[关键词]
[摘要]
探讨紫甘蓝总花色苷提取物对脂多糖(LPS,1 μg/mL)诱导RAW264.7细胞炎症反应的影响。以不同质量浓度(1 μg/mL、10 μg/mL、50 μg/mL)的紫甘蓝总花色苷提取物处理RAW264.7细胞后,试剂盒法分别检测一氧化氮(NO)和前列腺素E2(PGE2)的分泌水平。酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6和IL-8分泌水平。实时定量PCR(Quantitative real-time PCR,qRT-PCR)检测细胞中诱导型一氧化氮合酶(iNOS)、环氧合酶-2(COX-2)、TNF-α、IL-1β、IL-6和IL-8的mRNA表达水平。结果显示,紫甘蓝总花色苷提取物能有效下调iNOS与COX-2的mRNA表达,抑制NO和PGE2的释放。特别地,高浓度的紫甘蓝总花色苷提取物(50 μg/mL)能显著地抑制LPS所诱发的iNOS(65.80%)与COX-2(48.28%)的mRNA表达,降低NO(58.81%)和PGE2(46.26%)的释放水平。同时,紫甘蓝总花色苷提取物还能显著下调TNF-α、IL-1β、IL-6和IL-8的mRNA表达水平,并抑制TNF-α、IL-1β、IL-6和IL-8的分泌。结果表明,紫甘蓝总花色苷提取物具有较强的抗炎活性,能够抑制LPS诱导的RAW264.7细胞所发生的炎症反应。
[Key word]
[Abstract]
The effects of total anthocyanins from red cabbage (RCTA, 1 μg/mL-50 μg/mL) on lipopolysaccharide (LPS, 1 μg/mL) induced inflammatory reaction in RAW264.7 cells were investigated. The cellular levels of nitric oxide (NO) and prostaglandin E2 (PGE2) were determined by commercial kits. Tumor necrosis factor-α (TNF-α), interlukin (IL)-1β, IL-6 and IL-8 were determined by commercial enzyme linked immunosorbent assay (ELISA) kits according the manuscripts. The mRNA expressions of induced nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and inflammatory cytokines related TNF-α, IL-1β, IL-6 and IL-8 were determined by quantitative real time polymerase chain reaction (qRT-PCR) assay. The results indicated that, RCTA significantly reduced the LPS induced secretion of inflammatory mediators (NO and PGE2) and inflammatory cytokines (TNF-α、IL-1β、IL-6 and IL-8) in RAW264.7 cells. The high concentration of RCTA (50 μg/mL) was effectively reduced the mRNA expressions of iNOS (65.80%) and COX-2 (48.28%), and generations of (58.81%) and PGE2 (46.26%) in LPS treated RAW264.7 cells. In addition, RCTA treatment was able to reduce the mRNA expressions of iNOS, COX-2, TNF-α, IL-1β, IL-6 and IL-8, respectively. The results demonstrated that the RCTA exhibited a protective effect to reduce the secretion of inflammatory mediators and cytokines.
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[基金项目]
国家自然科学基金资助项目(81560530;81760589);广西自然科学基金资助项目(2016GXNSFCA380026);桂林医学院引进人才科研启动基金(04010150001);2018年广西教育厅广西高等学校千名中青年骨干教师培育计划资助(桂教人[2018]18号)