葡聚糖硫酸钠诱导动物溃疡性结肠炎及其机制

溃疡性结肠炎(UC)系由机体局部免疫器官对肠道多种抗原的高敏状态而引起的非特异性炎症,属自发性炎症性肠病(IBD)。鉴于其致病机制尚待探讨且国内报道少见,本文采用葡聚糖硫酸钠(DSS)诱导动物UC,分析比较实验动物病理、生化及组化指标,探讨了模型建立的方法及发病机制。 1 材料与方法 1.1 材料 (1)动物:雌性BALB/c小鼠,体重20 g。Harty's豚鼠,雌雄各半,体重280～310 g。均购自中山医科大学动物实验中心,具Ⅰ级动物合格证。恒温常规饮食,豚     

黄芩素对葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠的改善作用

目的:探究黄芩素(baicalein)对葡聚糖硫酸钠(dextran sulfate sodium salt, DSS)诱导的溃疡性结肠炎(ulcerative colitis, UC)小鼠的疗效及可能的作用机制。方法:将42只雄性BALB/c小鼠随机分为6组(n=7),分别为空白组,模型组,黄芩素低、中、高剂量组和美沙拉嗪阳性药物对照组。采用4%的DSS创建小鼠UC模型,造模成功后,空白组和模型组自由饮用蒸馏水,黄芩素低、中、高剂量组分别灌胃(ig)10,20,40 mg·kg^-1黄芩素混悬液,阳性药物组给予美沙拉嗪600 mg·kg^-1,ig,连续治疗14 d。计算并评价疾病活动指数(DAI);采用苏木精-伊红染色法(HE)观察小鼠结肠组织病理学变化;采用酶联免疫吸附测定(ELISA)试剂盒定量检测血清中脂多糖(LPS)、分泌型免疫球蛋白A(sIg A)水平;采用蛋白免疫印迹法(Western blot)检测各组小鼠结肠组织中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和核转录因子-κB(NF-κB)的表达。结果:黄芩素可降...     

复方肠泰对葡聚糖硫酸钠诱导的溃疡性结肠炎治疗作用的研究

目的 探讨复方肠泰对葡聚糖硫酸钠(DSS)诱导的小鼠慢性溃疡性结肠炎(UC)的治疗作用。方法 随机将C57BL/6小鼠分为正常对照组、DSS模型组、复方肠泰给药组。采用2% DSS溶液对小鼠进行造模,小鼠连续5 d自由饮用DSS水,接着连续5 d自由饮水,重复3个循环。复方肠泰组每天按照18 g/kg ig给药,其他组ig给予相同体积的蒸馏水。每天记录小鼠体质量、疾病活动指数(DAI)评分结果,采用HE染色观察结直肠组织病理学变化,给予组织病理学评分;ELISA检测TNF-α、IL-10表达水平,免疫组化检测COX-2蛋白表达情况。结果 与DSS模型组比较,复方肠泰组小鼠疾病状态得到明显改善,DAI和组织病理学评分显著降低,炎症因子IL-10表达升高,TNF-α、COX-2蛋白的表达降低。结论 复方肠泰对小鼠慢性UC具有显著地治疗作用,可能通过促进抗炎症因子IL-10的表达,同时抑制促炎症因子TNF-α、COX-2蛋白的表达发挥作用。     

结肠炎奇效颗粒对葡聚糖硫酸钠所致溃疡性结肠炎大鼠的影响

摘 要 目的：观察结肠炎奇效颗粒对实验性溃疡性结肠炎模型大鼠的治疗作用。方法: 180～220g Wistar大鼠,采用饮用4%葡聚糖硫酸钠溶液方法制备大鼠实验性溃疡性结肠炎（UC）模型,按2.70,1.35,0.68 g·kg^-1·d^-1剂量灌胃给予结肠炎奇效颗粒7 d后,分别检测血清丙二醛(MDA)、超氧化物歧化酶(SOD)、一氧化氮(NO)、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）及结肠的髓过氧化物酶（MPO）、结肠黏膜细胞间黏附分子（ICAM-1）的蛋白表达和核因子（NF-kBp65）的蛋白表达。 结果: 与模型组相比,结肠炎奇效颗粒高、中剂量组明显提高结肠炎大鼠血中SOD活性（P<0.05或P<0.01）,显著降低MDA、TNF-α、IL-6的含量（P<0.05或P<0.01）;明显减低溃疡性结肠炎大鼠结肠黏膜中MPO活性（P<0.05或P<0.01）;显著降低结肠炎大鼠结肠炎症反应中核因子蛋白（NF-kBp65）和细胞黏附因子（ICAM-1）蛋白含量（P<0.01）。结论:结肠炎奇效颗粒对葡聚糖硫酸钠溶液所致溃疡性结肠炎模型大鼠治疗作用明显。     

葡聚糖硫酸钠自由饮用与灌胃诱导小鼠溃疡性结肠炎模型的比较

目的比较不同浓度葡聚糖硫酸钠(dextran sulfate sodium,DSS)在自由饮用和灌胃两种方式下诱导小鼠急性溃疡性结肠炎(ulcerative colitis,UC)成模情况和模型相关指标变异系数,优选UC造模方法。方法小鼠随机分为正常组、3%DSS溶液自由饮用组、4%DSS溶液自由饮用组、30%DSS溶液灌胃组、40%DSS溶液灌胃组。灌胃组晚于自由饮用组1 d开始给予DSS,30%DSS灌胃组和40%DSS溶液灌胃组的灌胃剂量分别与3%DSS饮用组、4%DSS饮用组小鼠前1 d摄入DSS的平均剂量相等。观察记录小鼠体质量、食量、疾病活动指数(DAI)、主要脏器指数、结肠长度及组织病理学评分,比较不同造模方法上述各指标变异系数。结果4种造模方式均能成功诱导UC。其中,3%DSS自由饮用组在动物存活率、造模成本和操作方便程度方面优于其他造模方法,各指标变异系数未明显高于其他方法。结论3%DSS溶液自由饮用法是本实验中小鼠急性UC较合适的造模方法。     

葡聚糖硫酸钠自由饮用与灌胃诱导小鼠溃疡性结肠炎模型的对比研究

摘 要 目的：比较葡聚糖硫酸钠（DSS）自由饮用与灌胃两种给药方式诱导小鼠溃疡性结肠炎（UC）疾病相关指标的差异,为优化UC小鼠造模方法提供实验参考。方法: 将C57BL/6小鼠随机分为正常对照组、自由饮用组和灌胃组3组,每组10只,分别自由饮水、自由饮用3% DSS溶液及4 g·kg^-1·d^-1灌胃DSS,连续7 d,建立UC模型。以疾病活动指数（DAI）、小鼠结肠的组织学损伤评分及髓过氧化物酶（MPO）活性为评价指标,比较两种给药方式的各指标均数及变异系数的差异。结果: 自由饮用组造模过程中有2只小鼠死亡,造模结束后存活小鼠的DAI值为（8.8±1.6）分,灌胃组无小鼠死亡,其DAI值为（9.0±0.8）分,两组间差异无统计学意义（P＞0.01）,但自由饮用组的变异系数高于灌胃组（18.7 vs. 8.6）。自由饮用组与灌胃组结肠组织学损伤评分分别为（24.8±4.2）,（27.0±2.8）分,均呈典型性炎症改变,两组间差异无统计学意义（P＞0.05）,但自由饮用组的变异系数高于灌胃组（16.9 vs. 10.4）。正常对照组、自由饮用组与灌胃组结肠组织MPO分别为（0.41±0.03）,（2.32±0.34）,（2.05±0.18）U·g^-1,与正常对照组相比,自由饮用组与灌胃组的MPO差异有统计学意义（P＜0.01）,自由饮用组与灌胃组两组间MPO差异无统计学意义（P＞0.05）,但自由饮用组的变异系数高于灌胃组（14.7 vs. 8.8）。结论:自由饮用组与灌胃组均可成功诱导小鼠UC模型。与自由饮用组相比,灌胃组各项指标数据的变异系数小,离散度低,采用灌胃法制备溃疡性结肠炎小鼠模型,动物表现更均一。     

葡聚糖硫酸钠诱导不同性别溃疡性结肠炎小鼠模型的比较

目的:探讨性别差异对小鼠溃疡性结肠炎(UC)模型成模的影响,为实验造模提供参考依据.方法:将24只C57BL/6J小鼠根据性别随机分为4组,雌性对照组、雄性对照组、雌性DSS组和雄性DSS组,每组6只,对照组正常饮食,雌性或雄性DSS组给予正常饮食+3％葡聚糖硫酸钠(DSS)溶液自由饮用7?d进行造模.每日记录各组小鼠...     

吲哚丙酸对葡聚糖硫酸钠诱导急性溃疡性结肠炎小鼠的影响

目的 研究吲哚丙酸(IPA)对葡聚糖硫酸钠(DSS)诱导急性溃疡性结肠炎(UC)小鼠的影响。方法将SPF级6～8周雄性C57BL/6小鼠随机分为对照组、DSS模型组(饮用3%DSS 7 d)及低、中、高剂量IPA组(在DSS给药前2 d行IPA灌胃),每组12只。每日观察小鼠的一般情况,根据体质量下降、大便性状和便血状况评定疾病活动指数(DAI); DSS给药第7日处死小鼠,测量结肠长度及评定病理学评分,ELISA检测各组小鼠血清髓过氧化物酶(MPO)及结肠组织MPO、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)水平。Western blot法检测各组小鼠结肠组织中核因子-κB p65(NF-κB p65)、磷酸化NF-κB p65(p-NF-κB p65)蛋白表达。结果 与对照组比较,DSS模型组DAI、结肠组织病理学评分、血清MPO及结肠组织TNF-α、IL-1β、IL-6、MPO和p-NF-κB p65/NF-κB p65比值显著升高,结肠长度明显缩短(P <0.05,P <0.01);与DSS模型组比较,中、高剂量IPA...     

盐酸小檗碱对葡聚糖硫酸钠所致小鼠结肠炎的作用研究

目的研究盐酸小檗碱对小鼠实验性溃疡性结肠炎的保护作用。方法实验设正常、模型、柳氮磺胺吡啶(SASP,520 mg·kg-1)和盐酸小檗碱(berberine chloride,BER)低、中、高(15 mg·kg-1、45 mg·kg-1、150 mg·kg-1)剂量组。六组小鼠自由饮用4%葡聚糖硫酸钠(dextran sulphate sodium, DSS)水溶液或蒸馏水,同时分别灌胃给予干预药物或溶剂(0·2 mL / 10 gWt,1次/ d×7 d)。以疾病活动指数(Disease activity index, DAI)评价小鼠临床症状。生化方法测定结肠组织MDA含量,SOD、MPO活性;HE染色切片,评估结肠组织病理改变;免疫组化检测ICAM-1、NF-κB p65蛋白表达水平。结果模型组DAI显著增高,结肠黏膜损伤严重;MDA含量、MPO活性及ICAM-1和NF-κB p65表达明显升高,SOD活性下降。SASP 520 mg·kg-1能明显逆转上述情况改变;BER使DAI减小,结肠黏膜损伤减轻, MDA含量、MPO活性降低,阻遏ICAM-1和NF-κB p65蛋白表达,升高SOD活性,且呈现一定剂量依赖性。结论BER可能通过抗氧自由基作用,抑制ICAM-1表达及NF-κB激活,缓解DSS所致的小鼠结肠炎症。     

覆盆子改善溃疡性结肠炎小鼠模型的作用研究

研究覆盆子对溃疡性结肠炎小鼠的治疗作用及其相关机制。用葡聚糖硫酸钠(Dextran sulphate sodium, DSS)诱导建立溃疡性结肠炎小鼠模型,将溃疡性结肠炎小鼠随机分为正常对照(NC)组、结肠炎(DSS)组、覆盆子(DSS+BB)治疗组、覆盆子对照(BB)组。其中,DSS组、DSS+BB组自由饮用2.5%DSS溶液1 w,NC组、BB组饮用正常灭菌水。期间,用中药材覆盆子(200 mg/kg)灌胃DSS+BB组及BB组,每日1次,连续1 w。实验期间,每日观察小鼠进食、活动、毛发等一般情况,检测体重,观察粪便形态。实验第8 d,异氟烷麻醉小鼠,取血后处死小鼠,取血清进行炎症因子白介素6(IL-6)检测,结肠HE染色行形态学观察。覆盆子对正常和实验动物体重无明显影响,DSS诱导的炎症状态引发明显的体重下降(P<0.05);覆盆子能明显改善DSS诱导的结肠炎性改变;覆盆子对DSS诱导的循环和局部炎症因子IL-6有明显下调作用(P<0.01)。覆盆子可明显改善溃疡性结肠炎的表型并具有明显的抗炎作用。     

改良愈疡方对溃疡性结肠炎模型小鼠抗炎作用机制的研究

目的探讨改良愈疡方对右旋葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎模型小鼠抗炎作用机制。方法40只雄性C57BL／6小鼠除正常组外,采用口服DSS造模后,按完全随机法分为模型组、治疗组、对照组,每组10只;正常组和模型组均用0．9％氯化钠溶液灌肠,治疗组动物给与中药灌肠,对照组给与柳氮磺胺吡啶灌肠共14d。进行结肠肿瘤坏死因子（TNF）-α水平测定及通过免疫组化检测TNF-α在结肠组织中的表达。结果对照组、治疗组治疗前与模型组基本相同,治疗后治疗组的大便较成形、血便及体质量减轻较不明显,一般情况优于模型组及对照组（P＜0．05）。SS小鼠结肠炎结肠组织中TNF-α较正常组显著增高（P＜0．05）,用中药治疗组则显著降低,且优于对照组（P＜0．05）。同时,在模型组小鼠结肠组织中,TNF—α表达主要见于黏膜和黏膜下层,主要集中在靠近糜烂或溃疡的周边,小血管扩张的周围,也相对增多,治疗组TNF-α阳性细胞明显减少,与模型组及对照组比较均有显著差异（P〈0．05）。结论改良愈疡方对溃疡性结肠炎模型小鼠具有良好的治疗作用,其作用机制是通过降低TNF-α含量。抑制炎症有关。     

Anti-inflammatory effects of Vicenin-2 on dextran sulfate sodium-induced colitis in mice

The objective of the present work was to evaluate the anti-inflammatory effects of Vicenin-2 on dextran sulfate sodium (DSS)-induced colitis model. Colitis was induced in C57BL/6J mice by administration of 2% DSS in drinking water for 7 days. In addition to DSS, Vicenin-2 (50 mg kg⁻¹/day⁻¹) was administrated orally to the test group. The ulceration extent and severity were assessed macroscopically, histopathologically, and by disease activity index. The Vicenin-2 treated group showed significant differences in physiological parameters including bodyweight, colon weight, and colon length, compared to DSS-induced colitis group. In addition, Vicenin-2 treatment effectively reduced stool consistency and bleeding scores. Myeloperoxidase (MPO) activity, expressions of pro-inflammatory cytokines, and specific key inflammatory markers (iNOS and COX-2) significantly increased in DSS-induced colitis colon tissues. However, administration of Vicenin-2 effectively reduced the MPO activity, attenuated the expression of pro-inflammatory cytokines and key inflammatory markers, in DSS-induced colitis mice. These results were comparable with sulfasalazine, an anti-inflammatory drug used routinely for ulcerative colitis (UC). These findings suggest that Vicenin-2 effectively suppresses DSS-induced colitis by attenuating expressions of key inflammatory mediators and found to be an attractive therapeutic drug for treating UC.     

Prophylactic role of curcumin in dextran sulfate sodium (DSS)-induced ulcerative colitis murine model

We have addressed in this study the possible protective role of the main principle of turmeric pigment; curcumin on a murine model of ulcerative colitis (UC). Colitis was induced by administration of dextran sulfate sodium (DSS) (3% W/V) in drinking water to male Swiss albino rats for 5 consecutive days. DSS challenge induced UC model that was well characterized morphologically and biochemically. DSS produced shrinkage of colon length and increased the relative colon weight/length ratio accompanied by mucosal edema and bloody stool. Histologically, DSS produced submucosal erosions, ulceration, inflammatory cell infiltration and crypt abscess as well as epithelioglandular hyperplasia. The model was confirmed biochemically, and the test battery entailed elevated serum tumor necrosis factor (TNF-α) and colonic activity of myleoperoxidase (MPO). Colonic glutathione-S-transferase (GST) activity and its substrate concentration; GSH, were notably reduced, while lipid peroxidation, expressed as malondialdehyde (MDA) level, and total nitric oxide (NO) were significantly increased. Prior administration of curcumin (100 mg/kg, IP) for 7 consecutive days ahead of DSS challenge mitigated the injurious effects of DSS and ameliorated all the altered biochemical parameters. These results suggest that curcumin could possibly have a protective role in ulcerative colitis probably via regulation of oxidant/anti-oxidant balance and modulation of the release of some inflammatory endocoids, namely TNF-α and NO.     

A new flavonoid derivative, dosmalfate, attenuates the development of dextran sulphate sodium-induced colitis in mice

In this study, we have evaluated the efficacy of dosmalfate, a new flavonoid derivative compound, for the prevention and treatment of experimental colitis. To induce colitis, BALB/c mice received 5% dextran sulphate sodium (DSS) in their drinking water continuously for 7 days. Colitis was quantified by a clinical damage score, colon length, weight loss, stool consistency and rectal bleeding. Inflammatory response was assessed by neutrophil infiltration, determined by histology and myeloperoxidase (MPO) activity. Interleukin (IL)-1 beta, prostaglandins (PG)E(2) and (PG)D(2) concentrations in colonic tissue, histological and histochemical analysis of the lesions were also measured. Dosmalfate (400-800 mg/kg body weight, p.o.) ameliorated severe colitis reduced the degree of inflammation through reduction of neutrophil infiltration and IL-1 beta levels. (PG)E(2) and (PG)D(2) synthesis were significantly reduced in colitis control group and treatment with dosmalfate abolished the decrease in PG synthesis in colon mucosa. We conclude that dosmalfate is protective in acute DSS-induced colitis. The beneficial effects seem to be related to a decrease of neutrophil infiltration, absence of up-regulation of IL-1 beta and increase of PG production in colon mucosa.     

The natural flavonoid galangin ameliorates dextran sulphate sodium–induced ulcerative colitis in mice: Effect on Toll‐like receptor 4, inflammation and oxidative stress

This study was carried out to investigate the potential therapeutic effect of galangin, a promising active principle of honeybee propolis, in dextran sulphate sodium (DSS)–induced colitis in mice. We explored the possible underlying mechanisms for galangin action and the therapeutic benefit of adding galangin to the standard therapy sulphasalazine. A galangin dose of 40 mg/kg was selected based on a preliminary dose‐selection study for investigation in a 4‐week cyclical model of DSS‐induced colitis. Mice received 3% DSS in their drinking water during the first and third weeks and were administered the treatments (40 mg/kg galangin, 100 mg/kg sulphasalazine and a combination of 20 mg/kg galangin and 50 mg/kg sulphasalazine) daily starting from the second week. Galangin significantly ameliorated DSS‐induced histopathological alterations and tissue injury, down‐regulated Toll‐like receptor 4 expression, suppressed NF‐κB p65 activation, lowered inflammatory cytokine levels and demonstrated antioxidant effects. The combination of galangin and sulphasalazine at half doses yielded comparable results to either drug alone at full dose. This study highlights galangin as a promising therapy for colitis management.     

灰绿黄堇总生物碱对实验性溃疡性结肠炎的作用研究

目的研究灰绿黄堇总生物碱（ACAM）对小鼠结肠炎的作用及其机制。方法实验设正常、模型、柳氮磺胺吡啶组（SASP,520mg/kg）和ACAM组（100、200、400mg/kg）。正常组小鼠饮用蒸馏水,其余组自由饮用4%葡聚糖硫酸钠（DSS）水溶液,同时分别灌胃给予溶剂或干预药物（0.2ml/10g,1次/d×7d）。记录小鼠疾病活动指数（DAI）;测定结肠组织MDA含量,SOD、MPO活性及ICAM-1、NF-κBp65表达水平。结果模型组DAI显著增高,结肠黏膜损伤严重;MDA舍量、MPO活性及ICAM-1和NF-κBp65表达明显升高。SOD活性下降（P〈0.01）。SASP520mg/kg能明显逆转上述改变;ACAM100mg具有相似的作用。结论ACAM可能通过抗氧自由基作用,抑制炎性细胞活化、迁移及NF-κB激活,缓解小鼠结肠炎性反应。     

烟酰胺单核苷酸对小鼠溃疡性结肠炎的作用

目的 探究烟酰胺单核苷酸(NMN)对葡聚糖硫酸钠盐(DSS)诱导的溃疡性结肠炎小鼠模型的作用.方法 采用DSS诱导溃疡性结肠炎小鼠模型评价NMN的作用.应用DSS对C57小鼠进行造模,给予NMN后观察模型小鼠的生存时间、体重、疾病评分(DAI)、结肠组织长度以及结肠组织切片病理学变化.结果 NMN对模型小鼠的生存时间、...