丁酸钠在Caco-2细胞肠屏障中的作用及p38MAPK的影响  

Effects of sodium butyrate on the intestinal barrier function in a Caco-2 cell monolayer model of intestinal barrier

作　　者：黄晓忠[1] 林正秀[1] 张田丰[1] 朱利斌[1] 李仲荣[1] 林锦[2] 

机构地区：[1]温州医学院附属第二医院、育英儿童医院小儿外科,325027 [2]纽约大学西奈山医学院

出　　处：《中华小儿外科杂志》2013年第6期458-462,共5页Chinese Journal of Pediatric Surgery

基　　金：浙江省自然科学基金（编号：LY12H04005）;浙江省医药卫生支撑学科基金（编号：11-ZC27）

摘　　要：目的观察不同浓度丁酸钠对Caco-2细胞肠屏障模型的作用情况及p38MAPK在这一过程中可能的影响，为进一步纠正肠屏障功能异常寻找新靶点。方法通过Caco-2细胞培养，建立单层Caco-2细胞肠屏障模型；比较对照组、不同浓度丁酸钠（2mmol／L、5mmol／L和8retool／L）及丁酸钠（5mmol／L）＋SB203580（p38MAPK特异性抑制剂，20μmol／L）组对Caco-2细胞肠屏障模型的作用情况，测定连续培养72h后肠屏障模型的跨上皮电阻（TER）、菊粉一FITC（inulin-FITC）的通透性及扫描电镜表现。结果体外培养的Caco-2细胞能在3周后形成肠上皮细胞屏障模型。低浓度丁酸钠（2mmol／L）作用于肠屏障模型72h后，TER达到（467．65±7．41）Ω·cn-i2，与对照组的（417．58±6．29）Ω·cn-i2相比差异有统计学意义（Pd0．05）；中、高浓度丁酸钠（5mmol／L和8mmol／L）作用72h后，TER分别为（113．87±20．27）Ω·cm2和（58．60±29．02）Ω·cm2，菊粉通透性分别为（11．08±1．04）％和（16．01±1．56）％，与对照组相比差异均有统计学意义（P〈0．05）。而5mmol／L丁酸钠＋20ffmol／LSB203580组TER和通透性分别为（395．17±9．97）Ω·cm2和（7．19±0．97）％，与单纯5mmol／L丁酸钠组相比，差异有统计学意义（P〈O．05）。扫描电镜发现对照组和2mmol／L丁酸钠组单层Caco-2细胞屏障表面完整，细胞连接紧密无脱落；5mmol／L和8mmol／L丁酸钠组随药物浓度增加，细胞变形呈球形脱落亦增加；5mmol／L丁酸钠＋20／,mol／LSB203580组细胞脱落较5mmol／L丁酸钠组明显减轻。结论丁酸钠对Caco-2细胞肠屏障模型作用具有双向性：低浓度丁酸钠促进肠屏障功能，而中、高浓度丁酸钠可破坏肠屏障功能；p38MAPK特异性抑制剂SB203580可减轻中等浓度丁酸钠对肠屏障功能的损害。Objective To explore the effects of different concentrations of sodium butyrate on a Caco-2 cell monolayer model of intestinal barrier and to investigate the potential influence of p38 MAPK on this process. Methods The Caco-2 cell monolayer model of intestinal barrier was estab- lished via cell culture,which was further divided into 3 groups including the control group, the sodium butyrate groups with different concentrations （2 mmol/L,5 mmol/L,and 8 mmol/L）） and the sodium butyrate （5 mmol/L）± SB203580（20/amol/L , p38MAPK specific inhibitor）group. After 72 hours of culture,the transepithelial electrical resistance （TER） and the barrier permeability by testing the con- centration o{ inulin-FITC were detected. The surface of Caco-2 cells was observed by electronic scan- ning microscopy. Results After 3 weeks of culture, monolayer Caco-2 ceils formed the intestinal epi- thelial cell barrier model. After incubation of sodium butyrate （2 mmol/L） on Caco-2 cell monolayers for 72 h,the TER was increased to （467.65 ± 7. 41） Ω·cm2 compared to （417. 58 ± 6. 29）Ω·cm2 in the control group （P〈0. 05）. After incubation of sodium butyrate （5 mmol/L or 8 mmol/L） on Caeo-2 ceils monolayer for 72 h,the TERs were （113.87 ± 20. 27） Ω·cm2 and （58. 60 ± 29. 02）Ω·cm2 ,which were both significantly lower than those in the control group （P〈0. 05）. The inulin-FITC permeabilitywas （ 11.08 ± 1.04） and （16. 01 ± 1. 56） in these 2 groups （P〈0. 05, compared to the control group） . However, the TER and permeability of the 5 mmol/L sodium butyrate ± 20 μmol/L SB203580 group were （395. 17 ± 9. 97）Ω·cm2 and （7. 19 ± 0. 97）% （both P〈0. 05, compared to those in the 5 nunol/L sodium butyrate group）. The electronic microscopy revealed that the surface of the Cocci2 cells,which were treated with 0 mmol/L and 2 mmol/I, sodium butyrate, was connected tightly with out cell float. The surface impairment and cell float of Caco-2 cell monolayers were incre

关 键 词：脂肪酸类 挥发性 肠黏膜 丝裂原激活蛋白激酶类 

分 类 号：R722[医药卫生—儿科]