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 investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） w...AIM： To investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） were used in this study. A group of rats were administered orally the probiotic L. salivarius ssp. salivarius （5 × 10^8 CFU suspended in 0.5 mL of skimmed milk） daily for 3 wk. Two additional groups were used for reference, a non-colitic and a control colitic without probiotic treatment, which received orally the vehicle used to administer the probiotic. Two weeks after starting the experiment, the rats were rendered colitic by intracolonic administration of 10 mg of TNBS dissolved in 0.25 mL of 500 mL/L ethanol. One week after colitis induction, all animals were killed and colonic damage was evaluated both histologically and biochemically. The biochemical studies performed in colonic homogenates include determination of myeloperoxidase （MPO） activity, glutathione （GSH） content, leukotriene B4 （LTB4） and tumor necrosis factor α （TNF-α） levels, as well as inducible nitric oxide synthase （iNOS） expression. In addition, the luminal contents obtained from colonic samples were used for microbiological studies, in order to determine Lactobacilli and Bifidobacteria counts. RESULTS： Treatment of colitic rats with L. salivarius ssp. salivarius resulted in amelioration of the inflammatory response in colitic rats, when compared with the corresponding control group without probiotic treatment. This antiinflammatory effect was evidenced macroscopically by a significant reduction in the extent of colonic necrosis and/or inflammation induced by the administration of TNBS/ethanol （2.3±0.4 cm vs3.4±0.3 cm in control group, P〈0.01） and histologically by improvement of the colonic architecture associated with a reduction in the neutrophil infiltrate in comparison with non-treated colitic rats. The latter was confirmed biochemically by a significant reduction of colonic MPO activity （105.3±26.0 U/g vs 180.6±21.9 U/g, P〈0.05）, a marker of neutrophil infiltration. The beneficial effect was associated with an increase of the colonic GSH content （1 252±42 nmol/g vs i 087±51 nmol/g, P〈0.05）, which is depleted in colitic rats, as a consequence of the oxidative stress induced by the inflammatory process. In addition, the treatment of colitic rats with L. salivarius resulted in a significant reduction of colonic TNF-（~ levels （509.4±68.2 pg/g vs782.9±60.1 pg/g, P〈0.01） and in a lower colonic iNOS expression, when compared to TNBS control animals without probiotic administration. Finally, treated colitic rats showed higher counts of Lactobacilli species in colonic contents than control colitic rats, whereas no differences were observed in Bifidobacteria counts. CONCLUSION： Administration of the probiotic L. salivarius ssp. salivarius CECT5713 facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with amelioration of the production of some of the mediators involved in the inflammatory response in the intestine, such as cytokines, including TNF-α and NO. This beneficial effect could be ascribed to its effect on the altered immune response that occurs in this inflammatory condition.展开更多
AIM： To evaluate the role of nitric oxide （NO） in the motor disorders of the dilated uninflamed mid-colon （DUMC） from trinitrobenzene sulfonic acid （TNBS）-induced acute distal colitis in rats. METHODS： Colitis...AIM： To evaluate the role of nitric oxide （NO） in the motor disorders of the dilated uninflamed mid-colon （DUMC） from trinitrobenzene sulfonic acid （TNBS）-induced acute distal colitis in rats. METHODS： Colitis was induced in male Sprague-Dawley rats by a single intracolonic administration of TNBS. Control rats received an enema of 0.9% saline. The rats were killed 48 h after TNBS or saline administration. Macroscopic and histologic lesions of the colon were evaluated. Myeloperoxidase （MPO） and nitric oxide synthase （NOS） activity were measured on the colonic tissue. In TNBS rats, we evaluated spontaneous and evoked contractile activity in circular muscle strips derived from DUMC in comparison to the same colonic segment of control rats, both in the presence and in the absence of a non-selective NOS isoforms inhibitor N-nitro-L- arginine （L-NNA）. Pharmacological characterization of electric field stimulation （EFS）-evoked contractile responses was also performed. RESULTS： In TNBS rats, the distal colon showed severe histological lesions and a high MPO activity, while the DUMC exhibited normal histology and MPO activity. Constitutive NOS activity was similar in TNBS and control rats, whereas inducible NOS activity was significantly increased only in the injured distal colon of TNBS rats. Isometrically recorded mechanical activity of circular muscle strips from DUMC of TNBS rats showed a marked reduction of the force and frequency of spontaneous contractions compared to controls, as well as of the contractile responses to a contracting stimulus. In the presence of L-NNA, the contractile activity and responses displayed a significantly greater enhancement compared to controls. The pharmacological characterization of EFS contractile responses showed that a cooperative-like interaction between cholinergic muscarinic and tachyldnergic neurokinin 1 and 2 receptors mediated transmission in DUMC of TNBS rats vs a simple additive interaction in controls. CONCLUSION： The results of this study show that, during TNBS-induced acute distal colitis, circular muscle intrinsic contractile mechanisms and possible enteric neural excitatory activity are inhibited in the distended uninflamed mid-colon. Suppression of NO synthesis markedly improves spontaneous and evokes muscle contractions, in spite of any evident change in local NO activity. 2005 The WJG Press and Elsevier Inc. All rights reserved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。
基金Supported by the Spanish Ministry of Science and Technology, No. SAF2002-02592 and by Institute de Salud 'Carlos Ⅲ', No. PI021732, with Funds from the European Union, and by Junta de Andalucia (CTS 164) Monica Comalada is a recipient of Juan de la Cierva Program from Spanish Ministry of Science and Technology. Laura Peran is a Recipient From Puleva Foundation Spain
文摘AIM： To investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） were used in this study. A group of rats were administered orally the probiotic L. salivarius ssp. salivarius （5 × 10^8 CFU suspended in 0.5 mL of skimmed milk） daily for 3 wk. Two additional groups were used for reference, a non-colitic and a control colitic without probiotic treatment, which received orally the vehicle used to administer the probiotic. Two weeks after starting the experiment, the rats were rendered colitic by intracolonic administration of 10 mg of TNBS dissolved in 0.25 mL of 500 mL/L ethanol. One week after colitis induction, all animals were killed and colonic damage was evaluated both histologically and biochemically. The biochemical studies performed in colonic homogenates include determination of myeloperoxidase （MPO） activity, glutathione （GSH） content, leukotriene B4 （LTB4） and tumor necrosis factor α （TNF-α） levels, as well as inducible nitric oxide synthase （iNOS） expression. In addition, the luminal contents obtained from colonic samples were used for microbiological studies, in order to determine Lactobacilli and Bifidobacteria counts. RESULTS： Treatment of colitic rats with L. salivarius ssp. salivarius resulted in amelioration of the inflammatory response in colitic rats, when compared with the corresponding control group without probiotic treatment. This antiinflammatory effect was evidenced macroscopically by a significant reduction in the extent of colonic necrosis and/or inflammation induced by the administration of TNBS/ethanol （2.3±0.4 cm vs3.4±0.3 cm in control group, P〈0.01） and histologically by improvement of the colonic architecture associated with a reduction in the neutrophil infiltrate in comparison with non-treated colitic rats. The latter was confirmed biochemically by a significant reduction of colonic MPO activity （105.3±26.0 U/g vs 180.6±21.9 U/g, P〈0.05）, a marker of neutrophil infiltration. The beneficial effect was associated with an increase of the colonic GSH content （1 252±42 nmol/g vs i 087±51 nmol/g, P〈0.05）, which is depleted in colitic rats, as a consequence of the oxidative stress induced by the inflammatory process. In addition, the treatment of colitic rats with L. salivarius resulted in a significant reduction of colonic TNF-（~ levels （509.4±68.2 pg/g vs782.9±60.1 pg/g, P〈0.01） and in a lower colonic iNOS expression, when compared to TNBS control animals without probiotic administration. Finally, treated colitic rats showed higher counts of Lactobacilli species in colonic contents than control colitic rats, whereas no differences were observed in Bifidobacteria counts. CONCLUSION： Administration of the probiotic L. salivarius ssp. salivarius CECT5713 facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with amelioration of the production of some of the mediators involved in the inflammatory response in the intestine, such as cytokines, including TNF-α and NO. This beneficial effect could be ascribed to its effect on the altered immune response that occurs in this inflammatory condition.
文摘AIM： To evaluate the role of nitric oxide （NO） in the motor disorders of the dilated uninflamed mid-colon （DUMC） from trinitrobenzene sulfonic acid （TNBS）-induced acute distal colitis in rats. METHODS： Colitis was induced in male Sprague-Dawley rats by a single intracolonic administration of TNBS. Control rats received an enema of 0.9% saline. The rats were killed 48 h after TNBS or saline administration. Macroscopic and histologic lesions of the colon were evaluated. Myeloperoxidase （MPO） and nitric oxide synthase （NOS） activity were measured on the colonic tissue. In TNBS rats, we evaluated spontaneous and evoked contractile activity in circular muscle strips derived from DUMC in comparison to the same colonic segment of control rats, both in the presence and in the absence of a non-selective NOS isoforms inhibitor N-nitro-L- arginine （L-NNA）. Pharmacological characterization of electric field stimulation （EFS）-evoked contractile responses was also performed. RESULTS： In TNBS rats, the distal colon showed severe histological lesions and a high MPO activity, while the DUMC exhibited normal histology and MPO activity. Constitutive NOS activity was similar in TNBS and control rats, whereas inducible NOS activity was significantly increased only in the injured distal colon of TNBS rats. Isometrically recorded mechanical activity of circular muscle strips from DUMC of TNBS rats showed a marked reduction of the force and frequency of spontaneous contractions compared to controls, as well as of the contractile responses to a contracting stimulus. In the presence of L-NNA, the contractile activity and responses displayed a significantly greater enhancement compared to controls. The pharmacological characterization of EFS contractile responses showed that a cooperative-like interaction between cholinergic muscarinic and tachyldnergic neurokinin 1 and 2 receptors mediated transmission in DUMC of TNBS rats vs a simple additive interaction in controls. CONCLUSION： The results of this study show that, during TNBS-induced acute distal colitis, circular muscle intrinsic contractile mechanisms and possible enteric neural excitatory activity are inhibited in the distended uninflamed mid-colon. Suppression of NO synthesis markedly improves spontaneous and evokes muscle contractions, in spite of any evident change in local NO activity. 2005 The WJG Press and Elsevier Inc. All rights reserved
