摘要
为探索实验室前期改造保存的乳酸菌素PlnJ(G5)对脂多糖LPS诱导下肠上皮细胞IPEC-J2 与小鼠肠道炎症的作用机制.选取肠上皮细胞IPEC-J2 及模式动物小鼠作为研究载体,利用LPS建立炎症模型,并通过添加PlnJ(G5)进行处理,采用cck-8 检测细胞活力以及实时荧光定量PCR检测炎症相关细胞因子的表达变化.结果表明:5.0 μg·mL-1 的LPS对IPEC-J2 细胞活力的影响较小,且该浓度作用下IPEC-J2 细胞IL-1、IL-6、TNF-α、IFN-α及IFN-β的mRNA表达量显著高于空白对照组,证明LPS对IPEC-J2 细胞致炎造模成功;CCK-8 检测结果显示PlnJ(G5)对IPEC-J2 细胞的安全添加剂量为 20.0 μg·mL-1;qPCR检测结果表明IPEC-J2 细胞在经5.0 μg·mL-1 的LPS作用 4h后,添加 20.0 μg·mL-1 未经热处理的PlnJ(G5)对IL-1、IL-6、TNF-α、IFN-α及IFN-β的mRNA表达量的抑制效果最佳;添加乳酸菌素处理对LPS诱导模式动物小鼠肠炎试验发现灌服未经热处理的PlnJ(G5),对小鼠的抗炎效果最好.
Abstract
In order to explore the mechanism of lactobacillus PlnJ(G5)modified and preserved in the laboratory on the intestinal epithelial cells IPEC-J2 and the intestinal inflammation in mice induced by lipopolysaccharide LPS,the intestinal epithelial cell IPEC-J2 and model animal mice were selected as the research carriers.LPS was used to establish an inflammatory model,and PlnJ(G5)was added for the treatment.Then,the cell viability was detected by cck-8 and the expression changes of inflammatory cytokines was detected by the real-time fluorescence quantitative PCR.The results showed that 5.0 μg·mL-1 LPS had little effect on the cell viability of IPEC-J2,and the mRNA expression levels of IL-1,IL-6,TNF-α,IFN-α and IFN-β in IPEC-J2 cells treated with this concentration were significantly higher than those in the blank control group,which proved that the inflammatory model of IPEC-J2 cells induced by LPS was successful.The results of CCK-8 detection showed that the safe additive dosage of PlnJ(G5)for IPEC-J2 cells was 20.0 μg·mL-1.The results of qPCR detection showed that after IPEC-J2 cells treated with 5.0 μg·mL-1 LPS for 4 h,the addition of 20.0 μg·mL-1 unheated PlnJ(G5)had the best inhibitory effect on the mRNA expression of IL-1,IL-6,TNF-α,IFN-α and IFN-β.The experimental results showed that PlnJ(G5),which was not heat treated,had the best anti-inflammatory effect on mice induced by LPS.
维普
万方数据