摘要
目的观察缺氧/复氧(hypoxia/reoxygenation,H/R)损伤条件下大黄素(Emodin)对肠上皮细胞表达紧密连接蛋白Occludin和ZO-1的影响,以研究大黄素对肠黏膜屏障的保护作用机制。方法体外培养肠上皮细胞株Caco-2建立H/R模型,将细胞随机分为正常细胞(N)组、缺氧复氧(H/R)组、缺氧复氧+Hank平衡盐溶液(H/R+HBSS)组、缺氧复氧+大黄素(H/R+EN)组。检测跨上皮细胞电阻(transepithelial electrical resistance,TEER)值;使用透射电子显微镜观察紧密连接的变化;用Western blot法测定Occludin和ZO-1蛋白的表达水平;半定量RT-PCR法测定Occludin和ZO-1的mRNA表达水平。结果与H/R组、H/R+HBSS组相比,H/R+EN组的紧密连接破坏明显减少;H/R+EN组的TEER值、Occludin和ZO-1的蛋白及mRNA表达水平明显高于H/R组、H/R+HBSS组(P<0.05)。结论大黄素可通过减少Occludin、ZO-1的破坏并增加其表达来保护肠黏膜屏障。
Objective Through observing the effects of emodin on the tight junction（TJ）protein Occludin and ZO-1 of intestinal epithelial cells damaged by hypoxia/reoxygenation（H/R）,to study the mechanism of the protective effects of emodin on the intestinal mucosa barrier.Methods Bowel epithelial cell lines Caco-2 cells were cultured in vitro to establish H/R model.The cells were averagely randomized into four groups ：N（normal cells）group,H/R（hypoxia/reoxygenation）group,H/R＋HBSS（hypoxia/reoxygenation and Hanks balanced solution）group,H/R＋EN（hypoxia/ reoxygenation and Emodin）group.The transepithelial electrical resistance（TEER）value was measured;transmission electron microscope was used to observe the changes of TJ;Western blot analysis was employed to detect the expression level of Occludin and ZO-1 protein;Semi-quantitative reverse transcription polymerase chain reaction（RT-PCR）was used to detect the mRNA expression level of Occludin and ZO-1.Results Comparing with the H/R group and H/R＋HBSS group,the damage extent of TJ was significantly reduced in the H/R＋EN group.The TEER value,the protein and mRNA expression level of Occludin and ZO-1 in the H/R＋EN group were obviously higher than that of the H/R group and H/R＋HBSS group（P〈0.05）.Conclusion Emodin can decrease the damage of Occludin and ZO-1,and increasing their expression to protect the intestinal mucosa barrier.
出处
《东南国防医药》
2012年第3期199-202,共4页
Military Medical Journal of Southeast China
基金
南京军区医药卫生科研基金课题(09Z027)
