肠梗阻对肠上皮Claudin1 mRNA表达的影响

Influence of Intestinal Obstruction on Claudin-1 mRNA Expression of Intestinal Epithelial Cells

目的:探讨急性肠梗阻发生后肠道上皮细胞间紧密连接破坏的时相性特点。方法:选用健康大耳白兔,随机分为正常对照组(Norm)及假手术组(Sham)、梗阻1h组(H1)、梗阻6h组(H6)、梗阻12h组(H12)、梗阻24h组(H24)、梗阻48h组(H48)及梗阻72h组(H72),每组6只。利用实时荧光定量RT-PCR技术,对动物末段小肠上皮的紧密连接相关蛋白Claudin1的mRNA(CLDN1)表达情况进行相对定量分析。结果:梗阻发生后6h发现CLDN1表达的2-△△ct已经显著减少至正常表达量的43%(P<0.05),12h为19%(P<0.01),24h为10%,48～72h阶段表达量很低。结论:肠梗阻发生后早期(6h)已经发生机械屏障的破坏,随着梗阻进展,紧密连接蛋白上游基因的低更加显著,至梗阻48～72h阶段肠上皮的完整性已经完全失去,肠道机械屏障破坏严重。

Objective：To investigate the characteristics of destruction phase of tight junctions in intestinal epithelial cells of acute intestinal obstruction.Methods：Healthy rabbits were randomly divided into normal control group（Norm）,sham-operated group（Sham）,obstruction 1 h group（H1）,obstruction 6 h group（H6）,obstruction 12 h group（H12）,obstruction 24 h group（H24）,obstruction 48 h group（H48） and obstruction 72 h group（H72）,n = 6 for each group.The expression of tight junction-associated protein,Claudin-1 mRNA was analysed by the real-time fluorescence quantitative RT-PCR technology in intestinal epithelial cells.Results：The expression of Claudin-1 mRNA was significantly reduced compared to that of normal（43% for 6 h,P 0.05）,12 h to 19%（P 0.01）,24 h to 10%,which was very low after 48 h-72 h.Conclusion：Mechanical barriers were destroyed at early stage of intestinal obstruction（6 h）.The express of Claudin-1 mRNA was lower with the progress of obstruction.The intestinal epithelial integrity was completely lost;seriously destruction of intestinal mechanical barrier occurred after 48 h-72 h obstruction.