摘要
目的:了解肿瘤坏死因子(TNF-α)和干扰素(IFN-γ)对上皮细胞功能的作用,研究TNF-α和IFN-γ与上皮细胞屏障功能的关系。方法:T84细胞株用加有5%胎牛血清的DMEM/F-12混合培养液培养。实验分为四组:即对照组、TNF-α处理组、IFN-γ处理组和TNF-α与IFN-γ联合处理组。TNF-α处理组和IFN-γ处理组细胞分别用加有10ng/mLTNF-α和100U/mLIFN-γ的培养液处理72h。各细胞因子联合处理组用100U/mLIFN-γ处理19h后,再加入10ng/mLTNF-α。用EVOM电压电阻仪测定细胞跨膜电阻抗(TER)变化,用透射电镜观察细胞紧密连接的形态改变。结果:IFN-γ单独处理48h后,TER降低至78.06%,培养至72h降低为59.31%。在单一TNF-α作用下,24h后TER值逐渐降低,72h降低至56.82%。与对照细胞相比,TNF-α和IFN-γ联合处理的T84细胞株TER值显著降低,72hTER值为20.88%,表明细胞紧密连接的结构破坏。结论:TNF-α和IFN-γ损伤T84细胞株的紧密连接屏障功能,两者具有协同作用。
Objective：Inflammatory bowel disease （IBD） is characterized by chronic intestinal inflammation and increased epithelial permeability. Both tumour necrosis factor-α （TNF-α） and interferon-γ （IFN-γ） have been implicated in IBD. To understand better disruption of epithelial barrier function by these cytokines, we investigated the effects of IFN-γand TNF-α on intestinal epithelium tight junction. Methods ：T84 cells were grown in a 1 ： 1 mixture of DMEM and F-12 medium supplemented with 5% heat-inactivated fetal bovine serum. The cell monolayers were incubated with IFN-γ （ 100 U/mL）, TNF-α （ 10 ng/mL） or the combination of IFN-γpreincubated（ 100 U/mL） and TNF-α （ 10 ng/mL）. Control cells were cultured in the absence of cytokines. Transepithelial resistance （TER） was measured using an EVOM voltohmmeter. Changes in the morphology of TJ structures were observed by transmission electron microscopy, r Results：We found that TER was slightly changed in 72 hours of culture with basolateral IFN-γor TNF-α. After incubation with IFN-γfor 48 hours TER reduced to 78.06%, and TER decreased to 59.31% at 72 h. TER decreased gradually after 24 hours incubation with TNF-α and it reduced to 56.82% at 72 h. Compared with controls, significant changes occurred after culture with IFN-γ plus TNF-α, TER was decreased to 20.88% and the structure of TJs was disrupted. Conclusion：TNF-α and IFN-γsynergistically affected the epithelial barrier and disrupted the structure of TJs.
出处
《肠外与肠内营养》
CAS
2006年第6期335-337,共3页
Parenteral & Enteral Nutrition
基金
国家973项目(2003CB515502)
军队科研基金面上项目(0603AM117)
南京军区医学科学技术研究"十一五"计划(06Z40)
关键词
紧密连接
细胞因子
跨膜电阻抗
超微结构
Tight junction
Cytokines
Transepithelial resistance
Ultrastructure
