该文探讨草苁蓉多糖(Boschnikia rossica polysaccharides,BRPS)对脂多糖(lipopolysaccharide,LPS)诱导的小鼠J774A.1巨噬细胞炎症反应的抑制作用。将小鼠J774A.1巨噬细胞随机分为正常组、模型组、BRPS组(剂量分别为25、50、100 mg/L),用细胞增殖毒性试剂盒检测J774A.1巨噬细胞存活率,酶联免疫吸附法检测细胞培养液中的肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平。用免疫印迹法测定环氧合酶-2(COX-2)、Toll样受体4(TLR4)、白介素-1受体相关激酶4(IRAK4)、髓样分化因子88(MyD88)以及核转录因子-κB(NF-κB)蛋白表达情况。结果表明,LPS在100~2 000 μg/L浓度范围时对J774A.1巨噬细胞存活率无影响,但能够上调小鼠J774A.1巨噬细胞炎症因子TNF-α和IL-6的分泌,此结果提示J774A.1细胞炎症模型构建成功。BRPS对LPS诱导的J774A.1细胞存活率无影响;但是能抑制LPS诱导的J774A.1巨噬细胞TNF-α、IL-6以及IL-1β的分泌;同时,与模型组比较,BRPS组J774A.1巨噬细胞中COX-2、TLR4、IRAK4、MyD88以及核NF-κB蛋白表达水平降低,此结果提示BRPS对LPS诱导的小鼠J774A.1细胞炎症反应起到抑制作用,其抗炎作用可能与其抑制TLR4 /NF-κB信号通路有关。
The inhibitory effect of Boschniakia rossica polysaccharides (BRPS) on the inflammatory response of J774A.1 macrophages induced by lipopolysaccharide (LPS) was investigated. The inflammation model of macrophage was established by stimulating J774A.1 macrophage with LPS. The J774A.1 cell was randomly divided into the normal group, model group and BRPS groups (the concentrations of BRPS were 25, 50 and 100 mg/L, respectively). Cell viability was detected by cell counting kit-8, and the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by the enzyme-linked immunosorbent assay. The protein expressions of cyclooxygenase-2 (COX-2), toll-like receptor 4 (TLR4), interleukin-1 receptor-associated kinase 4 (IRAK4), myeloid differentiation factor 88 (MyD88) and nuclear factor-κB (NF-кB) were determined with the western blot method. The results showed that LPS had no obvious effect on the viabilities of J774A.1 macrophages in the concentration range of 100-2 000 μg/L. However, the secretion of TNF-α and IL-6 were increased which indicating the establishment of macrophage inflammation model. BRPS could reduce LPS-induced secretion of IL-1β, IL-6 and TNF-α into the culture medium of J774A.1 cell. At the same time, BRPS down-regulated the protein expressions of COX-2, TLR4, IRAK4, MyD88 and nuclear NF-кB induced by LPS. It is suggested that BRPS has an inhibitory effect on LPS-induced inflammation in mouse J774A.1 cells, which may be related to the inhibition of the TLR4/NF-κB pathway.
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