摘要
目的研究TNF-α抗体对葡聚糖硫酸钠（DSS）诱导的实验性结肠炎小鼠肠黏膜通透性的影响及作用机制。方法将18只C57BL／6J小鼠均分为健康对照组、模型对照组和TNF-α抗体组，其中模型对照组和TNF-α抗体组小鼠喂饲5％DSS溶液7d。TNF-α抗体组分别于造模第1、4天予TNF—α抗体（5mg／kg）腹腔注射给药，健康对照组和模型对照组则以等量0．9％氯化钠溶液代替。造模7d后处死小鼠，期间每日进行疾病活动指数（DAI）评分。分别采用偶氮蓝法和异硫氰酸荧光素标记葡聚糖（FITC—D）法检测小肠黏膜通透性。取结肠肠段，光学显微镜下观察并行病理组织学评分。取部分回肠肠段，电子显微镜下观察。制备10％结肠和小肠黏膜匀浆，使用相应试剂盒测定结肠黏膜髓过氧化物酶（MP0）活性、TNF-α含量和小肠黏膜肌球蛋白轻链激酶（MLCK）含量。Western印迹法检测小肠黏膜MLCK表达水平。统计学处理采用组间单因素方差分析。结果与健康对照组比较，模型对照组小鼠DAI逐日增高。与模型对照组比较，TNF-α抗体组DAI获改善。模型对照组小鼠回肠上皮细胞胞间连接复合体缩短、变宽，细胞间隙扩大。TNF-α抗体组小鼠回肠上皮细胞连接结构紧密程度较高。模型对照组小鼠病理评分和结肠黏膜MPO活性、TNF-α含量均高于健康对照组（P值分别-0．008、0．006、0．001），TNpd抗体组则均低于模型对照组（P值分别-0．004、0．008、0．005），三组间比较F值分别-131．98、218．28、58．93。模型对照组小鼠肠壁中偶氮蓝和血清中FITC-D含量均高于健康对照组（P值分别-0．003和0．010），TNF-α抗体组则均低于模型对照组（P值分别-0．001和0．009），三组间比较F值分别-69．36和17．96。模型对照组小鼠小肠黏膜中MLCK含量[（71．10±7．52）ng／g]高于健康对照组[（18．56±9．92）ng／g，P〈0．01]，TNF
Objective To investigate the effect and mechanism of anti-tumor necrosis factor （TNF）-α on the intestinal mucosal permeability in dextran sulfate sodium （DSS） induced colitis mice. Methods Eighteen C57BL/6J mice were evenly divided into healthy control group, model group and anti-TNF-α treated group. The mice of model group and anti-TNF-α treated group were fed with 5 DSS solution for 7 days. The mice of anti-TNF-α treated group were injected anti-TNF-α （5 mg/kg） intraperitoneally on the first and fourth day; control group and model group were substituted with equal volume saline injection. The miee were sacrificed at 7 days after modeling. The disease activity index （DAD score was evaluated everyday. The intestinal permeability was examined by Evan^s blue （EB） method and FITC-dextran （FITC-D） method. The colon tissue was collected for observation under microscope and histological index （HI）. The small intestinal tissues were examined under electron microscope. The 10H homogenate of colon and intestinal mucosa was prepared, the aetivity of myeloperoxidase （MPO）, the content of TNF-α and epithelial myosin light chain kinase （MLCK） concentration were determined with kits respectively. The expression of MLCK in intestinal mucosa was tested by Western blot assay. Single factor of variance between groups were analyzed. Results Compared with control group, the DAI of model group increased daily. Compared with model group, the DAI of anti-TNF-α treated group improved. In model group, mice intestinal epithelial ceils junetional complex shortened and widened and the cell gap expanded. In anti-TNF-α treated group, the connection structure of mice intestinal epithelial cells was tighter. The aetivity of HI and MPO and the content of TNF-α of model group were higher than those of control group （P= 0. 008, 0. 006 and 0. 001, respectively）, all of those of anti-TNF-a treated group were lower than those of model group （P=0. 004, 0. 008 and 0. 005, respectively）. The F value of
出处
《中华消化杂志》
CAS
CSCD
北大核心
2012年第6期389-394,共6页
Chinese Journal of Digestion
关键词
肿瘤坏死因子α
抗体
结肠炎
肠黏膜
细胞膜通透性
肌球蛋白轻链激酶
Tumor necrosis factor-alpha
Antibodies
Colitis
Intestinal mucosa, Cell membrane permeability
Myosin-light-chain kinase
