益生菌VSL#3对实验性结肠炎大鼠结肠黏膜Claudins蛋白表达的影响  

The effects of probiotics VSL# 3 on the expression of claudins in the colon mucosa of experimental colitis rats

作　　者：张峻[1] 罗燕[1] 朱尤庆[1] 

机构地区：[1]武汉大学中南医院消化科 湖北省肠病医学临床研究中心,430071

出　　处：《中华消化杂志》2011年第9期613-618,共6页Chinese Journal of Digestion

摘　　要：目的观察益生菌VSL#3对2，4，6-三硝基苯磺酸（TNBS）诱导的大鼠急性结肠炎的保护作用，以及对紧密连接蛋白Claudins（Claudin-1，-2，-3）在结肠黏膜上表达的影响。方法24只健康雌性SD大鼠采用TNBS一次性灌肠法建立大鼠结肠炎模型，造模完成后第2天开始，8只大鼠每日给予2ml 0．9％NaCI溶液灌胃（TNBS组），8只给予100mgVSL#3（VSL#3组），另8只给予200mg5氨基水杨酸（5-ASA）灌胃（5-ASA组）。5只同级别未造模大鼠作为对照组每日给予0．9％NaCl溶液灌胃，均持续1周并处死。处死时留取粪便标本并做细菌培养以检测肠道菌群的变化。通过观察腹泻、便血和体重情况计算疾病活动指数（DAI）。取结肠组织做HE染色，观察病理学改变；酶联免疫吸附试验（ELISA）检测肠组织髓过氧化物酶（MPO）的变化；量子点免疫荧光标记法检测Claudins在结肠黏膜的表达和分布。结果与TNBS组比较，益生菌VSL#3和5-ASA治疗均能降低结肠炎大鼠明显升高的DAI和肠组织MPO水平，减轻结肠炎性反应评分。与对照组比较，结肠炎大鼠的肠道细菌计数发生改变，Claudin-1，-2，-3蛋白表达均明显增加（平均荧光强度分别为66．200±5．737、71．780±6．670、61．300±5．199，t值分别=17．237、27．909和21．788；P值均〈0．01）；而与TNBS组和5ASA组比较，VSL#3可调节肠道菌群平衡，增加Claudin-1，3的表达（Claudin-1：75．550±8．717比66．200±5．737和67．080±5．401；t=9．348，8．469；P值均〈0．05；Claudin-3：68．820±7．443比61．300±5．199和59．830±5．930；t=7．519，8．988，P值均〈0．05），降低Claudin-2的表达（58．740±6．457比71．780±6．670和66．870±5．791；t=13．033，P〈0．01；t=8．123，P〈0．05）。结论益生菌VSL#3可能通过改变Claudins蛋白的表达，增强肠黏膜屏障功能，从而发挥缓解急性结肠炎性反应的作用。Objective To observe the protective effect of probiotics VSL # 3 on the rats of trinitrobenzene sulphonic acid （TNBS） induced acute colitis, and the effect on the expression of tight junction proteins elaudins （claudinl, 2, and 3） in colon mucosa. Methods Rat of colitis model was induced by TNBS one time enema. Since the second day after the model established, 2 ml 0. 9% sodium chloride was intragastric administrated into 8 rats each day （TNBS group）, 8 rats with 100 mg VSL# 3 （VSL# 3 group）, another 8 rats with 5-aminosalicylic acid （5-ASA） intragastric administration （5 ASA group）, and 0. 9% sodium chloride was Jntragastric administrated daily into 5 no modeling rats as control group. All were treated for 7 days and then sacrificed. Fecal samples were collected for bacterial culturing and detecting the changes of colon bacteria colonies. Disease activity index （DAI） was calculated according to diarrhea, hemafecia and weight. The colon tissue was taken for HE staining and the pathological changes were observed under microscope. The changes of myeloperoxidase （MPO） level in colon tissue were examined by enzyme-linked immunosorbent assay （ELISA）. The expression and distribution of claudins 1, 2, and 3 in colon tissue was detected by a novel quantum dots immunofluorescence labeling technique. Results Compared with TNBS group, both VSLg 3 and 5-ASA treatment could improve colitis, as well as deereas DAI and colonic MPO level, and lower the inflammatory score of colitis. Compared with control group, the bacteria counting in the colon of colitis rats changed, the expression of Claudins 1, 2, and 3 expression was significantly increased （average density.. 66. 200±5. 737, 71. 780±6. 670 and 61. 300±5. 199,t=17. 237, 27. 909 and 21. 788, all P〈0.01）. Compared with TNBS group or 5-ASA group, VSL# 3 could adjust the balance of intestinal bacteria and increase the expression of Claudin 1 and 3 （Claudin 1： 75. 550± 8. 717, 66. 200± 5. 737 and 67. 080±5.401;t=9. 348, 8. 469;

关 键 词：连接蛋白类 肠黏膜 益生菌 三硝基苯磺酸 炎症性肠病 量子点 荧光抗体技术 

分 类 号：R574.62[医药卫生—消化系统]