SP和CGRP在炎症性肠病大鼠结肠中的表达规律

目的通过研究P物质（SP）和降钙素基因相关肽（CGRP）在炎症性肠病（IBD）大鼠结肠中的表达规律来探讨二者在IBD发生发展过程中发挥的作用。方法采用三硝基苯磺酸（TNBS）化学诱导建立IBD大鼠动物模型,应用real-time RT-PCR方法来检测SP和CGRP mRNA在模型不同时期结肠中的表达变化。结果随着炎症性肠病病理变化的加剧（第3、7天）,sP和CGRP在结肠组织中的表达水平显著升高;随着疾病的恢复（第21、28天）,其表达水平逐渐降低并趋向于正常水平。从总体来看,在IBD大鼠结肠组织中,SP和CGRP mRNA水平的变化与IBD的发生、发展呈正相关。结论神经递质SP和CGRP在IBD炎症和修复中起重要的神经免疫信号传导作用。     

三硝基苯磺酸诱导的C57/BL6小鼠结肠炎模型的建立方法

2,4,6-三硝基苯磺酸(TNBS)诱导的小鼠结肠炎模型是研究人类炎性肠病(inflammatorybowl disease,IBD)的主要手段之一,但在实际应用中,常用的C57/BL品系小鼠却对TNBS有较高耐受性,不易建模.本文主要介绍一种可以有效诱导C57BL6小鼠TNBS结肠炎的方法,并对疾病评价指标进行了具体...     

炎症性肠病小鼠模型的建立及其影响因素探讨

目的:通过单次灌肠和皮肤致敏联合灌肠,构建2,4,6-三硝基苯磺酸(TNBS)诱导的炎症性肠病小鼠模型,探讨最佳造模方法,并分析影响模型构建的因素。方法:55只SPF雄性BALB/c小鼠随机分为7组,包括对照组、不同剂量TNBS(100、150、175、200、225 mg/kg)单次灌肠组及皮肤致敏联合灌肠组。于造模后5 d处死各组小鼠,观察结肠大体形态并评分;取病变处进行石蜡包埋切片,HE染色,并进行病理组织学评分。结果:100、150 mg/kg TNBS单次灌肠组动物未见明显的溃疡形成;其余剂量组动物均有不同程度的溃疡形成,成模率与剂量成正比,其中225 mg/kg剂量组动物成模率为100%,但病变较重、病变不均一且偶有小鼠眼睛失明的副作用出现。皮肤致敏联合灌肠组动物均有溃疡形成,成模率100%,病变适中且未发现小鼠眼睛失明的副作用。结论:175-225 mg/kg TNBS单次灌肠及皮肤致敏联合TNBS灌肠均可制备小鼠炎症性肠病模型,但皮肤致敏联合TNBS灌肠制备的炎症性肠病模型成模率高,病变适中,模型稳定,适合用作科学研究模型。     

小鼠动物实验方法系列专题(三) 三硝基苯磺酸诱导的C57/BL6小鼠结肠炎模型的建立方法

2,4,6-三硝基苯磺酸(TNBS)诱导的小鼠结肠炎模型是研究人类炎性肠病(inflammatory bowl disease,IBD)的主要手段之一,但在实际应用中,常用的C57/BL品系小鼠却对TNBS有较高耐受性,不易建模。本文主要介绍一种可以有效诱导C57/BL6小鼠TNBS结肠炎的方法,并对疾病评价指标进行了具体的描述。对基因工程小鼠IBD模型的研究具有重要意义。     

CT仿真结肠镜与腹部增强CT 在炎症性肠病诊断中的对比分析

目的:对比CT仿真结肠镜(CTC)与腹部增强CT在炎症性肠病诊断中的价值,为临床诊疗提供依据。方法:选取2015年1月到2016年1月我院收治的炎症性肠病患者60例为研究组,另选取非炎症性肠病患者60例为对照组,两组均30例患者接受CTC检查,30例患者接受腹部增强CT检查,对影像图片进行盲法阅读,将炎症性肠病的可信度分为5个等级进行评价,对6项影像学特征进行Logistic回归分析。结果:腹部增强CT对炎症性肠病诊断的准确率、敏感性、特异性分别为68.33%,59.09%,71.39%显著低于CTC的93.33%,95.01%、89.63%,比较差异具有统计学意义(P0.05);Logistic回归分析显示:结肠粘膜呈颗粒状、不光滑;肠壁强化并增厚;结肠袋消失对炎症性肠病具有预测作用;肠壁强化及增厚对炎症性肠病诊断敏感性较高,结肠粘膜呈颗粒状、不光滑联合结肠袋消失诊断炎症性肠病的特异性较高。结论:CTC对炎症性肠病具有较高的检出率,肠壁强化及增厚对炎症性肠病诊断的敏感性较高,结肠粘膜呈颗粒状、不光滑联合结肠袋消失诊断炎症性肠病特异性较高。     

人脐带间充质干细胞对炎症性肠病小鼠模型的治疗作用

目的:探讨2种方式移植人脐带间充质干细胞(UCMSC)对TNBS诱导的炎症性肠病小鼠模型的治疗作用。方法:分离培养人UCMSC,并通过一次性腹腔注射TNBS制备炎症性肠病动物模型,采用尾静脉注射和腹腔注射2种方式移植人UCMSC进行治疗,4周后处死,观察动物的死亡率、体重改变及结肠炎症变化,并进行大体和病理评分。结果:与对照组相比,尾静脉和腹腔注射移植UCMSC组动物死亡率下降,体重恢复较早,结肠病变大体评分和病理评分均显著改善;2种方式移植组之间的评分无显著差异。结论:尾静脉和腹腔移植人UCMSC对TNBS诱导的炎症性肠病小鼠模型均有治疗作用。     

布拉酵母对炎症性肠病的治疗作用

炎症性肠病包括溃疡性结肠炎和克罗恩病, 其病因与发病机制尚未完全明确。大量研究表明, 肠道微生物在炎症性肠病的发生、发展中发挥重要作用。布拉酵母是一种有益于人体健康的肠道微生物, 研究发现能有效改善炎症性肠病症状, 可能与其抑制肠道致病菌、增强肠屏障功能、调节肠道黏膜免疫反应等有关。     

氨基酸对炎症性肠病的调控作用北大核心CSCD

炎症性肠病是胃肠道的一种慢性复发性炎症,包括克罗恩病和溃疡性结肠炎,患者众多,而且目前很难实现彻底治愈。由于患者消化功能受损,食物不容易吸收,很容易出现营养不良的情况,临床经常使用营养治疗来克服营养不足、改变炎症状态。氨基酸作为辅助营养治疗,可能有助于维持炎症性肠病患者的肠道完整性,减少炎症、氧化应激和肠道细胞死亡,对炎症性肠病的治疗起到积极的作用。最近在动物方面的研究已经证明氨基酸在炎症性肠病治疗中存在着巨大的潜力,氨基酸的供应和代谢可能是一种有前景的辅助治疗方法。本文就谷氨酰胺、精氨酸、甘氨酸等特定氨基酸的免疫调节作用进行综述,以期提供一种新的治疗炎症性肠病的思路。     

核酸药物及其给药系统用于炎症性肠病治疗的研究进展

炎症性肠病是一种常见的免疫功能紊乱所致慢性顽固性胃肠道炎性疾病,现有的治疗手段难以根治。随着炎症性肠病分子机制研究的不断深入,在基因水平上应用核酸药物及其给药系统,对炎症性肠病发挥的独特治疗作用,已受到越来越多的关注, 并取得一定进展。本文简介炎症性肠病的发病机制,综述近年来核酸药物及其给药系统用于炎症性肠病治疗的研究进展。     

益生菌对炎症性肠病幼鼠 Th17细胞的调节作用

目的观察鼠李糖乳杆菌(LGG)对炎症性肠病(IBD)幼鼠结肠白细胞介素-17A (IL-17A)水平的影响,探讨益生菌对Th17细胞的调节作用。方法 36只健康雄性SD幼鼠随机分4组:空白对照组、LGG对照组各8只,IBD组、IBD-LGG组各10只。利用2,4,6-三硝基苯磺酸(TNBS)诱导幼鼠IBD模型,观察一般状况、IBD疾病活动指数评分。第8天处死所有幼鼠,留取结肠标本,观察病理改变并采用免疫组织化学法测定结肠组织IL-17A的表达。结果相比两对照组,IBD组、IBD-LGG组幼鼠一般状态差,IBD-LGG组便性状及隐血较IBD组缓解;IBD组、IBD-LGG组幼鼠结肠组织均见炎症改变,但IBD-LGG组较轻。IBD-LGG组DAI评分、IL-17A水平均低于IBD组,差异有统计学意义(Ps0.05)。结论益生菌可减轻IBD幼鼠肠道炎症,其机制可能与益生菌调节Th17细胞进而调控IL-17A表达有关。     

A primary dysregulation in the immunoregulatory role of the intestinal mucosal epithelial cell in inflammatory bowel disease pathogenesis? Biology of inflammatory response as tissue pattern entities in Crohn's versus ulcerative colitis

Within a framework of dual involvement of mucosa and submucosa on the one hand, and of the muscularis propria of the bowel wall on the other, it might be valid to consider involvement of the vascular supply as the essential means in itself of not only causing the morphologic lesions in inflammatory bowel disease, but also especially in accounting for persisting patterns of inflammatory response both in ulcerative colitis and in Crohn's disease. Inflammatory bowel disease as a group constitutes a spectrum of biologic and pathobiologic manifestations in terms not only of inflammatory involvement of the bowel wall but also in terms of how the bowel in its turn deals with inflammation as a pathologic lesion in its own right. Parameters of inflammatory bowel activity transcend simple concepts of etiology and pathogenesis as applicable to category disorders such as infections or bowel ischemia. Indeed, the strictly characterized initiation of the inflammatory bowel response as a function of defective regulation of the antigenicity of the luminal contents on the one hand, and on interactions between nitric oxide and free oxygen radicals on the other, might help determine a persistence of tissue damage in inflammatory bowel disease that is either relapsing/remitting or chronic in progression. In a final analysis, perhaps, there might be involved a single central form of pathway induction of dysregulated immune reactivity arising from an early disturbance in activation patterns as induced by the onset of luminal antigenicity at an early or specific-stage, further characterized perhaps by specific forms of intestinal epithelial defects of the bowel mucosa in patients subsequently developing inflammatory bowel disease. Specific genetic markers for disease susceptibility and for therapeutic responsiveness are particularly of interest. The Nucleotide binding oligomerization Domain 2 (NOD2) would recognize microbial lipopolysaccharide or else mark systemic responses to pathogens that are pathogenic to evolving inflammatory bowel disease.     

Therapeutic effects of lentinan on inflammatory bowel disease and colitis‐associated cancer

In this study, we investigated the therapeutic potential of lentinan in mouse models of inflammatory bowel disease (IBD) and colitis‐associated cancer (CAC). Lentinan decreased the disease activity index and macroscopic and microscopic colon tissue damage in dextran sulphate sodium (DSS)‐induced or TNBS‐induced models of colitis. High‐dose lentinan was more effective than salicylazosulfapyridine in the mouse models of colitis. Lentinan decreased the number of tumours, inflammatory cell infiltration, atypical hyperplasia and nuclear atypia in azoxymethane/DSS‐induced CAC model. It also decreased the expression of pro‐inflammatory cytokines, such as IL‐13 and CD30L, in IBD and CAC model mice possibly by inhibiting Toll‐like receptor 4 (TLR4)/NF‐κB signalling and the expression of colon cancer markers, such as carcinoembryonic antigen, cytokeratin 8, CK18 and p53, in CAC model mice. In addition, lentinan restored the intestinal bacterial microbiotal community structure in IBD model mice. Thus, it shows therapeutic potential in IBD and CAC model mice possibly by inhibiting TLR4/NF‐κB signalling‐mediated inflammatory responses and disruption of the intestinal microbiotal structure.     

Lower prevalence of Helicobacter pylori infection in patients with inflammatory bowel disease but not with chronic obstructive pulmonary disease - antibiotic use in the history does not play a significant role

BACKGROUND: Patients with inflammatory bowel disease have lower prevalence of Helicobacter pylori infection, but the exact reason for this is not yet clear. AIM: To examine whether the antibiotics frequently used in inflammatory bowel disease are responsible for the lower prevalence of H. pylori infection. Patients with chronic obstructive pulmonary disease on prolonged previous antibiotic therapy were used for comparison. METHODS: Presence/absence of H. pylori infection was detected by a (13)C-urea breath test in 133 patients with inflammatory bowel disease (82 ulcerative colitis, and 51 Crohn's disease) and compared with that of 135 patients with chronic obstructive pulmonary disease and with two age-matched control groups (200 patients each). Primary disease location, duration of disease and detailed analysis of previous and current medication (dose and duration of antibiotics, steroids, 5-aminosalicylic acid) were analysed in each cases. RESULTS: Seventeen of the 133 patients with inflammatory bowel disease [12.2% (10/82) of ulcerative colitis and 13.7% (7/51) of Crohn's disease] and 90/135 patients with chronic obstructive pulmonary disease (66.7%) were positive for H. pylori. A total of 78/200 (39%) for the inflammatory-bowel-disease-group-matched controls and 110/210 (55%) for the chronic-obstructive-pulmonary-disease-matched controls were positive for H. pylori. The history of any antibiotic or steroid therapy had no influence on H. pylori status of patients with inflammatory bowel disease. CONCLUSION: The prevalence of H. pylori compared to the age-matched controls is significantly lower in patients with inflammatory bowel disease but not in those with chronic obstructive pulmonary disease. Antibiotic use is not responsible for the lower prevalence of H. pylori infection in patients with inflammatory bowel disease.     

Antioxidants and mucosa protectives: realistic therapeutic options in inflammatory bowel disease?

Oxidative damage is involved in the pathogenic process of idiopathic chronic inflammatory bowel disease. Although specific intervention in the oxidative cascade showed promising results in animal models and preliminary patient trials, the clinical efficacy of antioxidants still has to be established. Mucosa protection, for example by dietary fatty acids, seems to attenuate the intestinal inflammatory process as well but awaits definite clinical proof for the treatment of inflammatory bowel disease.     

The Complement Anaphylatoxin C3a Receptor (C3aR) Contributes to the Inflammatory Response in Dextran Sulfate Sodium (DSS)-Induced Colitis in Mice

Inflammatory bowel diseases are a critical public health issue, and as treatment options remain limited, there is a need to unravel the underlying pathomechanisms in order to identify new therapeutic targets. Complement activation was found in patients suffering from inflammatory bowel disease, and the complement anaphylatoxin C5a and its receptor C5aR have been implicated in disease pathogenesis in animal models of bowel inflammation. To further characterize complement-related pathomechanisms in inflammatory bowel disease, we have investigated the role of the anaphylatoxin C3a receptor in acute dextran sulfate sodium-induced colitis in mice. For this, colitis was induced in C3a receptor-deficient BALB/c and C57BL/6 mice, and disease severity was evaluated by clinical and histological examination, and by measuring the mRNA expression or protein levels of inflammatory mediators in the tissue. C3a receptor deficiency was partially protective in BALB/c mice, which had significantly reduced weight loss, clinical and histological scores, colon shortening, and CXCL-1/KC mRNA, myeloperoxidase and interleukin-6 tissue levels compared to the corresponding wild type mice. In C57BL/6 mice the differences between wild type and C3a receptor-deficient animals were much smaller and reached no significance. Our data demonstrate that the contribution of C3a receptor to disease pathogenesis and severity of dextran sulfate sodium-induced colitis in mice depends on the genetic background. Further studies will be required to clarify whether targeting of C3a receptor, possibly in combination with C5a receptor, might be considered as a therapeutic strategy for inflammatory bowel disease.     

The role of obesity in inflammatory bowel disease

In just over a generation overweight and obesity has become a worldwide health concern. The ramifications for this on future health care costs and longevity are consequent, whilst increased adiposity is a harbinger for diabetes, kidney and bone failure, and cancer. An area of intense interest where the role of adiposity is avidly discussed is in inflammatory bowel disease (IBD), which presents mainly as Crohn's disease (CD) and ulcerative colitis (UC). Studies in patients associating IBD with a western diet are divergent. Nevertheless, elegant studies have found gene polymorphisms in humans that in murine models parallel the inflammatory and gut microbiome changes seen in IBD patients. However, an area not to be ignored are the alterations in adipocyte function with ensuing adiposity, in particular and a focus of this review, the dysregulation of the levels of adipocytokines such as leptin and adiponectin. Herein, we present and discuss the known influences of a western diet on IBD in patients and rodent models and how adipocytokines could influence the IBD disease process.     

Immunomodulatory benefits of cyclosporine A in inflammatory bowel disease

Etiopathogenesis of mucosal inflammation in inflammatory bowel disease remains a complex and enigmatic field; various factors (genetic, environmental and microbial) trigger an event that activates intestinal immune and nonimmune systems culminating in inflammation and tissue injury. Specifically, both innate and adaptive immune systems seem to play important roles in the pathophysiology of this disease. Cyclosporine A represents a macrolide immune modulator with primary inhibitory effects on T helper lymphocyte production of interleukin-2, and other cytokines leading to altered T-lymphocyte and B-lymphocyte function. The diversity of its therapeutic outcome reported in inflammatory bowel disease may be due to the intricate immuno-pathogenic profile of the disease and the variety of the applied dose-dependent courses of therapy. Cyclosporine A exerts additional actions on other components of the inflammatory infiltrate, including neutrophils and mast cells, thereby appearing to be a multi-dynamic therapeutic approach, although with potential drawbacks, that may be applied alone or combined with other immunomodulatory agents in inflammatory bowel disease patients. Because cyclosporine A induces apoptosis of T-lymphocytes responsible for perpetuation of the chronic inflammatory process in the disease with potential tumorigenic effect, it may exert a further inhibitory effect on cancer development in inflammatory bowel disease patients, and can be combined with other relative agents, such as rapamycin, which also promotes T-lymphocyte apoptosis. Therefore, recently established multifactorial action of cyclosporine A in relation to the pathogenesis of the disease can open new horizons for prospective, controlled trials in large cohorts, aiming to emphasize cyclosporine A's potential.     

金属硫蛋白与医学

金属硫蛋白是一类富含巯基与金属的内源性蛋白,化学结构极为特殊,该蛋白的为具有很强的诱导性,金属,糖皮质激素,某些化的和应激状态均可诱导其生成,在炎症等应激过程中ＩＬ－６可能为诱导者。在不降低疗效的情况下该蛋白可解除抗肿瘤药物ＣＤＤＰ的抗药性密切有关,金属硫蛋白与炎性肠病,妊娠中毒症,肝脏疾患等多种疾病密切相关。     

IL‐35 recombinant protein reverses inflammatory bowel disease and psoriasis through regulation of inflammatory cytokines and immune cells

Interleukin‐35 (IL‐35), a member of the IL‐12 family, functions as a new anti‐inflammatory factor involved in arthritis, psoriasis, inflammatory bowel disease (IBD) and other immune diseases. Although IL‐35 can significantly prevent the development of inflammation in many diseases, there have been no early studies accounting for the role of IL‐35 recombinant protein in IBD and psoriasis. In this study, we assessed the therapeutic potential of IL‐35 recombinant protein in three well‐known mouse models: the dextransulfate sodium (DSS)‐induced colitis mouse model, the keratin14 (K14)‐vascular endothelial growth factor A (VEGF‐A)‐transgenic (Tg) psoriasis mouse model and the imiquimod (IMQ)‐induced psoriasis mouse model. Our results indicated that IL‐35 recombinant protein can slow down the pathologic process in DSS‐induced acute colitis mouse model by decreasing the infiltrations of macrophages, CD4⁺T and CD8⁺T cells and by promoting the infiltration of Treg cells. Further analysis demonstrated that IL‐35 recombinant protein may regulate inflammation through promoting the secretion of IL‐10 and inhibiting the expression of pro‐inflammatory cytokines such as IL‐6, TNF‐α and IL‐17 in acute colitis model. In addition, lower dose of IL‐35 recombinant protein could achieve long‐term treatment effects as TNF‐α monoclonal antibody did in the psoriasis mouse. In summary, the remarkable therapeutic effects of IL‐35 recombinant protein in acute colitis and psoriasis mouse models indicated that IL‐35 recombinant protein had a variety of anti‐inflammatory effects and was expected to become an effective candidate drug for the treatment of inflammatory diseases.