51.2%； 40、80 μmol/L大黄素作用24、48 h对HUVEC细胞存活率无显著影响，选择1 μg/L作为LPS造模浓度，40、80 μmol/L作用24 h作为大黄素给药条件。与对照组比较，模型组HUVEC细胞增殖活力显著下降，NO、MDA、ROS、TNF-α含量显著升高，TLR4、NF-κB p65、TNF-α蛋白表达升高（P<0.01）；与模型组比较，40、80 μmol/L大黄素给药后HUVEC细胞生存率显著升高，NO、MDA、ROS、TNF-α含量显著降低，TLR4、NF-κB p65、TNF-α蛋白表达显著降低（P<0.05、0.01）。结论 大黄素对LPS诱导的HUVEC细胞氧化损伤具有显著保护作用，其作用机制可能与调控TLR4/NF-κB通路、抑制炎症有关。

Objective To study the protective effect and mechanism of emodin (EM) on oxidative damage of human umbilical vein endothelial cells (HUVEC) induced by lipopolysaccharide (LPS). Methods CCK-8 cell viability test was used to screen the concentration of LPS induced oxidative damage model and emodin. HUVEC cells were divided into control group, model group (1 μg/L LPS), pyrrolidine dithiocarbamate (PDTC, positive drug, 10 μmol/L) group and emodin low and high dose (40, 80 μmol/L) group, and cultured in 37℃ and 5% CO₂ incubator for 24 h. ELISA was used to determine the contents of nitric oxide (NO), malondialdehyde (MDA), reactive oxygen species (ROS) and tumor necrosis factor (TNF-α) in the supernatant of each group and the western blotting was used to detect the expression of Toll-like receptor (TLR4), nuclear factor-κB (NF-κB p65), TNF-α protein expression in each group of HUVEC cells. Results Compared with control group, the proliferative activity of HUVEC cells in LPSinduced model group decreased significantly, the contents of NO, MDA, ROS and TNF-α increased significantly, and the expressions of TLR4, NF-kappa B p65, TNF-alpha and IL-6 increased significantly (P<0.01). After different concentrations of EM administration, the proliferation activity of HUVEC cells increased, the contents of NO, MDA, ROS and TNF-a decreased significantly, and the expressions of TLR4, NF-kappa B p65, TNF-a decreased significantly (P<0.05 and 0.01). Conclusion EM has a protective effect on LPS-induced oxidative damage in HUVEC endothelial cells, the mechanism may be related to the regulation of TLR4/NF-κB pathway, inhibit inflammation.

R285.5

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