IL-22改善脂多糖诱导的Caco-2单层肠上皮紧密连接损伤  

IL-22 ameliorates lipopolysaccharide-induced intestinal epithelial tight junction injury in Caco-2 monolayers

作　　者：李健[1] 唐富琴 孔大陆 胡均 LI Jian;TANG Fu-qin;KONG Da-lu;HU Jun(Tianjin University Hospital,Tianjin 300072,China;Division of Cardiovascular Surgery,Xinqiao Hospital,Army Medical University,Chongqing 400037,China;Department of Colorectal Cancer Surgery,National Clinical Research Center for Cancer,Key Laboratory of Cancer Prevention and Therapy of Tianjin,Tianjin’s Clinical Research Center for Cancer,Tianjin Medical University Cancer Institute and Hospital,Tianjin 300060,China)

机构地区：[1]天津大学医院,天津300072 [2]陆军军医大学新桥医院心血管外科,重庆400037 [3]天津医科大学肿瘤医院结直肠肿瘤科,国家肿瘤临床医学研究中心,天津市肿瘤防治重点实验室,天津市恶性肿瘤临床医学研究中心,天津300060

出　　处：《生物医学工程与临床》2020年第4期456-461,共6页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(811018700);国家临床重点专科建设项目(2013-544);天津市卫健委重点项目(16kg127);CSCO-恒瑞肿瘤研究基金资助项目(Y-HR2017-073)。

摘　　要：目的紧密连接(TJ)屏障功能障碍在炎症性肠病(IBD)中起重要作用。据报道IL-22对IBD有直接的保护作用。研究IL-22对TJ屏障功能的保护作用。方法选择肠上皮Caco-2单层细胞,分为对照组、脂多糖(LPS)组、IL-22组、LPS+IL-22组。采用LPS(100μg/mL)或IL-22(100 ng/mL)处理肠上皮Caco-2单层细胞0、12、24、48 h。Western blot检测IL-22受体IL-22R1和TJ蛋白(ZO-1和Occludin)表达。检测Caco-2单层细胞膜跨膜电阻(TEER)。分析Caco-2单层细胞被LPS、IL-22处理48 h后的细胞膜TEER、TJ蛋白变化和免疫荧光方法观察TJ蛋白在细胞膜的分布。结果LPS可上调Caco-2细胞中IL-22R1的表达。LPS可显著降低Caco-2单层细胞膜TEER(刺激24 h,下降水平接近50%),差异有显著统计学意义(P<0.001)。IL-22组ZO-1、Occludin表达水平高于LPS组、LPS+IL-22组,差异有显著统计学意义(P<0.01)。LPS+IL-22组ZO-1、Occludin表达水平高于LPS组,差异有显著统计学意义(P<0.01)。免疫荧光法检测可见,对照组和IL-22组ZO-1、Occludin位于细胞膜,呈明亮连续条带;LPS组,位于细胞膜的ZO-1、Occludin明亮带变暗,显示强度和间断度降低;LPS+IL-22组细胞膜可见ZO-1、Occludin明亮带,但明亮带略弱于对照组和IL-22组。结论IL-22在LPS诱导的肠屏障功能紊乱中起保护作用,并为IL-22在IBD中如何促进肠上皮屏障的完整性提供了新的证据。Objective To study the protective effect of interleukin(IL)-22 on tight junction(TJ)barrier function,based on the fact that TJ barrier dysfunction plays an important role in inflammatory bowel disease(IBD)and IL-22 shows direct protective effect on IBD.Methods Intestinal epithelial Caco-2 monolayer cells were selected and divided into control group,lipopolysaccharide(LPS)group,IL-22 group and LPS+IL-22 group.LPS(100μg/mL)or IL-22(100 ng/mL)was used to treat Caco-2 monolayers for 0,12,24 and 48 hours,respectively.Western blot was used to detect IL-22 receptor IL-22 R1 and TJ protein(ZO-1 and Occludin)expression.The Caco-2 monolayer cell membrane trans-epithellal electric resistance(TEER)was detected.The changes of TEER and TJ proteins in Caco-2 monolayer cell treated with LPS and IL-22 for 48-hour were analyzed,and immunofluorescence was used to observe distribution of TJ protein in cell membrane.Results LPS up-regulated IL-22 R1 expression in Caco-2 cell.LPS significantly reduced TEER of Caco-2 monolayers(stimulated for 24-hour and decreased close to 50%),and the difference was statistically significant(P<0.001).The expression levels of ZO-1 and Occludin in IL-22 group were higher than those in LPS group and LPS+IL-22 group,and the differences were statistically significant(P<0.01).The expression levels of ZO-1 and Occludin in LPS+IL-22 group were higher than those in LPS group,and the differences were statistically significant(P<0.01).The immunofluorescence detection results showed that ZO-1 and Occludin in control group and IL-22 group were located on cell membrane with bright continuous bands.ZO-1 and Occludin bright bands in cell membrane of LPS group was darkened,the intensity and discontinuity were reduced;ZO-1 and Occludin bright bands were observed in LPS+IL-22 group on cell membrane,but the bright bands were slightly darker than that in control group and IL-22 group.Conclusion It is demonstrated that IL-22 plays protective role in disruption of intestinal barrier function induced by LPS,which provides n

关 键 词：IL-22 肠上皮屏障 紧密连接 屏障功能 炎症性肠病 

分 类 号：R574[医药卫生—消化系统] R364.5[医药卫生—内科学]