TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响  

TLR2McAb and TLR4McAb decrease inflammatory cytokines expression and increase counts of Lactobacilli and Bifidobacteria in dextran sulfate sodium-induced colitis in mice

作　　者：董乐[1] 刘懿[1] 李鹃[1] 岳文杰[1] 

机构地区：[1]复旦大学附属华山医院消化科,上海200040

出　　处：《中华微生物学和免疫学杂志》2010年第6期532-538,共7页Chinese Journal of Microbiology and Immunology

基　　金：上海市科委基础研究重点课题（09JC1402900）

摘　　要：目的 研究Toll样受体2单克隆抗体（Toll-like receptor 2 monoclonal antibody,TLR2McAb）、Toll样受体4单克隆抗体（TLR4McAb）对葡聚糖硫酸钠（dextran sulfate sodium,DSS）诱导的小鼠急性期溃疡性结肠炎组织学改变及肠道菌群结构的影响.方法 雄性BALB/c小鼠分为正常对照组（A）、急性期溃疡性结肠炎模型组（B）、TLR2McAb干预组（C）、TLR4McAb干预组（D）、TLR2McAb＋TLR4McAb共同干预组（E）,每组10只.A组饮用蒸馏水,B～E组饮用5%DSS水溶液以产生急性期溃疡性结肠炎,造模开始同时,每隔48 h予A、B两组小鼠腹腔内注射生理盐水,C～E组小鼠分别予以10μg的TLR2McAb、TLR4McAb、TLR2McAb＋TLR4McAb,期间观察小鼠的疾病活动指数（disease activity index,DAI）.干预7 d后处死小鼠,HE染色观察结肠炎症,并进行结肠组织病理学评分（histopathological score,HS）;使用real-time PCR法检测各组结肠黏膜中IFN-γ、IL-4、IL-17表达;留取盲肠内粪便对大肠杆菌、双歧杆菌及乳酸杆菌进行培养并比较各菌群菌落计数（colony forming units,CFU）的变化.应用SPSS13.0软件进行统计学分析,以P〈0.05为差异有统计学意义.结果 （1）与A组比较,B组的DAI及HS显著增高（DAI：9.700±2.406 vs 0.500±0.707,P〈0.01;HS：16.500±2.991 vs 6.400±3.273,P〈0.01）;C、D组与B组比较DAI及HS差异均无统计学意义（P〉0.05）;E组的DAI及HS明显低于B组（DAI：5.700±3.498 vs 9.700±2.406,P〈0.05;HS：9.500±3.308 vs 16.500±2.991,P〈0.01）.（2）与A组比较,B组小鼠结肠黏膜IFN-γ、IL-4及IL-17的mRNA表达明显增高;而C～E组结肠黏膜三种细胞因子的表达均有不同程度的降低.（3）A组大肠杆菌数量较少,B～E四组大肠杆菌均明显生长（与A组比较P〈0.05）,C～E组大肠杆菌数值与B组比较有所下降,但差异无统计学意义（P〉0.05）;B组双歧杆菌及乳酸杆菌数量均明显少于A组及C～E组（P〈0.05）,使用TLR2McAb、TLR4McAb�Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice （SPF level）, were randomly assigned into five groups： the control group（ group A）, the UC model group（ group B）, TLR2McAb intervention group（ group C）, TLR4McAb intervention group（ group D） and TLR2McAb ＋ TLR4McAb intervention group（group E）. Clinical symptoms were evaluated by the disease activity index（DAI）, while tissue sam ples were evaluated by histological scoring（HS）. The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.

关 键 词：溃疡性结肠炎 TLR2单克隆抗体 TLR4单克隆抗体 肠道菌群 

分 类 号：R574.62[医药卫生—消化系统]