Toll样受体2对肠上皮细胞屏障通透性的保护作用及机制  

The protective effect and mechanism of TLR2 on permeability of intestinal epithelial barrier

作　　者：林楠[1] 许玲芬[1] 滕旭[1] 孙梅[1] 

机构地区：[1]中国医科大学附属盛京医院儿科,沈阳110004

出　　处：《国际儿科学杂志》2016年第6期484-487,501,共5页International Journal of Pediatrics

基　　金：国家自然基金青年科学基金（81400585）;辽宁省自然科学基金（2014021042）;沈阳市科技计划项目（F13-316-1-71）

摘　　要：目的肠上皮细胞屏障的损伤与多种胃肠道疾病的发生密切相关，有效维持屏障功能是治疗这些疾病的关键。本研究试图通过体外实验探讨Toll样受体2（toll—like receptor，TLR2）对肠上皮细胞屏障通透性的保护作用及其机制。方法正常组将未转染Caco-2细胞，TLR2基因沉默Caco-2细胞，TLR2基因过表达Caco-2细胞培养21d形成单层，检测跨膜电阻（transepithelial electrical resistance,TEER）值，其反应上皮细胞屏障的通透性。炎症组3类细胞分别于培养第19d给予10ng／ml IL-1β刺激48h，于第21d检测TEER值。抑制剂组3类细胞分别在1L-1β前再给予PI3K／Akt通路抑制剂处理1h，于第21d检测TEER值。结果TLR2基因沉默能够显著减低Caco-2细胞单层的TEER值（P〈0．01），而TLR2基因过表达能够升高TEER值，但不具有统计学意义。TLR2能够防止IL-1β造成的TEER值下降（P〈0．01），此作用在给予PI3K／Akt通路抑制剂后消失。结论TLR2对肠上皮细胞屏障通透性具有调节作用，并且能够防止炎症造成的通透性增高，这种保护作用由PI3K／Akt通路参与介导。Objective Intestinal epithelial barrier damage is closely related to a variety of gastrointesti- nal disease, how to maintain is function effectively is the key to treat all these diseases. This research attempts to explore the protective effect and its mechanism of toll-like receptor 2 （ Toll -like receptor, TLR2） on permeability of intestinal epithelial barrier by experiments in vitro, to lay a foundation for new treatment methods. Methods We cultured non-transfected Caco-2 cells, TLR2-deficiency Caco-2 cells, TLR2-overexpressed Caco-2 ceils in normal control group until the 21 st d, then tested transepithelial electrical resistance （TEER）which reacts the permeability of epithelial barrier. We cultured 3 types of ceils in inflammation group until the 19th d treated with 10 ng/ml IL-1 beta for 48 h,then tested TEER values at the 21st d. We treated 3 types of cells in inhibition group with PI3K/Akt pathway inhibitor for lh befor IL-1 beta,then tested TEER values at the 21st d. Results TEER value of TLR2-deficiency Caco-2 cell monolayer significantly reduced （ P 〈 0. 01 ）, whereas TEER value of TLR2-overexpressed Caco-2 monolayer raised, but without statistically significant. TLR2 can prevent IL-1 beta caused TEER decreasing（ P 〈 0. 01 ） ,but the effect disappeared after given PI3K/Akt pathway inhibitor. Condusion TLR2 can regulate the permeability of intestinal epithelial barrier. In addition, TLR2 can protect permeability increasing caused by inflammation,this effect mediated by PI3K/Akt pathway.

关 键 词：肠上皮细胞屏障 跨膜电阻 TLR2 PI3K/AKT通路 炎症性肠病 

分 类 号：R574[医药卫生—消化系统]