獐牙菜苦苷的抗炎活性及其对NF-κB通路相关因子p65和IKK-α表达的影响

Anti-inflammatory Activity of Swertiamarin and Its Effect on the Expression of NF-κB Pathway-related Factors p65 and IKK-α

关键词:  獐牙菜苦苷  巨噬细胞  抗炎活性  核转录因子-κB

Key words:  Swertiamarin  macrophage  anti-inflammatory activity  NF-κB

摘要:

目的 观察獐牙菜苦苷（Swertiamarin）的抗炎活性以及对核转录因子-κB（NF-κB）通路的调控作用。方法 采用脂多糖（LPS）诱导小鼠巨噬细胞RAW264.7构建体外炎症模型，CCK-8法观察獐牙菜苦苷对细胞活性的影响，ELISA法检测其对炎症相关因子肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）表达的影响，RT-PCR法测定其对NF-κB通路相关因子p65和IKK-α的mRNA表达的影响。结果 100 μg·mL^-1獐牙菜苦苷能降低细胞存活率，3.125~50 μg·mL^-1浓度对细胞存活率无影响；25~50 μg·mL^-1獐牙菜苦苷能降低炎症相关因子TNF-α及IL-6的释放；12.5~50 μg·mL^-1獐牙菜苦苷能抑制p65的mRNA表达，50 μg·mL^-1獐牙菜苦苷能抑制IKK-α的mRNA表达。结论 獐牙菜苦苷对LPS诱导的炎症模型中TNF-α、IL-6的生成具有抑制作用，其机制可能与抑制NF-κB通路相关因子p65、IKK-α的表达有关。

Abstract:

OBJECTIVE To observe the anti-inflammatory activity of Swertiamarin and its regulation effect on the nuclear transcription factor-κB(NF-κB) pathway. METHODS Lipopolysaccharid(LPS) induced murine macrophage raw 264.7 cell was used to establish an inflammatory model in vitro. CCK-8 method was used to observe the effect of Swertiamarin on cell activity. ELISA was used to detect its effect on the expression of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6). RT-PCR was used to detect its effect on mRNA expression of NF-κB pathway related factors p65 and Ikk-α. RESULTS The 100 μg·mL^-1 Swertiamarin could decrease the cell survival rate, and 3.125-50 μg·mL^-1 had no effect on the cell survival rate. The 25-50 μg·mL^-1 Swertiamarin could reduce the release of TNF-α and IL-6. It was found that 12.5-50 μg·mL^-1 Swertiamarin could inhibit the mRNA expression of p65, and the mRNA expression of Ikk-α could be inhibited by 50 μg·mL^-1 Swertiamarin. CONCLUSION Swertiamarin has inhibitory effect on TNF-α and IL-6 production in LPS-induced inflammatory model, and its mechanism may be related to the inhibition of the expression of p65 and IKK-α, which are related factors of NF-κB pathway.