IL-1β对肠上皮细胞屏障通透性的影响

目的 炎症性肠病是儿童和青少年时期重要的慢性胃肠道疾病,肠黏膜屏障的损伤在其发病中起重要作用.本研究通过IL-β刺激Caco-2细胞单层,体外模拟炎症性肠上皮细胞屏障,为研究炎症性肠病的发病及治疗提供基础.方法 体外培养Caco-2细胞21d模拟肠上皮细胞单层屏障.炎症1组从培养第5天开始每隔1天应用IL-1β处理2h检测TEER值,至第21天.炎症2组于培养第18天换用含IL-1β培养液,分别于处理0、12、24、48、72 h检测TEER.正常对照组用普通培养液培养,也于相应时间点检测TEER.结果 正常对照组细胞TEER从第5天至第15天逐渐增加,在第15天达到600 Ω·cm2,并到达平台期保持至第21 d.炎症1组细胞TEER值均低于正常组细胞,并且直至第21天仍＜500 Ω·cm2.炎症2组细胞显示时间依赖性的TEER逐渐降低,在48 h达到最大值,然后在72 h轻度回升.结论 培养2～3周的Caco-2细胞能够形成具有极性的肠上皮细胞单层,对其给予IL-1β刺激可在体外模拟炎症性肠上皮细胞屏障.

The effects of IL-1 beta on intestinal epithelial barrier permeability

Objective Inflammatory bowel disease is an important chronic gastrointestinal disease of childhood and adolescence.Intestinal mucosa barrier damage plays an important role in its pathogenesis.This study attempts to use IL-1β stimulating Caco-2 cell monolayer simulates inflammatory intestinal epithelial barrier in vitro,provides the basis for studying the pathogenesis and treatment of IBD.Methods Caco-2 cells were cultivated in vitro until the 21st day to simulate intestinal epithelial cell monolayer barrier.The cells of inflammation group one were disposed with IL-1β for 2 h since the 5th day and detected TEER every other day until the 21st day.The cells of inflammation group two were disposed with IL-1 β at the 18th day for 0,12,24,48,72h, and detected TEER respectively.Normal control group cells were cultured with common medium and detected TEER at the corresponding time point.Results The TEER of Caco-2 cells gradually increased from the 5th to 15th days,reached 600Ω·cm2 in the 15th day of a plateau until to the 21st day.Since the 5th day,the TEER of inflammation group one were all lower than normal group,and still to the 21st day ＜ 500Ω·cm2.Inflammation group two shows the time dependence TEER gradually reduce,peaked at 48 hours,then a slight increase in 72 hours.Conclusion The Caco-2 cells cultured for 2 ～ 3 weeks can form intestinal epithelial monolayer barrier with polarity,then treated with IL-1 β can manufacture inflammatory intestinal epithelial barrier model in vitro.