知识与财富的链接
【目的】研究虾青素(AST)对脂多糖(LPS)通过TLR4/My D88/NF-κB信号通路诱导的IPEC-J2细胞炎症的影响。【方法】采用MTT法确定虾青素和LPS对IPEC-J2细胞和转染IPEC-J2细胞的最佳处理浓度和处理时间,在处理后采用实时荧光定量PCR检测IPEC-J2细胞和转染IPEC-J2细胞炎症因子NF-κB、TNF-α、IL-6和IL~(-1)β的m RNA相对表达量,采用ELISA检测上述炎症因子的分泌量。【结果】当处理3 h,虾青素浓度达到150μmol·L~(-1)时,IPEC-J2细胞和转染IPEC-J2细胞活力达到峰值;当处理6 h,LPS质量浓度达到100 ng·m L~(-1)时,IPEC-J2细胞和转染IPEC-J2细胞活力达到峰值。与对照组相比,LPS组IPEC-J2细胞NF-κB、TNF-α、IL-6、IL~(-1)β的m RNA相对表达量和分泌量显著升高(P0.05);与LPS组相比,AST+LPS组NF-κB、TNF-α、IL-6、IL~(-1)β在IPEC-J2细胞中的m RNA相对表达量和分泌量显著降低(P0.05),而在转染IPEC-J2细胞中的炎症因子m RNA相对表达量和分泌量均无显著变化(P0.05)。【结论】虾青素可以抑制细胞炎症反应,且对细胞的保护作用与TLR4/My D88/NF-κB信号通路相关。
【Objective】 To investigate the effect of astaxanthin (AST) on IPEC-J2 cell inflammation induced by lipopolysaccharide (LPS) via TLR4/MyD88/NF-κB signaling pathway.【Method】 MTT assay was performed to determine the optimal time and concentrations of astaxanthin and LPS for treating IPEC-J2 cells and transfected IPEC-J2 cells. Fluorescent quantitative RT-PCR was performed to determine the relative mRNA expressions of inflammatory factors including NF-κB, TNF-α, IL-6 and IL-1β in IPEC-J2 cells and transfected IPEC-J2 cells stimulated by LPS. ELISA assays were carried out to determine the secretion amounts of these inflammatory factors.【Result】 The vitality of IPEC-J2 cells and transfected IPEC-J2 cells reached the peak when treated with 150 μmol·L–1 astaxanthin for 3 h or 100 ng·mL–1 lipopolysaccharide for 6 h. Compared with control group, the relative mRNA expressions and secretions of NF-κB, TNF-α, IL-6 and IL-1β in IPEC-J2 cells significantly increased in LPS treatment group (P<0.05). Compared with LPS treatment group, the relative mRNA expressions and secretions of NF-κB, TNF-α, IL-6 and IL-1β in LPS+AST group were significantly lower in IPEC-J2 cells (P<0.05), but did not differ significantly in transfected IPEC-J2 cells (P>0.05).【Conclusion】 Astaxanthin can inhibit cell inflammation, and its protective effect on cells is related to TLR4/MyD88/NF-κB signaling pathway.
