小檗碱改善肠黏膜屏障损伤及其可能机制的实验研究  

Experimental Study of Berberine Improving Intestinal Mucosal Barrier Damage and Its Possible Mechanisms

作　　者：邹小慧 陆付耳[1] 舒婷婷 刘浩[2] 

机构地区：[1]华中科技大学同济医学院,湖北武汉430030 [2]华中科技大学附属梨园医院,湖北武汉480077

出　　处：《武汉大学学报（医学版）》2018年第2期207-212,218,共7页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:NSFC2014)

摘　　要：目的：研究小檗碱（BBR）改善肠黏膜屏障损伤及可能机制。方法：人结肠腺癌细胞系（caco-2）培养形成完整的紧密连接单层。不同浓度的脂多糖（LPS）（0,0.1,1,10μg/ml）作用于caco-2细胞24h后,检测单层跨膜电阻与通透性以及凋亡相关蛋白caspase12表达。实验分为对照组、模型组（LPS 10μg/ml）,干预组（LPS 10μg/ml与BBR 10μmol/L）,作用24h后检测通透性与紧密连接蛋白zo-1,occludin,claudin-1的表达以及各组TLR4,MyD88,NF-κB的变化。结果：caco-2细胞单层通透性在LPS（0-10μg/ml）范围内呈浓度依赖性增加;10μg/ml以下浓度的LPS作用后caspase12表达与对照组无明显差异;与对照组比较,模型组的紧密连接蛋白表达明显降低,蛋白TLR4,MyD88,NF-κB的表达明显增加（P〈0.05）;而与模型组比较,干预组紧密连接蛋白则明显增加,蛋白TLR4,MyD88,NF-κB的表达明显降低（P〈0.05）。结论：小檗碱能增加肠上皮细胞间紧密连接（TJ）蛋白的表达,从而有效修复LPS引起的肠黏膜屏障损伤,LPS诱导caco-2细胞紧密连接蛋白减少可能是通过TLR4,MyD88,NF-κB信号通路的活化,而BBR能抑制该信号通路从而能改善紧密连接蛋白的表达。Objective： To study the protective effect of berberine on the intestinal mucosal barrier func- tion damage and its possible mechanisms. Methods： Caco-2 cell were cultured into integrated tight junction monolayer, then stimulated with different concetration of LPS （0,0.1,1,10μg/ml） for 24 hours. Transepithelial electrical resistance, paracellular permeability and the expression level of caspase12 were measured. Experiments were divided three groups： control group, model group （LPS10μg/ml） and treatment group （LPS 10 μg/ml＋BBR 10 μmol/1）. The change of monolay- er permeability and the expression level of tight junction（TJ） proteins including zo-1, occludin and claudinl were observed, then, TLR4, MyD88 and NF-κB were also measured. Results. Caco-2 cell monolayer permeability increased with the increase of the concentration of LPS （0-10μg/ml）, but the expression level of caspasel2 in model group has no difference compared with that in control group. Compared with control group, the expression level of TJ proteins in model group de- creased obviously, while the expression level of TLR4,MyD88 and NF-κB significantly increased （P〈0. 05）. Furthemore, compared with model group, the expression level of TJ proteins in treatment group increased remarkably, while the expression level of TLR4,MyD88 and NF-κB in treatment group had sharp decreases （P〈0.05）. Conclusion： Berberine can effectively improve the LPS-induced intestinal mueosal barrier injury by increasing the expression of TJ proteins. TJ proteins decrease induced by LPS may via activating TLR4/MyD88/NF-κB signal pathway, while berberine inhibits this signal pathway.

关 键 词：小檗碱 CACO-2细胞 脂多糖 肠黏膜屏障 紧密连接蛋白 

分 类 号：R587.1[医药卫生—内分泌]