目的:研究三叶因子3(trefoil factor 3,TFF3)基因在三硝基苯磺酸(TNBS)诱发的大鼠结肠炎肠黏膜表达的变化,探讨其在结肠炎发生发展中的作用. 方法:采用TNBS/乙醇灌肠制作大鼠结肠炎模型(n=15), 观察大鼠腹泻、便血情况,评价疾病活动性指...目的:研究三叶因子3(trefoil factor 3,TFF3)基因在三硝基苯磺酸(TNBS)诱发的大鼠结肠炎肠黏膜表达的变化,探讨其在结肠炎发生发展中的作用. 方法:采用TNBS/乙醇灌肠制作大鼠结肠炎模型(n=15), 观察大鼠腹泻、便血情况,评价疾病活动性指数(DAI)并记录体重改变,分别于制模后1,7及14d处死大鼠.生理盐水灌肠作为正常对照组.大体及镜下观察肠黏膜损伤及组织学变化,并进行评分.生化法检测MPO活性.逆转录-多聚酶链反应(RT-PCR)检测TFF3基因表达. 结果:结肠炎组1,7及14d时间点DAI(3.8±0.5 vs 0, 3.3±0.5 vs 0,2.3±0.5 vs 0)、体质量(-3.2±0.7 vs 4.7±2.2, -7.2±2.0 vs 10.9±0.2,2.9±0.4 vs 30.5±2.9)、大体评分(7.2±0.9 vs 0,6.2±1.3 vs 0,4.6±0.5 vs 0)和组织学评分(8.2±1.2 vs 0,10.4±0.5 vs 0,8.4±0.5 vs 0)以及MPO活性(1745±55 vs 303±21,1789±77 vs 315±20,1736±127 vs 313±35)较正常对照组均存在显著差异(P<0.05).TFF3在结肠组织炎症的各个时间点均有表达,致炎后1d组较正常对照组减少,7和14 d组明显升高(0.63±0.05 vs 0.72±0.02, 0.94±0.19 vs 0.72±0.02.1.25±0.74 vs 0.72±0.02.P<0.05). 结论:TNBS诱发的大鼠结肠炎有TFF3基因的表达,提示TFF3在结肠炎黏膜损伤修复中发挥作用.展开更多
AIM: To evaluated the therapeutic and prophylactic effect of thalidomide on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Thalidomide has been reported to downregulate the expression of tumor necrosis ...AIM: To evaluated the therapeutic and prophylactic effect of thalidomide on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Thalidomide has been reported to downregulate the expression of tumor necrosis factor a (TNF-α), IL-12, and vascular endothelial growth factor (VEGF), hallmarks of intestinal inflammation in Crohn's disease (CD). METHODS: Male Wistar rats were divided in five groups of ten animals each. Four groups received a rectal infusion of TNBS in ethanol. The first group was sacrificed 7 d after colitis induction. The second and third groups received either thalidomide or placebo by gavage and were sacrificed at 14 d. The fourth group received thalidomide 6 h before TNBS administration, and was sacrificed 7 d after induction. The fifth group acted as the control group and colitis was not induced. Histological inflammatory scores of the colon were performed and lamina propria CD4+ T cells, macrophages, and VEGF+ cells were detected by immunohistochemistry. TNF-a and IL-12 were quantified in the supernatant of organ cultures by ELISA. RESULTS: Significant reduction in the inflammatory score and in the percentage of VEGF+ cells was observed in the group treated with thalidomide compared with animals not treated with thalidomide. Both TNF-αand IL-12 levels were significantly reduced among TNBS induced colitis animals treated with thalidomide compared with animals that did not receive thalidomide. TNF-αlevels were also significantly reduced among the animals receiving thalidomide prophylaxis compared with untreated animals with TNBS-induced colitis. Intestinal levels of TNF-αand IL-12 were significantly correlated with the inflammatory score and the number of VEGF+ cells. CONCLUSION: Thalidomide significantly attenuates TNBS-induced colitis by inhibiting the intestinal production of TNF-α, IL-12, and VEGF. This effect may support the use of thalidomide as an alternate approach in selected patients with CD.展开更多
目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂罗格列酮(rosiglitazone)对2,4,6-三硝基苯磺酸(TNBS诱导的结肠炎小鼠的治疗作用及可能机制。方法选取雄性BALB/c小鼠20只建立TNBS结肠炎模型,造模成功后随机分为罗格列酮处理组及...目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂罗格列酮(rosiglitazone)对2,4,6-三硝基苯磺酸(TNBS诱导的结肠炎小鼠的治疗作用及可能机制。方法选取雄性BALB/c小鼠20只建立TNBS结肠炎模型,造模成功后随机分为罗格列酮处理组及模型组,每组10只。罗格列酮处理组给予罗格列酮灌胃(0. 2 m L,[20 mg/(kg·d)]),模型组给予生理盐水(0. 2 m L/d)灌胃。处理6周后,处死所有小鼠。采用炎症性肠病疾病活动度指数(DAI)及HE染色结合Spencer结肠炎组织学评分评估两组小鼠肠道炎症程度及组织学改变,ELISA检测肠黏膜白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-10水平及肠系膜脂肪组织IL-6、单核细胞趋化蛋白1(MCP-1)水平;脂肪组织HE染色后200倍光镜下拍照依据比例尺计算脂肪细胞直径;免疫组织化学染色法检测小鼠肠系膜脂肪组织F4/80阳性巨噬细胞浸润数量、脂肪细胞成熟标志物外周蛋白(peripherin)、脂连蛋白(adiponectin)及瘦素(leptin)水平。Western blot法检测小鼠肠系膜脂肪组织核因子κB(NF-κB)和信号通路中的NF-κBp65、磷酸化的NF-κBp65(p-NF-κBp65)、磷酸化的NF-κB抑制蛋白(p-IκB)及磷酸化的IκB激酶(p-IKK)的蛋白水平。结果罗格列酮处理后第5及6周,小鼠DAI评分显著低于模型组。同时,处理组小鼠肠黏膜IL-1β及TNF-α水平显著低于模型组,而IL-10水平显著高于模型组。与模型组小鼠相比,罗格列酮处理组小鼠肠系膜脂肪细胞直径及脂肪细胞成熟标志物perilipin水平均显著高于模型组。同时,罗格列酮处理组小鼠肠系膜脂肪组织巨噬细胞浸润数量及炎症介质IL-6、MCP-1水平均显著低于模型组。罗格列酮治疗显著促进处理组小鼠肠系膜脂肪细胞adiponectin的表达,而抑制leptin的水平。罗格列酮处理组小鼠肠系膜脂肪组织NF-κBp65、p-NF-κBp65、p-IKK及p-IκB蛋白水平均显著低于模型组。结论罗格列酮可显著抑制TNBS模型小鼠肠道炎症,可能与抑制肠系膜脂肪组织NF-κBp65通路有关。展开更多
文摘目的:研究三叶因子3(trefoil factor 3,TFF3)基因在三硝基苯磺酸(TNBS)诱发的大鼠结肠炎肠黏膜表达的变化,探讨其在结肠炎发生发展中的作用. 方法:采用TNBS/乙醇灌肠制作大鼠结肠炎模型(n=15), 观察大鼠腹泻、便血情况,评价疾病活动性指数(DAI)并记录体重改变,分别于制模后1,7及14d处死大鼠.生理盐水灌肠作为正常对照组.大体及镜下观察肠黏膜损伤及组织学变化,并进行评分.生化法检测MPO活性.逆转录-多聚酶链反应(RT-PCR)检测TFF3基因表达. 结果:结肠炎组1,7及14d时间点DAI(3.8±0.5 vs 0, 3.3±0.5 vs 0,2.3±0.5 vs 0)、体质量(-3.2±0.7 vs 4.7±2.2, -7.2±2.0 vs 10.9±0.2,2.9±0.4 vs 30.5±2.9)、大体评分(7.2±0.9 vs 0,6.2±1.3 vs 0,4.6±0.5 vs 0)和组织学评分(8.2±1.2 vs 0,10.4±0.5 vs 0,8.4±0.5 vs 0)以及MPO活性(1745±55 vs 303±21,1789±77 vs 315±20,1736±127 vs 313±35)较正常对照组均存在显著差异(P<0.05).TFF3在结肠组织炎症的各个时间点均有表达,致炎后1d组较正常对照组减少,7和14 d组明显升高(0.63±0.05 vs 0.72±0.02, 0.94±0.19 vs 0.72±0.02.1.25±0.74 vs 0.72±0.02.P<0.05). 结论:TNBS诱发的大鼠结肠炎有TFF3基因的表达,提示TFF3在结肠炎黏膜损伤修复中发挥作用.
文摘目的应用不同浓度TNBS与乙醇的混合液灌肠制备克罗恩病大鼠模型,对比结果进行评价研究,寻求制备克罗恩病大鼠最佳的TNBS与乙醇浓度,为克罗恩病的基础研究提供可靠的动物模型。方法 SPF级大鼠60只,采用分层随机法分为5组;分别予以体积比为1∶1的双蒸水和无水乙醇混合液、2∶1的TNBS和50%乙醇混合液、1∶1的TNBS和50%乙醇混合液、2∶1的TNBS和无水乙醇混合液、1∶1的TNBS和无水乙醇混合液按0.6 m L/只(100~150 mg/kg)灌肠,比较造模后大鼠不同的一般行为学、粪便及含水量的测定、体重、组织病理学等指标的差异。结果与对照组比较,模型组大鼠出现精神状态差、粪便含水量高、结肠肉眼、镜下观均有不同损伤等现象,提示制备出不同病变程度的克罗恩病动物模型。结论制备克罗恩病大鼠模型所需的最佳TNBS与乙醇浓度比例为TNBS:无水乙醇=2∶1。
文摘AIM: To evaluated the therapeutic and prophylactic effect of thalidomide on 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Thalidomide has been reported to downregulate the expression of tumor necrosis factor a (TNF-α), IL-12, and vascular endothelial growth factor (VEGF), hallmarks of intestinal inflammation in Crohn's disease (CD). METHODS: Male Wistar rats were divided in five groups of ten animals each. Four groups received a rectal infusion of TNBS in ethanol. The first group was sacrificed 7 d after colitis induction. The second and third groups received either thalidomide or placebo by gavage and were sacrificed at 14 d. The fourth group received thalidomide 6 h before TNBS administration, and was sacrificed 7 d after induction. The fifth group acted as the control group and colitis was not induced. Histological inflammatory scores of the colon were performed and lamina propria CD4+ T cells, macrophages, and VEGF+ cells were detected by immunohistochemistry. TNF-a and IL-12 were quantified in the supernatant of organ cultures by ELISA. RESULTS: Significant reduction in the inflammatory score and in the percentage of VEGF+ cells was observed in the group treated with thalidomide compared with animals not treated with thalidomide. Both TNF-αand IL-12 levels were significantly reduced among TNBS induced colitis animals treated with thalidomide compared with animals that did not receive thalidomide. TNF-αlevels were also significantly reduced among the animals receiving thalidomide prophylaxis compared with untreated animals with TNBS-induced colitis. Intestinal levels of TNF-αand IL-12 were significantly correlated with the inflammatory score and the number of VEGF+ cells. CONCLUSION: Thalidomide significantly attenuates TNBS-induced colitis by inhibiting the intestinal production of TNF-α, IL-12, and VEGF. This effect may support the use of thalidomide as an alternate approach in selected patients with CD.
文摘目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂罗格列酮(rosiglitazone)对2,4,6-三硝基苯磺酸(TNBS诱导的结肠炎小鼠的治疗作用及可能机制。方法选取雄性BALB/c小鼠20只建立TNBS结肠炎模型,造模成功后随机分为罗格列酮处理组及模型组,每组10只。罗格列酮处理组给予罗格列酮灌胃(0. 2 m L,[20 mg/(kg·d)]),模型组给予生理盐水(0. 2 m L/d)灌胃。处理6周后,处死所有小鼠。采用炎症性肠病疾病活动度指数(DAI)及HE染色结合Spencer结肠炎组织学评分评估两组小鼠肠道炎症程度及组织学改变,ELISA检测肠黏膜白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-10水平及肠系膜脂肪组织IL-6、单核细胞趋化蛋白1(MCP-1)水平;脂肪组织HE染色后200倍光镜下拍照依据比例尺计算脂肪细胞直径;免疫组织化学染色法检测小鼠肠系膜脂肪组织F4/80阳性巨噬细胞浸润数量、脂肪细胞成熟标志物外周蛋白(peripherin)、脂连蛋白(adiponectin)及瘦素(leptin)水平。Western blot法检测小鼠肠系膜脂肪组织核因子κB(NF-κB)和信号通路中的NF-κBp65、磷酸化的NF-κBp65(p-NF-κBp65)、磷酸化的NF-κB抑制蛋白(p-IκB)及磷酸化的IκB激酶(p-IKK)的蛋白水平。结果罗格列酮处理后第5及6周,小鼠DAI评分显著低于模型组。同时,处理组小鼠肠黏膜IL-1β及TNF-α水平显著低于模型组,而IL-10水平显著高于模型组。与模型组小鼠相比,罗格列酮处理组小鼠肠系膜脂肪细胞直径及脂肪细胞成熟标志物perilipin水平均显著高于模型组。同时,罗格列酮处理组小鼠肠系膜脂肪组织巨噬细胞浸润数量及炎症介质IL-6、MCP-1水平均显著低于模型组。罗格列酮治疗显著促进处理组小鼠肠系膜脂肪细胞adiponectin的表达,而抑制leptin的水平。罗格列酮处理组小鼠肠系膜脂肪组织NF-κBp65、p-NF-κBp65、p-IKK及p-IκB蛋白水平均显著低于模型组。结论罗格列酮可显著抑制TNBS模型小鼠肠道炎症,可能与抑制肠系膜脂肪组织NF-κBp65通路有关。
