虾青素对脂多糖诱导的RAW264.7细胞炎症反应的影响及机制

    Effect of astaxanthin on inflammatory response of RAW264.7 cells induced by lipopolysaccharide and its mechanism

    • 摘要:
      目的  研究虾青素(AST)对脂多糖(LPS)诱导的RAW264.7细胞炎症反应的影响及作用机制,为将虾青素应用于炎症治疗奠定理论基础。
      方法  采用不同浓度梯度的脂多糖及虾青素对RAW264.7细胞进行不同时间段的处理,通过MTT法确定最佳处理浓度和时间,对细胞进行最佳处理后,用ELISA法、荧光定量PCR技术和Western blot法分别检测细胞炎症因子的分泌量、mRNA相对表达量和蛋白相对表达量。
      结果  100 μmol/L虾青素和2 μg/mL脂多糖处理3 h的RAW264.7细胞活力处于峰值。与对照组相比,脂多糖组RAW264.7细胞中TNF-α、IL-6和Caspase-1的分泌量分别降低了12.83%、9.66%和20.80%(P<0.05),脂多糖对于TLR4/MyD88/NF-кB通路相关蛋白表达有促进作用,其中,TLR4和NF-кB p65蛋白相对表达量分别提高了195.40%和226.95%(P<0.05);与LPS组相比,AST+LPS组中虾青素对炎性因子的分泌及mRNA表达有抑制作用,TLR4、MyD88和NF-кB p65蛋白相对表达量降低了54.99%、45.70%和28.20%(P<0.05)。
      结论  虾青素预保护能抑制TLR4/MyD88/NF-кB通路相关蛋白的表达,进而缓解脂多糖刺激RAW264.7细胞产生的炎症反应。

       

      Abstract:
      Objective  To study the effect of astaxanthin (AST) on the inflammatory response of RAW264.7 cells induced by lipopolysaccharide (LPS) and the mechanism, and provide a theoretical basis for using AST in inflammation therapy.
      Method  Different concentrations of LPS and AST were used to treat RAW264.7 cells for different time. The optimal treatment concentration and time were determined by MTT method. After applying the optimal treatment, the secretion, mRNA relative expression and protein relative expression of inflammatory factors were detected by ELISA, fluorescence quantitative PCR and Western blot method respectively.
      Result  When treated with 100 μmol/L AST and 2 μg/mL LPS for 3 h, the viability of RAW264.7 cells was at the peak. Compared with the control group, the secretion of TNF-α, IL-6 and Caspase-1 in RAW264.7 cells of LPS group reduced by 12.83%, 9.66% and 20.80% respectively(P<0.05). LPS promoted the expression of TLR4/MyD88/NF-κB pathway-related proteins with relative expression of TLR4 and NF-кB p65 proteins enhanced by 195.40% and 226.95% respectively(P<0.05). Compared with LPS group, AST had inhibitory effects on the secretion and mRNA expression of inflammatory factors in AST+LPS group, and the relative expression of TLR4, MyD88 and NF-кB p65 proteins reduced by 54.99%, 45.70% and 28.20% respectively (P<0.05).
      Conclusion  AST pre-protection can inhibit the expression of TLR4/MyD88/NF-κB pathway-related proteins, thereby alleviate the inflammatory response in RAW264.7 cells induced by LPS.

       

    /

    返回文章
    返回