白头翁醇提物对大鼠结肠炎肠黏膜上皮细胞紧密连接蛋白的保护作用  

Protective effects of radix pulsatillae alcohol extract on intestinal epithelial tight junctions in rats with trinitrobenzene sulfonic acidinduced colitis

作　　者：张文远[1] 姜伟炜[1,2] 

机构地区：[1]四川省医学科学院.四川省人民医院消化科,四川省成都市610072 [2]贵州省遵义医学院,贵州省遵义市563003

出　　处：《世界华人消化杂志》2009年第30期3134-3139,共6页World Chinese Journal of Digestology

基　　金：四川省卫生厅基金资助项目;No.070054~~

摘　　要：目的:探讨白头翁醇提物对三硝基苯磺酸诱导大鼠结肠炎肠黏膜上皮细胞紧密连接蛋白的调控,进一步阐明白头翁醇提物对大鼠实验性结肠炎的肠黏膜屏障的保护作用.方法:建立TNBS诱导大鼠结肠炎模型.实验分为正常组、模型组、白头翁醇提物治疗组和双歧杆菌嗜酸乳杆菌肠球菌三联活菌(金双歧)组.进行疾病活动指数(DAI)和组织学损伤评分,用ELISA法测定结肠组织TNF-α、IL-10和血清内毒素,采用免疫组织学染色检测紧密连接(tightjunction,TJ)相关蛋白occludin的分布.结果:TNBS诱导大鼠结肠炎后,DAI和组织学损伤评分增高,结肠组织TNF-α水平升高、IL-10水平降低和血清内毒素水平升高,而经白头翁醇提物和双歧杆菌嗜酸乳杆菌肠球菌三联活菌处理后,DAI和组织学损伤评分有明显下降(6.50±1.27,5.90±1.67vs9.20±1.75;5.00±1.05,4.80±1.25vs7.10±0.99,均P<0.05),结肠组织TNF-α水平降低(521.24±109.37ng/L,503.98±126.63ng/Lvs657.54±149.60ng/L,均P<0.05)、血清内毒素水平降低(0.148±0.093EU/mL,0.153±0.106EU/mLvs0.213±0.023EU/mL,均P<0.05)和IL-10水平升高(92.19±30.09ng/L,95.57±27.71ng/Lvs42.92±23.74ng/L,均P<0.05);TNBS诱导大鼠结肠炎后,TJ结构遭到破坏,TJ相关蛋白的表达减少,而白头翁醇提物和双歧杆菌嗜酸乳杆菌肠球菌三联活菌(金双歧)处理后可使TNBS引起的TJ结构受损减轻,相关蛋白的表达增多.结论:白头翁醇提物可以对TNBS诱导大鼠结肠炎肠黏膜屏障具有明显的保护作用,其机制可能是通过调节肠道微生态、上调肠上皮细胞紧密连接蛋白occludin的表达、降低结肠组织TNF-α含量、提高IL-10水平,从而抑制内毒素通过紧密连接进入体循环.AIM： To investigate the protective effects of radix pulsatillae alcohol extract on intestinal epithelial tight junctions in rats with trinitrobenzene sulfonic acid （TNBS）-induced colitis and explore potential mechanisms involved. METHODS： Acute colitis was induced in rats using TNBS enema. Thirty-eight rats were randomized into four groups： normal control group （n = 8）, model control group （n = 10）, radixpulsatillae alcohol extract treatment group （n = 10） and golden bifid （live combined bifidobacterium, lactobacillus and streptococcus thermophilus tablets） treatment group （n = 10）. Diarrhea and bloody stool were observed. Colonic damage was evaluated histologically. The levels of tumor necrosis factor-α （TNF-α） and interleukin-10 （IL-10） in colonic tissue and endotoxin in blood were determined by enzyme-linked immunosorbent assay （ELISA）. The expression of occludin in intestinal mucosal epithelial cells was detected by immunohistochemistry. RESULTS： After acute colitis was induced in rats using TNBS enema, disease activity index and colonic damage score increased. However, treatment with radix pulsatillae alcohol extract and golden bifid significantly decreased disease activity index （6.50 ± 1.27 and 5.90 ± 1.67 vs 9.20 ± 1.75, respectively; both P〈0.05）, colonic damage score （5.00 ± 1.05 and 4.80 ± 1.25 vs 7.10 ± 0.99, respectively; both P〈0.05）, colonic TNF-α level （521.24 ± 109.37 ng/L and 503.98 ± 126.63 ng/L vs 657.54 ± 149.60 ng/L, respectively; both P〈0.05） and blood endotoxin level （0.148 ± 0.093 EU/mL and 0.153 ± 0.106 EU/mL vs 0.213 ± 0.023 EU/mL, respectively; both P〈0.05）, and remarkably increased colonic IL-10 content （92.19 ± 30.09 ng/L and 95.57 ± 27.71 ng/L vs 42.92 ± 23.74 ng/L, respectively; both P〈0.05） in rats with TNBS-induced colitis. In the model control group, the structure of tight junctions was destroyed and the expression of occludin was downregulated in colonic mucosal epithelium. However, trea

关 键 词：肿瘤坏死因子α 白介素10 内毒素 紧密连接蛋白 肠道黏膜屏障 

分 类 号：R285.5[医药卫生—中药学]