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目的探讨细菌脂多糖免疫对炎症性肠病小鼠肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达的影响及其作用机制。方法 BALB/C雌性小鼠10只随机分为实验组5只和对照组5只,实验组腹腔内注射1.0mg/kg细菌脂多糖和免疫佐剂的混合物,对照组腹腔内注射免疫佐剂1.0mg/kg,2组小鼠均每周注射1次,注射4次。末次注射1周后均给予150mg/kg的三硝基苯磺酸乙醇溶液灌肠诱导炎症性肠病。观察2组小鼠一般状态、体质量改变情况、粪便性状,检测粪便隐血;灌肠1周后处死小鼠,观察结肠组织学改变,采用ELISA法检测结肠组织TNF-α表达情况,采用Western blot法检测结肠组织核因子(nuclear factor,NF)-κB表达情况。结果 2组小鼠灌肠后第2天均出现懒动、厌食、皮毛凌乱、腹泻,第3天出现血便、体质量下降,实验组上述表现较对照组轻,恢复快;实验组小鼠第3天体质量下降克数((1.60±0.38)g)、粪便隐血阳性持续时间((2.40±0.54)d)、结肠组织学评分((3.72±2.45)分)、结肠组织TNF-α表达水平((20.16±3.21)ng/L)和NF-κB吸光度值((18.43±5.36)×10-3)均明显低于对照组((2.80±0.45)g、(3.60±0.46)d、(6.39±1.72)分、(32.35±2.80)ng/L、(25.80±3.79)×10-3)(P<0.05)。结论细菌脂多糖免疫可降低炎症性肠病小鼠结肠组织TNF-α和NF-κB表达水平,减轻结肠组织病理损伤,机体对细菌脂多糖反应异常可能是炎症性肠病的发病因素之一。
Objective To investigate the effect of bacterial lipopolysaccharide immunization on the expression of tumor necrosis factor-α (TNF-α) and its mechanism in inflammatory bowel disease mice. Methods Ten BALB / C female mice were randomly divided into experimental group (n = 5) and control group (n = 5). The experimental group was intraperitoneally injected with a mixture of 1.0 mg / kg bacterial lipopolysaccharide and immune adjuvant. The control group received intraperitoneal injection of adjuvant mg / kg, two groups of mice were injected once a week and injected four times. One week after the last injection were given 150mg / kg of trinitrobenzene sulfonic acid ethanol enema induced inflammatory bowel disease. The mice were sacrificed one week after the enema. The histological changes of the colon were observed. The expression of TNF-α in colonic tissue was detected by ELISA. Western blot was used to detect the expression of TNF- The expression of nuclear factor (NF) -κB in colonic tissue was detected. Results In the 2nd day after the enema, lazyness, anorexia, fur mess and diarrhea appeared on the 2nd day after enema in mice. The bloody stool appeared on the 3rd day and the body weight decreased. The performance of the above two groups was lighter and recovered faster than the control group. (1.60 ± 0.38) g, the duration of concealed fecal occult blood test (2.40 ± 0.54 d), colon histological score (3.72 ± 2.45), the level of TNF-α in colonic tissue (20.16 ± 3.21) ng / L and NF-κB absorbance (18.43 ± 5.36 × 10-3) were significantly lower than those in the control group (2.80 ± 0.45g, 3.60 ± 0.46d, 6.39 ± 1.72 ), (32.35 ± 2.80) ng / L, (25.80 ± 3.79) × 10-3) (P <0.05). Conclusions Bacterial lipopolysaccharide immunosuppression can reduce the expression of TNF-α and NF-κB in colonic tissue of mice with inflammatory bowel disease and reduce the pathological damage of colonic tissue. The abnormal reaction of bacterial to lipopolysaccharide may be one of the pathogenesis of inflammatory bowel disease.