吉法酯对三硝基苯磺酸诱导大鼠实验性结肠炎的作用

目的 观察吉法酯对三硝基苯磺酸(TNBS)诱导的大鼠实验性结肠炎髓过氧化物酶(MPO)、环氧合酶(COX)-1及COX-2表达的影响,探讨吉法酯对溃疡性结肠炎的治疗作用.方法 选用雌性健康SD大鼠40只,均分为A、B、C、D组.A、B、C三组大鼠采用TNBS/乙醇灌肠制作大鼠结肠炎模型.造模后第2天,A组每天给予0.9%氯化钠溶液1 ml灌肠;B组每天给予5-氨基水杨酸(5-ASA)1 ml灌肠(100 mg/kg);C组每天给予吉法酯1 ml灌胃.D组为正常对照组.分别于造模后第7天及第14天每组处死5只大鼠,按疾病活动指数(DAI)的评分标准进行大体损伤评分,HE染色进行组织损伤评分.同时取结肠病变部位组织,生化法检测MPO活性,免疫组化法检测COX-1与COX-2的组织表达.结果 与A组比较,B组和C组的DAI评分、大体损伤形态和组织学损伤评分及MPO活性均降低(P<0.05).与A组相比,B、C、D组第7天和第14天COX-1表达水平升高(P<0.05),分别为0.87±0.18和0.93±0.15比1.86±0.51和1.96±0.41,1.73±0.68和1.79±0.6以及1.91±0.34和1.99±0.45;COX-2水平降低(P<0.05),分别为3.50±0.23和3.06±0.27比1.53±0.19和0.73±0.15,1.73±0.94和0.86±0.29,0.24±0.18和0.18±0.16.D组COX-2表达极弱,与B、C两组间差异有统计学意义(P<0.05).结论 吉法酯对TNBS诱导的大鼠结肠炎有较好的治疗作用,其疗效与5-ASA相似,其作用机制可能是降低肠组织中MPO的活性和调节COX-1/COX-2表达.     

徐长卿对三硝基苯磺酸诱导的大鼠结肠炎的作用

目的:探讨徐长卿对2,4,6-三硝基苯磺酸(trinitrobenzenesulfonic acid,TNBS)诱导的大鼠结肠炎的作用.方法:将40只♂SD大鼠随机分为4组:正常组、模型组、徐长卿组和巴柳氮组.除正常组外,其余3组大鼠均以TNBS灌肠造模.灌肠24h后,徐长卿组开始每天给予徐长卿4g/kg;巴柳氮组给予巴柳氮钠1g/kg灌胃治疗10d.每天观察大鼠一般情况,给药结束后,观察大鼠结肠大体损伤及病理,酵素免疫分析法(enzyme-linked immunosorbant assay,ELISA)检测肠组织肿瘤坏死因子(tumor necrosis factor,TNF)-α、白介素(interleukin,IL)-1β及IL-10水平.结果:两治疗组体质量较模型组增加,但差异无统计学意义;两治疗组疾病活动指数(disease activity index,DAI)评分较模型组明显下降(0.70±1.06,0.67±0.71vs2.38±1.51,P<0.05).徐长卿组、巴柳氮组结肠大体损伤及病理评分较模型组显著下降(1.05±0.83,1.06±0.85vs2.94±0.94;1.65±1.67,2.00±1.80vs6.00±1.67,均P<0.01).徐长卿组较模型组TNF-α、IL-1β水平明显降低(P<0.01),IL-10水平无统计学差异.巴柳氮组较模型组TNF-α、IL-1β、IL-10水平均明显降低(P<0.01).结论:徐长卿能有效改善TNBS诱导的大鼠结肠炎,其机制可能与调节细胞因子水平有关.     

大蒜素对三硝基苯磺酸诱导大鼠结肠炎的保护作用及其机制研究

背景：溃疡性结肠炎（UC）是一种病因未明的结直肠炎症,大蒜素对其防治的作用目前尚未有结论。目的：探讨大蒜素对TNBS诱导的大鼠结肠炎的保护作用及其机制。方法：80只大鼠随机分为对照组、TNBS组、大蒜素预防组、大蒜素灌胃组、大蒜素灌肠组、地塞米松组、柳氮磺吡啶组、巴柳氮钠组。以含150mg／kgTNBS的50％乙醇溶液灌肠制备大鼠结肠炎模型。造模2周后处死大鼠。行大体评分和病理学评分,以ELISA法测定血清TNF-α、IL-1β、IL-10、IL4含量,蛋白质印迹法检测NF—κB表达。结果：与对照组相比,TNBS组大体和病理学评分均明显升高（P〈0．05）,体质量明显降低（P〈0．05）,血清TNF-α、IL-1B含量显著升高（P〈0．05）,血清IL-4、IL-10含量显著降低（P〈0．05）,NF-κB表达明显升高（P〈0．05）;给予大蒜素预防或治疗后,上述指标均明显改善（P〈0．05）,但疗效低于地塞米松组、柳氮磺吡啶组、巴柳氮钠组。结论：大蒜素对TNBS诱导的大鼠结肠炎有保护作用,可能是通过调控细胞因子和NF—κB而发挥作用的。     

参青方对三硝基苯磺酸诱导结肠炎大鼠肠道动力学的研究

[目的]探讨参青方对三硝基苯磺酸（TNBS）诱导结肠炎大鼠肠道动力学异常的消炎作用及其调节肠道动力学的机制。[方法]用TNBS复制实验性大鼠结肠炎模型,随机分为参青方高剂量组、参青方低剂量组、美沙拉嗪（5-ASA）组、模型对照（模型）组及正常组,检测结肠肠管平滑肌条收缩频率、幅度。[结果]与正常组相比,造模各组大鼠结肠肠管平滑肌条收缩频率减少,幅度增加（均P〈0．05）;且参青方高、低剂量组及5-ASA组大鼠结肠肠管收缩频率和幅度均比模型组增加（均P〈0．05）。[结论]TNBS诱导结肠炎大鼠出现肠管收缩频率和收缩波幅度异常;参青方能增加收缩频率和收缩波幅度,具有调节肠道动力学的作用。     

2,4,6-三硝基苯磺酸/乙醇法诱导建立溃疡性结肠炎大鼠模型

目的采用2,4,6-三硝基苯磺酸(TNBS)/乙醇法灌肠建立较理想的溃疡性结肠炎(UC)大鼠模型。方法将20只雄性SD大鼠随机分为正常组、模型组。模型组大鼠以TNBS灌肠。每天观察大鼠一般情况,观察大鼠结肠大体损伤及病理。结果模型组大鼠精神萎靡,少动,毛发杂乱无光泽,饮食量减少,出现不同程度的腹泻及便血;肉眼见肠道粘连,肠管积气积粪,肠壁增厚、皱褶消失,肠黏膜广泛充血水肿并可见明显的溃疡灶;病理见黏膜层、黏膜下层溃疡形成,大量中性粒细胞、淋巴细胞等炎性细胞浸润,腺体破坏,结构紊乱,杯状细胞减少,隐窝结构扭曲,隐窝炎症及脓肿形成。结论采用TNBS/乙醇法建立UC模型为较理想的研究模型。     

三硝基苯磺酸诱导小鼠溃疡性结肠炎模型制备的技术改良

目的:改良2,4,6-三硝基苯磺酸(TNBS)硅胶管灌肠诱导制备小鼠溃疡性结肠炎(UC)模型的方法,提高造模的成功率和模型的稳定性,并探索造模的适宜剂量和时间.方法:选用40只SPF级♂Balb/c小鼠,6-8周龄,随机分为正常对照组、不同浓度TNBS组(37.5mg/kg、75mg/kg、150mg/kg、200mg/k g),每组8只,使用"灌胃针"替代"硅胶管"灌肠,并于灌肠后2d和4d分别处死4只小鼠,观察不同组别小鼠生理状态、结肠组织的损伤及病理学的改变情况.结果:在"灌胃针"造模过程中未发生小鼠死亡现象;对照组小鼠一般情况及结肠黏膜组织无异常改变;小鼠灌肠后出现少食、少动、体质量下降、皮毛光泽度下降、腹泻、便血.不同浓度TNBS造模组随着TNBS剂量的增加,小鼠结肠黏膜组织出现充血、出血、水肿、炎症、溃疡的程度增加.HE染色可见结肠组织水肿、炎症细胞浸润、杯状细胞缺失、溃疡形成的程度逐渐增加.其中TNBS37.5mg/kg、75mg/kg组于造模后2d,以上损伤现象开始缓解,未形成稳定的UC模型;150mg/kg、200mg/kg组持续时间较长,以上损伤现象4d内未见明显缓解,150mg/kg组表现为较典型的UC模型,200mg/kg为重症UC模型.结论:对制造小鼠UC模型进行相关技术改进,使灌肠更加简便,提高造模效率,显著增加了模型的稳定性.     

雷公藤红素对三硝基苯磺酸诱导的大鼠结肠炎的保护作用

背景：传统药物对炎症性肠病疗效不甚理想，寻找新型而有效的药物一直是该领域的研究热点。目的：观察雷公藤红素对三硝基苯磺酸（TNBS）诱导的大鼠结肠炎的保护作用，并初步探讨其可能机制。方法：以TNBS诱导大鼠结肠炎模型。将动物随机分为正常对照组、模型组、助溶剂对照组以及雷公藤红素低剂量组（每天0．5mg／kg）和高剂量组（每天1mg／kg）。以大体和组织学评分评价结肠炎症程度。以免疫组化方法检测结肠组织核因子（NF）-kBp65的表达，以半定量逆转录聚合酶链反应（RT-PCR）检测白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α mRNA的表达。结果：高、低剂量雷公藤红素均能显著改善结肠组织大体和组织学评分，降低NF-kB p65以及IL-1β、TNF-α mRNA的表达。结论：雷公藤红素对TNBS诱导的大鼠结肠炎具有显著保护作用，抑制促炎细胞因子的产生可能是其主要作用机制之一。     

清肠栓调节三硝基苯磺酸诱导结肠炎大鼠结肠黏膜细胞增殖的影响

[目的]从细胞增殖动力学角度探讨清肠栓促进结肠溃疡愈合的作用机制.[方法]制备三硝基苯磺酸(TNBS)诱导结肠炎大鼠.造模3 d,分为清肠栓高剂量组、清肠栓低剂量组、柳氮磺胺吡啶(SASP)组、模型对照组、模型组和正常组.给药7 d后,取大鼠结肠病变部位标本,进行组织学评价,运用AB-PAS染色观察杯状细胞数量及其分泌黏液功能,免疫组化染色法检测增殖细胞核抗原(PCNA)表达.[结果]与清肠栓低剂量组、SASP组、模型对照组和正常组比较,清肠栓高剂量组大鼠结肠黏膜炎症消除和溃疡愈合,杯状细胞数量及黏液增加,溃疡边缘腺体细胞增殖加强,PCNA表达增加(P＜0.05).[结论]清肠栓具有促进结肠炎大鼠结肠黏膜细胞增殖、增加杯状细胞的数量和分泌黏液的水平等作用,能够促进结肠溃疡愈合过程.     

三七提取物对三硝基苯磺酸诱导溃疡性结肠炎大鼠结肠血管生成的研究

[目的]从血管生成角度研究三七提取物促进结肠溃疡愈合的作用机制。[方法]制备三硝基苯磺酸（TNBS）诱导溃疡性结肠炎（UC）大鼠，分为三七提取物低、中、高剂量组，柳氮磺胺吡啶（SASP）组，模型对照组，模型组和正常对照（正常）组。其中模型组于造模3d时处死，其余6组均在给药处理7d（即造模10d）时处死。取大鼠结肠病变部位标本，进行组织学评价；运用免疫组化染色法检测血管内皮生长因子（VEGF）、fms样酪氨酸激酶受体（FLT-1）和含有激酶插入区域的受体（KDR）、低氧诱导因子（HIF-1）蛋白表达；用RT-PCR法检测VEGF、FLT-1和KDR、HIF-1的mRNA表达。[结果]与模型对照组和SASP组比较，三七提取物低、中和高剂量组大鼠结肠黏膜炎症明显消除和溃疡愈合，杯状细胞数量及黏液增加，VEGF、FLT-1和KDR、HIF-1表达显著增加。[结论]血管生成在TNBS诱导大鼠UC形成和愈合过程中起重要作用。用药后VEGF、KDR、FLT-1和HIF-1表达增加。三七提取物通过上调VEGF、KDR、FLT-1和HIF-1表达促进血管生成，是其治疗UC的主要作用机制之一。     

Curcumin-attenuated trinitrobenzene sulphonic acid induces chronic colitis by inhibiting expression of cyclooxygenase-2

AIM: To explore the possible mechanisms of curcumin in rat colitis induced by tri nitrobenzene suifonic (TNBS) acid. METHODS: Rats with TNBS acid-induced colitis were treated with curcumin (30 mg/kg or 60 mg/kg per day ip). Changes of body weight and histoiogicai scores as well as survival rate were evaluated. Leukocyte infiltration was detected by myeloperoxidase (MPO) activity assay. The expression of cyclooxygenase-2 (COX-2) was detected by RT-PCR and Western blot. Inflammation cytokines were determined by RT-PCR. Local concentration of prostaglandin E2 (PGE2) in colon mucosa was determined by ELISA. RESULTS: Curcumin improved survival rate and histoiogicai image, decreased the macroscopic scores and MPO activity. Also curcumin reduced the expression of COX-2 and inflammation cytokines. In addition, treatment with curcumin increased the PGE2 level. CONCLUSION: Curcumin has therapeutic effects on TNBS acid-induced colitis, the mechanisms seem to be related to COX-2 inhibition and PGE2 improvement.     

白细胞介素-23/-17轴在小鼠急性实验性结肠炎中的表达及作用

目的 研究白细胞介素(IL)-23/IL-17轴在小鼠实验性结肠炎结肠组织中的表达和作用.方法 将64只小鼠分为对照组24只、模型组24只、抗体组8只、正常血清组8只.除对照组外,其余各组建立小鼠急性实验性结肠炎模型.对照组和模型组小鼠分别于造模后24 h、48 h、7 d处死.抗体组和正常血清组小鼠分别于造模前2 h腹腔内注射多克隆大鼠抗小鼠IL-17中和抗体和正常大鼠血清,于造模48 h后处死.检测各组小鼠组织学损伤评分、肠组织髓过氧化物酶(MPO)活性;酶联免疫吸附试验检测结肠组织IL-23p19、IL-17含量;免疫组化染色检测核因子(NF)-κB p65在结肠组织中的表达;实时荧光定量(RT)PCR检测IL-23p19、IL-17、IL-12p35的mRNA表达水平.结果 模型组24 h、48 h、7 d时IL-23p19蛋白表达水平和mRNA表达水平[分别为(15.53±3.32)、(31.16±4.98)、(14.03±3.56)ng/mg蛋白和4.09±0.34、3.39±0.46、6.54±1.82]、IL-17的蛋白表达水平和mRNA表达水平[分别为(0.35±0.06)、(0.38±0.08)、(0.26±0.05)ng/mg蛋白和4.21±2.61、2.65±0.91、5.63±1.43]均显著高于正常对照组(P值均＜0.05),48 h时达高峰.IL-23与IL-17蛋白表达水平和mRNA表达水平呈正相关(r值分别为0.745和0.793,P＜0.05).抗体组IL-23p19和IL-12p35高水平表达,但NF-κB p65阳性细胞率、组织学评分及MPO活性[分别为1.86%±0.36%、0.63±0.52、(0.40±0.03)U/g]明显低于48 h模型组[分别为4.35%±0.37%、5.13±0.64、(2.29±0.40)U/g],说明中和IL-17后能明显减轻结肠炎症,抑制NF-κB活性.结论 IL-23/IL-17轴在急性实验性结肠炎早期阶段起关键作用.IL-17有望成为炎症性肠病治疗的新靶标.     

T细胞亚群在TNBS诱发大鼠实验性结肠炎发病及治疗后的变化

目的探讨T细胞亚群在TNBS诱发大鼠实验性结肠炎模型的外周血、脾、结肠中变化特点及经柳氮磺胺吡啶（SASP）和氢化泼尼松（PSL）治疗后这些细胞的变化。方法分为对照组、建模后1周及3周组、SASP及PSL治疗组，用流式细胞仪检测各组外周血、脾和结肠黏膜上皮细胞内CD4^＋，CD8^＋T细胞比例的变化；肠道积分评定组织学变化。结果CD4^＋T细胞：建模第1周外周血、脾明显低于对照组；建模第3周脾CD4^＋T细胞升至对照组水平，SASP治疗组外周血高于模型3周组，PSL治疗组外周血、脾、结肠均明显低于对照组和模型3周组。CD8^＋T细胞：建模第1周外周血、结肠低于对照组，建模第3周外周血中仍低，结肠升至对照组水平；SASP治疗组外周血高于模型3周组。PSL治疗组外周血、脾、结肠均明显低于对照组，脾、结肠低于模型3周组。结论大鼠实验性结肠炎发病过程中，机体免疫功能下调，以局部免疫异常为主，与整体的免疫异常可能关系较小。PSL导致机体免疫功能下调。SASP对全身免疫状态影响较小，其作用更集中于肠道。     

Melatonin reduces bacterial translocation and apoptosis in trinitrobenzene sulphonic acid-induced colitis of rats

AIM： To investigate the effects of exogenous melatonin on bacterial translocation and apoptosis in a rat ulcerati-ve colitis model. METHODS： Rats were randomly assigned to three groups： groupⅠ： control, group Ⅱ： experimental colitis, group Ⅲ： colitis plus melatonin treatment. On d 11 after colitis, plasma tumor necrosis factor-α, portal blood endotoxin levels, colon tissue myeloperoxidase and caspase-3 activity were measured. Bacterial translocation was quantified by blood, lymph node, liver and spleen culture. RESULTS： We observed a significantly reduced inciden-ce of bacterial translocation to the liver, spleen, mesen-teric lymph nodes, portal and systemic blood in animals treated with melatonin. Treatment with melatonin signifi-cantly decreased the caspase-3 activity in colonic tissues compared to that in trinitrobenzene sulphonic acid-trea-ted rats （16.11 ± 2.46 vs 32.97 ± 3.91, P 〈 0.01）. CONCLUSION： Melatonin has a protective effect on ba-cterial translocation and apoptosis.     

Ameliorative effects of bombesin and neurotensin on trinitrobenzene sulphonic acid-induced colitis, oxidative damage and apoptosis in rats

AIM： TO investigate the effects of bombesin （BBS） and neurotensin （NTS） on apoptosis and colitis in an ulcerative colitis model.
METHODS： In this study, a total of 50 rats were divided equally into 5 groups. In the control group, no colitis induction or drug administration was performed. Colitis was induced in all other groups. Following the induction of colitis, BBS, NTS or both were applied to three groups of rats. The remaining group （colitis group） received no treatment. On the 11th d after induction of colitis and drug treatment, blood samples were collected for TNF-α and IL-6 level studies. Malondialdehyde （MDA）, carbonyl, myeloperoxidase （MPO） and caspase-3 activities, as well as histopathological findings, evaluated in colonic tissues.
RESULTS： According to the macroscopic and microscopic findings, the study groups treated with BBS, NTS and BBS ＋ NTS showed significantly lower damage and inflammation compared with the colitis group （macroscopic score, 2.1 ± 0.87, 3.7 ± 0.94 and 2.1 ± 0.87 vs 7.3 ± 0.94; microscopic score, 2.0 ± 0.66, 3.3 ± 0.82 and 1.8 ± 0.63 vs 5.2 ± 0.78, P 〈 0.01）. TNF-α and IL-6 levels were increased significantly in all groups
compared with the control group. These increases were significantly smaller in the BBS, NTS and BBS ＋ NTS groups compared with the colitis group （TNF-α levels, 169.69 ± 53.56, 245.86 ± 64.85 and 175.54 ± 42.19 vs 556.44 ± 49.82; IL-6 levels, 443.30 ± 53.99, 612.80 ± 70.39 and 396.80 ± 78.43 vs 1505.90 ± 222.23, P 〈 0.05）. The colonic MPO and MDA levels were significantly lower in control, BBS, NTS and BBS ＋ NTS groups than in the colitis group （MPO levels, 24.36 ± 8.10, 40.51 ± 8.67 and 25.83 ± 6.43 vs 161.47 ± 38.24; MDA levels, 4.70 ± 1.41, 6.55 ± 1.12 and 4.51 ± 0.54 vs 15.60 ± 1.88, P 〈 0.05）. Carbonyl content and caspase-3 levels were higher in the colitis and NTS groups than in control, BBS and BBS ＋ NTS groups （carbonyl levels, 553.99 ± 59.58 and 336.26 ± 35.72 vs 209.7     

三硝基苯磺酸/乙醇与免疫复合物联合诱导的结肠炎动物模型

目的 建立三硝基苯磺酸/乙醇与免疫复合物联合诱导的结肠炎动物模型,探讨炎症性肠病的发病机制.方法 SD大鼠168只,雌雄各半,分为:①正常对照组42只,②三硝基苯磺酸/乙醇模型组42只,③免疫复合物组42只,④三硝基苯磺酸/乙醇与免疫复合物联合诱导的结肠炎动物模型组42只(以上每组雌雄各半).分别于造模后1、2、3、4、6,9、12周末将大鼠处死(每组每周处死6只).观察各组结肠大体形态和组织学改变,并检测肠黏膜髓过氧化物酶(MPO)活性,大便潜血实验.结果 三硝基苯磺酸/乙醇与免疫复合物联合诱导的结肠炎动物模型组大鼠大体形态损伤评分均数为5.8571±0.8431;组织学损伤评分均数为4.8571±0.3542;MPO均数为3.8588±0.8625;OBT积分均数为4.1429±0.6010皆明显高于其余各组,具有统计学意义.结论 上述联合诱导的结肠炎动物模型是一种较理想的IBD动物模型,可作为研究IBD发病机制及评估药物疗效的有益工具.     

Role of inducible nitric oxide synthase in trinitrobenzene sulphonic acid induced colitis in mice

BACKGROUND: Studies using inhibitors of nitric oxide synthase (NOS) to date are inconclusive regarding the role of inducible NOS (iNOS) in intestinal inflammation. AIMS: (1) To examine the role of iNOS in the development of chronic intestinal inflammation; (2) to identify the cellular source(s) of iNOS. METHODS: Colitis was induced by an intrarectal instillation of trinitrobenzene sulphonic acid (TNBS, 60 mg/ml, 30% ethanol), in wild type (control) or iNOS deficient mice. Mice were studied over 14 days; the colons were scored for injury and granulocyte infiltration was quantified. Blood to lumen leakage of (51)Cr-EDTA was measured as a quantitative index of mucosal damage. RESULTS: At 24 and 72 hours, iNOS deficient mice had significantly increased macroscopic inflammation compared with wild type mice. Granulocyte infiltration increased significantly at 24 hours and remained elevated in iNOS deficient mice at 72 hours, but significantly decreased in controls. However, by seven days post-TNBS macroscopic damage, microscopic histology, granulocyte infiltration, and mucosal permeability did not differ between wild type and iNOS deficient mice. A four- to fivefold increase in iNOS mRNA was observed in wild type mice at 72 hours and seven days post-TNBS and was absent in iNOS deficient mice. Immunohistochemistry techniques showed that iNOS expression was predominantly localised in neutrophils, with some staining also in macrophages. CONCLUSIONS: These results suggest that leucocyte derived iNOS ameliorates the early phase, but does not impact on the chronic phase of TNBS induced colitis despite the presence of iNOS.