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目的:研究蛇葡萄素F(AMPF)对脂多糖(LPS)致炎模型细胞的抗炎作用及其机制。方法:以RAW264.7细胞为模型细胞,采用CCK-8法测定不同质量浓度(100、40、20、10、5 μg/mL)的AMPF处理不同时间(24、48、72 h)后的细胞活性。设正常对照组、模型组和AMPF高、中、低剂量组(40、20、10 μg/mL),分别未加药物处理或加入相应质量浓度的AMPF溶液处理后培养24 h[除正常对照组外,其余各组均在培养1 h时加入LPS(1 μg/mL)致炎造模]。测定各组细胞中白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)和一氧化氮(NO)的含量,以及环氧合酶2(COX-2)和一氧化氮合酶(iNOS)的蛋白表达水平。结果:当培养时间为24 h时,AMPF各质量浓度组(100、40、20、10、5 μg/mL)细胞存活率均高于90%,AMPF未对细胞产生明显毒性。与模型组比较,AMPF高、中、低剂量组细胞中IL-1β的含量和iNOS的蛋白表达水平均显著降低,高剂量组细胞中IL-6、TNF-α、NO的含量均显著降低,高、中剂量组细胞中COX-2的蛋白表达水平均显著降低,差异均有统计学意义(P<0.05或P<0.01),并呈现一定的剂量依赖性。结论:AMPF对LPS诱导的RAW264.7细胞炎症具有显著的抑制作用,其机制可能与AMPF调控COX-2和iNOS的表达,进而抑制NO的释放,降低IL-1β、IL-6和TNF-α的含量有关。
OBJECTIVE: To study the anti-inflammatory effect of ampelopsin F (AMPF) on lipopolysaccharide (LPS)-induced inflammatory model cells and its mechanism. METHODS: Using RAW264.7 cell as model cell, CCK-8 assay was used to determine the activity of cell after treated for different duration (24, 48, 72 h) with different concentration (100, 40, 20, 10, 5 μg/mL) of AMPF. Then the cells were divided into control group, model group and AMPF high-dose, medium-dose and low-dose groups (40, 20, 10 μg/mL). They were cultured without or with relevant AMPF solution for 24 h [Except for normal control group, other groups received LPS (1 μg/mL) to induce inflammatory model after cultured for 1 h]. The contents of IL-1β, IL-6, TNF-α and NO, the protein expression of COX-2 and iNOS were detected in cell culture supernatant. RESULTS: After cultured for 24 h, survival rates of cells in AMPF groups (100, 40, 20, 10, 5 μg/mL) were all higher than 90%, showing no obvious cytotoxicity of AMPF. Compared with model group, the content of IL-1β and the protein expression of iNOS were decreased significantly in AMPF high-dose, medium-dose and low-dose groups; the contents of IL-6, TNF-α and NO were decreased significantly in AMPF high-dose group; the protein expression of COX-2 were decreased significantly in AMPF high-dose and medium-dose groups, with statistical significance (P<0.05 or P<0.01), in dose-dependent manner. CONCLUSIONS: AMPF shows significant anti-inflammatory effect on LPS-induced RAW264.7 cell inflammation, the mechanism of which may be associated with regulating the expression of COX-2 and iNOS, inhibiting the release of NO and decreasing the contents of IL-1β, IL-6, TNF-α and NO.
蛇葡萄素FRAW264.7细胞脂多糖炎症炎症因子
Ampelopsin FRAW264.7 cellslipopolysaccharideInflammationInflammatory factor
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