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目的:比较当归超临界总提取物及其馏分之间抗炎能力的差异,探讨是否存在作用更强的组分。方法:将当归超临界总提取物采用分子蒸馏技术在不同温度下分离出6个馏分,使用四甲基偶氮唑盐(MTT)法检测不同浓度药物组对小鼠巨噬细胞RAW264.7的细胞毒性。以脂多糖(LPS)刺激小鼠巨噬细胞RAW264.7建立炎症模型,亚硝酸盐(Griess)法检测炎症因子一氧化氮(NO)的含量、酶联免疫吸附法(ELISA)检测前列腺素(PGE_2)的释放量。实时荧光定量PCR法(qPCR)检测环氧化酶-2(COX-2)、诱导性一氧化氮合酶(iNOS)mRNA的表达。结果:各药物组均有抗炎作用,且馏分4的抗炎能力最强。结论:当归酯类成分之间按一定比例协同可发挥更好的抗炎作用。
OBJECTIVE: To compare the difference of anti-inflammatory abilities among the extracts of Angelica sinensis supercritical fluid and their fractions, and to explore whether there is a more effective component. Methods: Six fractions of Angelica sinensis supercritical extracts were separated by molecular distillation at different temperatures. MTT assay was used to detect the effect of different concentrations of drugs on RAW264.7 Cytotoxicity. Lipopolysaccharide (LPS) was used to stimulate mouse macrophage RAW264.7 to establish the model of inflammation. Nitric oxide (Griess) method was used to detect the content of nitric oxide (NO), and the content of prostaglandin PGE_2) release. Real-time quantitative PCR (qPCR) was used to detect the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) mRNA. Results: Each drug group had anti-inflammatory effect, and fraction 4 had the strongest anti-inflammatory ability. Conclusion: Angelica ester components synergistic with a certain proportion can play a better anti-inflammatory effect.