论文部分内容阅读
构建巨噬细胞验证模型,比较研究二氢槲皮素(Dihydroquercetin,taxifolin,TF)和二氢杨梅素(Dihydromyricetin,DMY)体外抗炎活性。以1 mg/L的脂多糖(LPS)诱导RAW264.7巨噬细胞建立了体外炎症模型;通过细胞毒性试验(MTT)法检测细胞存活率;Griess试剂法测定细胞上清液中NO(一氧化氮)释放量;ELISA法检测上清液中细胞因子IL-1β(白细胞介素-1β)、IL-6(白细胞介素-6)、TNF-α(肿瘤坏死因子α)和PGE2(前列腺素E2)的含量变化;通过RT-PCR法检测IL-1β、IL-6、TNF-α基因表达情况。MTT试验结果表明,二氢槲皮素和二氢杨梅素各剂量组没有表现出对巨噬细胞RAW264.7的细胞毒性。同时,二氢槲皮素高中低剂量组和二氢杨梅素高中低剂量组均能显著抑制脂多糖诱导细胞对NO、PGE2的释放(P<0.01),且均表现出剂量依赖性;二氢槲皮素高中低剂量组和二氢杨梅素高中低剂量组均能有效降低IL-1β、IL-6、TNF-α基因的表达,同时能够降低脂多糖诱导细胞产生细胞因子IL-1β、IL-6、TNF-α的含量。二氢槲皮素和二氢杨梅素均具有良好的体外抗炎作用,且二氢槲皮素的抗炎作用较好。
The model of macrophages was established to study the anti-inflammatory activity of Dihydroquercetin (TF) and Dihydromyricetin (DMY) in vitro. RAW264.7 macrophages were induced by lipopolysaccharide (LPS) at a concentration of 1 mg / L, and the cell viability was determined by cytotoxicity assay (MTT). The NO (monosodium glutamate The levels of IL-1β, IL-6, TNF-α and PGE2 in the supernatant were measured by ELISA. E2). The gene expression of IL-1β, IL-6 and TNF-α were detected by RT-PCR. MTT test results showed that dihydroquercetin and dihydromyricetin dose groups did not show macrophage RAW264.7 cytotoxicity. At the same time, dihydroquercetin high and low dose groups and dihydromyricetin high and low dose groups could significantly inhibit the release of NO and PGE2 by lipopolysaccharide-induced cells (P <0.01), and both showed a dose-dependent; dihydro High and low doses of quercetin group and high, medium and low doses of dihydromyricetin group were able to effectively reduce the expression of IL-1β, IL-6 and TNF-α gene, at the same time can reduce the production of lipopolysaccharide-induced cytokines IL-1β, IL -6, TNF-α content. Dihydroquercetin and dihydromyricetin have good anti-inflammatory effects in vitro, and dihydroquercetin better anti-inflammatory effect.