摘要
目的研究血小板活化因子(PAF)对肠上皮细胞屏障通透性的影响,并探讨其作用位点。方法体外培养Caco-2细胞建立肠上皮细胞屏障模型。不同浓度PAF(0,50,100,200 nmol/L)作用24 h,应用TEER和荧光黄透过量检测肠上皮细胞屏障通透性;应用Annexin V和Hoechst染色,用荧光显微镜和流式细胞仪检测肠上皮细胞凋亡情况;透射电镜下观察紧密连接。结果 Caco-2细胞培养21 d左右可形成类似人肠黏膜上皮细胞屏障结构,给予50 nmol/L PAF即可引起TEER的下降和荧光黄透过率的增加,PAF100 nmol/L组肠上皮屏障通透性增加达到高峰,与对照组相比,差异有统计学意义(P<0.01)。实验并未发现PAF引起明显的细胞凋亡增加,但透射电镜下出现了紧密连接的断裂和细胞超微结构的破坏。结论 PAF可直接引起肠上皮细胞屏障通透性增加,该作用是凋亡非依赖性的,作用位点在旁细胞途径。
Objective To study the effect of platelet activating factor（PAF） on intestinal epithelial barrier and the possible target site.Methods The model of intestinal epithelial barrier was established with Caco-2 cells in vitro.The permeability of the intestinal epithelial barrier was detected by transepithelial electrical resistance（TEER） and lucifer yellow flux after the cells were exposed to different concentrations of PAF（0,50,100,and 200 nmol/L） for 24 hours.The cell apoptosis was determined under fluorescence microscope and by flow cytometry after Annexin V and Hoechst staining.The tight junction was observed under transmission electron microscope.Results Caco-2 cells were confluent to form a monolayer after 21 days.TTEER decreased and lucifer yellow flux increased in cells exposed to 50 nmol/L PAF.The permeability of the intestinal epithelial barrier reached the peak in cells exposed to 100 nmol/L PAF（P 0.01）.PAF did not increase cell apoptosis,but tight junction disruption and ultrastructural damage were found under transmission electron microscope.Conclusion PAF could increase the permeability of intestinal epithelial barrier directly,which is independent of apoptosis,and its possible target site is the paracellular pathway.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2011年第3期220-223,共4页
Journal of China Medical University
基金
高等学校博士学科点专项科研基金资助项目(20092104110010)
辽宁省教育厅高校科研计划项目(L2010624)
辽宁省博士启动基金资助项目(2010147)
沈阳市科学技术计划资助项目(F10-205-1-45)
关键词
血小板活化因子
肠上皮细胞屏障
通透性
凋亡
紧密连接
platelet-activating factor
intestinal epithelial barrier
permeability
apoptosis
tight junction
