【摘要】 目的 探讨胆红素对脂多糖诱导的肠上皮细胞保护作用。方法 利用脂多糖（LPS）刺激肠上皮细胞（IEC-6）诱导坏死性小肠结肠炎样（NEC-like）肠上皮损伤，分为对照（Control）组、LPS组、LPS+胆红素（LPS+Bilirubin）组。CCK-8实验检测不同浓度胆红素对LPS诱导IEC-6的影响；ELISA检测炎症因子TNF-α、IL-1β、IL-6和IL-10的表达水平；DCFH-DA检测细胞活性氧（ROS）表达，黄嘌呤氧化酶法测定超氧化物歧化酶（SOD），硫代巴比妥酸法测定丙二醛（MDA）;TUNEL染色检测细胞凋亡；PCNA免疫荧光检测细胞增殖。结果 与Control组比较，LPS组炎症因子TNF-α、IL-1β、IL-6、ROS、MDA表达水平以及TUNEL阳性细胞明显增加（P<0.05）,IL-10、SOD表达水平与PCNA阳性细胞明显降低（P<0.05）；与LPS组比较，LPS+Bilirubin组TNF-α、IL-1β、IL-6、ROS、MDA表达水平以及TUNEL阳性细胞显著下降，IL-10、SOD表达、PCNA阳性细胞显著升高（P<0.05）。结论 胆红素通过降低炎症反应、氧化应激以及促进细胞增殖、抑制细胞凋亡，修复LPS诱导肠上皮细胞损伤，为坏死性小肠结肠炎治疗提供新策略。更多还原

【Abstract】 Objective The aim of this study is to investigate the effect of bilirubin on LPS-induced intestinal epithelial injury.Methods LPS treated intestinal epithelial cell 6 IEC-6 was used as necrotizing enterocolitis-like NEC-like in vitro model. IEC-6were divided into Control group LPS group and LPS + Bilirubin group. CCK-8 reagent was used to detect the effect of different concentrations of bilirubin on injured intestinal epithelium. The expression levels of inflammatory factors TNF-α IL-1β IL-6 and IL-10 were measured using ELISA. Intracellular reactive oxygen species ROS production was assessed by DCFH-DA staining.Superoxide dismutase SOD and malondialdehyde MDA activities were examined by SOD Assay Kit-WST and MDA assay kit-TBA.Apoptosis was evaluated by TUNEL staining and proliferating cell nuclear antigen PCNA were used to assessed proliferative capabilities of intestinal epithelial cells.Results Compared with the Control group the expression levels of TNF-α IL-1β IL-6ROS MDA and TUNEL-positive cells were significantly increased in the LPS group P< 0. 05 and the expression levels of IL-10SOD and PCNA-positive cells were extremely decreased P< 0. 05. In addition the LPS + Bilirubin group showed markedly lower TNF-α IL-1β IL-6 ROS MDA expression levels TUNEL-positive cells and considerably higher IL-10 SOD expression PCNApositive cells than LPS group P< 0. 05.Conclusion Bilirubin was able to repair LPS-induced intestinal epithelial injury by reducing inflammatory response decreasing oxidative stress promoting cell proliferation and inhibiting cell apoptosis which will be a promising therapeutic for NEC.更多还原