紫菜薹总花色苷对脂多糖诱发RAW264.7细胞炎症反应的影响

Effect of total anthocyanins from Brassica campestris L on lipopolysaccharide-induced inflammatory reaction in RAW264.7 cells

探讨紫菜薹总花色苷提取物对脂多糖(LPS)诱发RAW264.7细胞炎症反应的调控作用。MTT法测定紫菜薹总花色苷对细胞的细胞毒性效果。酶联法(Enzyme linked immunosorbent assay,ELISA)法检测细胞内一氧化氮(NO)、前列腺素E2(PGE2),肿瘤坏死因子-α(TNF-α)、白介素(Interlukin,IL)-1β、IL-6和IL-8的分泌水平。实时荧光定量PCR(Quantitative real time polymerase chain reaction,q RT-PCR)测定细胞内诱导型一氧化碳合酶(i NOS)、环氧合酶2(COX-2)及TNF-α、IL-1β、IL-6和IL-8等炎性细胞因子的m RNA转录水平。紫菜薹总花色苷提取物对细胞无明显的细胞毒性作用,能显著地抑制模型细胞中炎性介质(NO和PGE2)和炎性细胞因子(TNF-α、IL-1β、IL-6和IL-8)的分泌。同时,紫菜薹总花色苷还能显著抑制i NOS、COX-2以及TNF-α、IL-1β、IL-6和IL-8的mRNA转录。结果提示,紫菜薹总花色苷能通过抑制细胞炎性介质以及炎性细胞因子的活性来显著改善LPS所诱发RAW264.7细胞发生的炎症反应。

To investigate the effect of total anthocyanins from Brassica campestris(BCTA)on lipopolysaccharide(LPS)induced inflammatory reaction in RAW264.7 cells.Cell viability was determined by MTT assay.The cellular levels of nitric oxide(NO),prostaglandin E2(PGE2),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).BCTA showed non cytotoxic for RAW264.7 cells.BCTA also 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.In addition,BCTA treatment was able to reduce the mRNA expressions of iNOS,COX-2,TNF-α,IL-1β,IL-6 and IL-8,respectively.The results from this study suggested that the BCTA exhibited a protective effect to reduce the secretion of inflammatory mediators and cytokines,which to against LPS-induced inflammatory reaction in RAW264.7 cells.