植物乳杆菌对IL-10基因敲除结肠炎小鼠黏附分子的影响  

Lactobacillus plantarum attenuates up-regulation of adhesion molecules expression in IL-10 gene knockout mice with colitis

作　　者：储昭新[1] 陈红旗[1] 张明[1] 秦环龙[1] 

机构地区：[1]上海第六人民医院普外科,上海市200233

出　　处：《世界华人消化杂志》2009年第12期1190-1196,共7页World Chinese Journal of Digestology

基　　金：国家重点基础研究计划(973)基金资助项目;No.2008CB517403;黎介寿院士肠道屏障研究专项基金资助项目;No.LJS_2009001~~

摘　　要：目的:观察植物乳杆菌(Lactobacillus plantarum,LP)灌胃对IL-10基因敲除小鼠肠道炎症和淋巴细胞归巢的影响.方法:取IL-10基因敲除(knockout,KO)小鼠和未作基因敲除的背景鼠分为4组:对照组(野生组WT)、加植物乳杆菌组(WT+LP)、IL-10基因敲除模型组(KO)、模型加植物乳杆菌组(KO+LP).4wk开始对照组和KO组每日予PBS灌胃,WT+LP和KO+LP组予溶于PBS的LP灌胃,持续4-8wk结束.实验结束后取各组小鼠结肠行炎症评分和电镜亚显微结构观察,并用RT-PCR和Western blot检测归巢相关分子MAdCAM-1、ICAM-1、α4β7及CD3的表达.结果:8wk后KO小鼠100%发生肠道炎症,且其CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较WT组均明显增高(mRNA:t=39.42,8.83,25.53,45.78,均P<0.01;CD3、ICAM-1、MAdCAM-1蛋白:t=19.04,29.57,12.29,均P<0.01).予以益生菌LP灌胃后,KO+LP组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA和蛋白表达水平较KO组均明显降低(mRNA:t=20.34;4.95;14.21;22.31,均P<0.01;CD3、ICAM-1、MAdCAM-1蛋白:t=6.82,14.10,7.03,均P<0.01);WT+LP组小鼠CD3及黏附分子α4β7、ICAM-1和MAdCAM-1的mRNA较WT组均明显降低(t=9.33,10.55,7.75,6.69,均P<0.01),而WT+LP组小鼠CD3及黏附分子ICAM-1和MAdCAM-1的蛋白表达水平无明显降低.结论:植物乳杆菌能下调黏附分子在IL-10基因敲除结肠炎小鼠中的高表达,这可能是其减轻炎症状态,缓解炎症性肠病的重要机制之一.AIM： To investigate the effect of lactobacillus plantarum （LP） daily intragastric administration on lymphocyte homing and intestinal inflammation in IL-10 gene knockout mice colitis. METHODS： Both two groups of 10 IL-10 KO mice and two groups of normal background controls were consecutively fed on phosphate buffered saline containing LP 1258 at 10^12 CFU/L, and unmodified phosphate buffered saline, respectively, from 4 to 8 wk. At sacrifice, the bowels were histologically scored under light microscope and observed under transmission electron microscope. Expression of CD3 and intestinal homing-related molecules MAd- CAM-1, ICAM-1,α4β7 were detected by reversetranscription polymerase chain reaction and Western blot. RESULTS： At 8 wk, 100% of the IL-10 gene-de- ficient mice showed histological inflammation. CD3, adhesion molecules α4β7, ICAM-1 and MAdCAM-1 had a significantly higher expression of both mRNA and protein levels in KO than WT group （mRNA： t = 39.42, 8.83, 25.53, 45.78, all P 〈 0.01; CD3, ICAM-1, MAdCAM-1 protein： t = 19.04, 29.57, 12.29, all P 〈 0.01）. After LP treatment, the expression of both mRNA and protein was significantly reduced in KO＋LP, compared with KO mice （mRNA： t = 20.34, 4.95, 14.21, 22.31, all P 〈 0.01; CD3, ICAM-1, MAd- CAM-1 protein： t = 6.82, 14.10, 7.03, all P 〈 0.01）. The expression of mRNA was significantly reduced in WT＋LP, compared with WT mice （t = 9.33, 10.55, 7.75, 6.69, all P 〈 0.01）, however, protein expression was not significantly reduced. CONCLUSION： LP can interfere with upregulation of adhesion molecules, attenuating colitis of IL-10 gene knockout mice.

关 键 词：益生菌 植物乳杆菌 炎症性肠病 白介素-10 基因敲除 淋巴细胞归巢 

分 类 号：R574.62[医药卫生—消化系统]