目的 研究大黃素對脂多糖（LPS）誘導(dǎo)人臍靜脈血管內(nèi)皮細胞（HUVEC）氧化損傷的保護作用及機制。方法 運用CCK-8細胞活力檢測法篩選LPS體外誘導(dǎo)HUVEC細胞氧化損傷模型濃度及大黃素給藥濃度。取HUVEC細胞，分為對照組、模型組（1 μg/L LPS）、吡咯烷二硫代氨基甲酸銨（PDTC，陽性藥，10 μmol/L）組和大黃素低、高劑量（40、80 μmol/L）組，置于37℃、5% CO₂培養(yǎng)箱中培養(yǎng)24 h。ELISA法測定各組細胞上清液中一氧化氮（NO）、丙二醛（MDA）、活性氧（ROS）、腫瘤壞死因子-α（TNF-α）的含量；采用Western Blotting法檢測各組細胞中Toll樣受體（TLR4）、核因子κB（NF-κB p65）、TNF-α蛋白的表達。結(jié)果 LPS濃度為1 μg/L，作用24、48 h，HUVEC細胞存活率分別為64.8%、51.2%； 40、80 μmol/L大黃素作用24、48 h對HUVEC細胞存活率無顯著影響，選擇1 μg/L作為LPS造模濃度，40、80 μmol/L作用24 h作為大黃素給藥條件。與對照組比較，模型組HUVEC細胞增殖活力顯著下降，NO、MDA、ROS、TNF-α含量顯著升高，TLR4、NF-κB p65、TNF-α蛋白表達升高（P<0.01）；與模型組比較，40、80 μmol/L大黃素給藥后HUVEC細胞生存率顯著升高，NO、MDA、ROS、TNF-α含量顯著降低，TLR4、NF-κB p65、TNF-α蛋白表達顯著降低（P<0.05、0.01）。結(jié)論 大黃素對LPS誘導(dǎo)的HUVEC細胞氧化損傷具有顯著保護作用，其作用機制可能與調(diào)控TLR4/NF-κB通路、抑制炎癥有關(guān)。

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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