非酯化脂肪酸对体外培养犊牛原代肝细胞中炎性因子表达及其活性的影响

在建立犊牛原代肝细胞体外培养模型的基础上，通过培养液中添加不同浓度的非酯化脂肪酸（Nonesterifiedfatty acids，NEFAs），运用实时荧光定量PCR和ELISA方法，测定NEFAs对肝细胞中炎性因子TNFα、IL-6和IL-1β的mRNA表达及其活性的影响。结果显示，NEFAs作用肝细胞9h后，与对照组相比，高浓度NEFAs（1．8mmol／L）组肝细胞中TNFα、IL-1β的mRNA表达水平显著增加（P〈0．05），IL-6的mRNA表达水平在2．4retool／L时极显著增加（P〈0．01），低浓度NEFAs（0．6、1．2mmol／L）组肝细胞中TNFα、IL-1β和IL-6的mRNA表达水平无显著差异（P〉0．05）；并且TNFα、IL-1G的活性在NEFAs浓度达到1．8、2．4mmol／L显著高于对照组（P〈0．01），且呈剂量依赖性作用，IL-6的活性在在NEFAs浓度达到1．8mmol／L显著高于对照组（P〈0．01），在2．4mmol／L也高于对照组，但差异不显著。结果表明，高浓度NEFAs在-定程度上能引起或加剧奶牛肝细胞炎性反应，可能是导致产后奶牛肝功能炎性损伤主要因素。

Effects of NEFAs on expression and activity of inflammatory cytokines in calf hepatocytes cultured in vitro

This study aimed to investigate the effects of non-esterified fatty acid （NEFAs） on expression and activity of inflammatory cytokines in calf hepatocytes cultured in vitro. The total RNA was isolated from hepatocytes,which were treated with different concentrations of NEFAs after 9 h,and extraction cell supernatant was used for ELISA assay. The results showed that the mRNA levels of TNFα,IL-1β increased significantly （P〈0.05） in NEFAs （1.8 mmol/L）-treated,and the mRNA levels of IL-6 increased significantly （P〈0.01） in NEFAs （2.4 mmol/L ）- treated compared with the control group. In addition, the activity of TNFα, IL-1β were significantly higher than that in control group in NEFAs （1.8 mmol/L and 2.4 mmol/L）-treated,and the activity of IL-6 were significantly higher in NEFAs （1.8 mmol/L）-treated compared with control group （P〈0.01）. In summary,the high concentration of NEFAs can cause or exacerbate the inflammatory response of calf hepatocytes,may be the factors which cause inflammatory injury to liver of postpartum cows.