右旋葡聚糖硫酸钠小鼠溃疡性结肠炎动物模型建立方法探讨

溃疡性结肠炎（UC）的病因和发病机制尚不十分清楚，建立适当的结肠炎动物模型对于研究其病因和发病机制具有重要意义。目的：探讨右旋葡聚糖硫酸钠（DSS）诱发的小鼠UC动物模型的建立方法。方法：将18只雄性BALB／C小鼠随机分为3组，实验组中一组饮用5％DSS溶液7天诱发急性结肠炎，另一组饮用5％DSS溶液7天后继续饮用蒸馏水14天诱发慢性结肠炎；对照组饮用蒸馏水。观察小鼠每日的体重、大便性状和隐血情况以及结肠大体形态和组织病理学改变。结果：饮用5％DSS溶液7天，BALB／C小鼠可发生腹泻、血便等症状，全结肠表现为以隐窝破坏为特征的多灶性小溃疡，伴中性粒细胞为主的急性炎症细胞浸润。停止饮用5％DSS溶液14天后，BALB／C小鼠的腹泻、血便等症状消失，全结肠表现为更局限的灶性小溃疡伴邻近上皮细胞再生、修复，以及突出的隐窝扭曲变形伴以淋巴、单核细胞为主的慢性炎症细胞浸润。急、慢性期肠道病变均以远段结肠为重。结论：单次使用DSS诱发的小鼠急、慢性结肠炎是一种较理想的UC动物模型，可作为研究UC发病机制和药物治疗较理想的工具。     

Plasma levels of interleukin-18 and interleukin-12 in Plasmodium falciparum malaria

Interleukin (IL)-18 produced primarily by mononuclear phagocytes synergizes with IL-12 for interferon-gamma production from T, B and natural killer cells. It has been also demonstrated that, in Plasmodium falciparum malaria, IL-18 could have an immunoregulatory function. The aim of this study was to detect the plasma levels of IL-12 and IL-18, using an enzyme-linked immunosorbent assay, in 105 African children affected by mild and severe Plasmodium falciparum malaria to correlate the production of these cytokines with the severity of the disease. The levels of IL-18 and IL-12 were higher (25.7 +/- 7.6 pg/ml and 17.1 +/- 7.8 pg/ml, respectively) in children with mild malaria than in children with a severe form of the disease (21.5 +/- 10 pg/ml and 13.2 +/- 5.5 pg/ml, respectively). A positive correlation was observed between IL-18 and IL-12. This finding suggests that the production of these two cytokines (IL-18 and IL 12) may be coregulated and both have an immunoregulatory effect on the immune response in Plasmodium falciparum infection.     

白细胞介素-12及白细胞介素-18致小鼠脾虚的研究

目的：观察白细胞介素-12(IL-12)及白细胞介素-18(IL-18)能否导致脾虚，研究脾虚与IL-12及IL-18的关系。方法：60只小鼠随机分为脾虚组、IL组和对照组。采用利血平制作脾虚动物模型组，IL组每日腹腔注射IL-12、IL-18。采用免疫分析技术检测3组小鼠血浆IL-12、IL-18的含量。结果：IL-12及IL-18引起明显脾虚证的表现，与利血平制作脾虚动物模型组相似。脾虚组和IL组血浆IL-12、IL-18的含量明显升高。结论：IL-12、IL-18可导致脾虚，血浆IL-12、IL-18的含量可作为脾虚证的实验室参考指标。     

发酵黑麦糠对右旋葡聚糖硫酸钠诱导的小鼠结肠炎的保护作用

溃疡性结肠炎是一种病因和发病机制尚不明确的结肠黏膜和黏膜下层慢性炎症。体内外试实验表明，发酵黑麦糠可抑制幽门螺杆菌(H．pylori)对胃黏膜的黏附作用，从而保护小鼠免受H．pylori感染。目的：明确发酵黑麦糠是否对右旋葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎有保护作用。方法：以3．5％DSS诱导小鼠产生结肠炎，部分小鼠同时加用发酵黑麦糠，观察结肠炎小鼠的临床表现(体重下降情况、直肠出血情况和有无腹泻)和结肠黏膜的组织学改变。结果：DSS 发酵黑麦糠组小鼠结肠炎起病晚、症状轻，在观察期内无一例出现腹泻；组织学检查和评分结果亦显示该组小鼠结肠黏膜炎症较DSS组轻。结论：发酵黑麦糠对DSS诱导的小鼠结肠炎有保护作用，其具体作用机制尚有待进一步明确。     

Inhibition of Selectin Function and Leukocyte Rolling Protects Against Dextran Sodium Sulfate-Induced Murine Colitis

Background: The selectin family of adhesion molecules (P-, E- and L-selectin) plays an important role in inflammatory reactions by mediating interactions between leukocytes and activated endothelial cells. However, a recent study using gene-targeted mice has suggested that adhesion molecules (P- and Eselectin and ICAM-1) may not be relevant targets in intestinal inflammation. The objective of the present study was to re-evaluate the potential role of selectins in experimental colitis in wild-type mice using the polysaccharide fucoidan, which inhibits the function of P- and L-selectin. Methods: For this purpose, Balb/c mice were exposed to 5% dextran sodium sulfate (DSS) in the drinking water for 5 days with and without daily administration of fucoidan (25 mg/kg, i.v.). In separate experiments, the effect of fucoidan on leukocyte-endothelium interactions was examined by use of intravital microscopy. Results:     

丹参对右旋葡聚糖硫酸钠诱导小鼠结肠炎的疗效观察

目的 :评价丹参预防及治疗右旋葡聚糖硫酸钠 (DSS)结肠炎小鼠的有效性。方法 :2 0只正常小鼠随机分为两组 ,饮用 DSS7d,同时预防组用丹参 ,对照组用 0 .85 %氯化钠溶液。另 2 0只 DSS诱导的结肠炎小鼠随机分为两组 ,治疗组用丹参 ,对照组用 0 .85 %氯化钠溶液 7d。用疾病活动指数 (DAI)、组织学评分和马休斯猩红蓝(MSB)纤维素染色检测微血栓以评价疗效。结果 :丹参在预防组部分降低微血栓的形成 ,对照组 10例有 6例微血栓阳性 ,预防组 3例阳性。丹参治疗组与对照组的 DAI、直肠、横结肠组织学评分分别为 0 .4 5、0 .4 8(P>0 .0 5 ) ,1.36、1.76 (P<0 .0 5 ) ,1.35、1.6 0 (P<0 .0 5 )。结论 :丹参可能部分抑制微血栓形成和减轻 DSS结肠炎小鼠结肠炎症 ,提示丹参用于溃疡性结肠炎治疗也可能有效。     

Fructo-oligosaccharide Reduces Inflammation in a Dextran Sodium Sulphate Mouse Model of Colitis

Fructo-oligosaccharide (FOS) is a prebiotic that stimulates the colonic growth of bifidobacteria to promote intestinal health. This study assessed whether FOS can reduce intestinal damage associated with ulcerative colitis and accelerate recovery in a mouse model. C57BL/6 mice received 2% dextran sulphate sodium for 7 days (days 8–14). FOS (1.5 g/mL) treatment was administered twice daily (n=10/group): before and during colitis (days 1–14); during colitis (days 10–14); and during colitis and the recovery period (days 10–19). Disease activity was scored daily and colonic damage was assessed by histological analysis. FOS treatment significantly reduced disease activity and damage in the distal colon (P < .05). Treatment with FOS (days 10–14) had increased crypt depth (116±6 μm) compared to water treatment (90±4 μm, P < .05). FOS treatment (days 10–19) produced a faster recovery from damage with increased crypt depth and crypt area. These results demonstrate the protective effect of FOS treatment.     

Probiotics and Blueberry Attenuate the Severity of Dextran Sulfate Sodium (DSS)-Induced Colitis

We studied the anti-inflammatory properties of probiotic strains and blueberry in a colitis model. The disease activity index (DAI) was significantly lower on days 9 and 10 in all groups compared to the colitis control. Myeloperoxidase (MPO) and bacterial translocation to the liver and to the mesenteric lymph nodes (MLN) decreased significantly in all groups compared to colitis control. Cecal Enterobacteriaceae count decreased significantly in blueberry with and without probiotics compared to the other groups. Lactobacillus plantarum reisolated from the cecal content in the presence of blueberry, contrary to Lactobacillus fermentum. Colonic MDA decreased significantly in all groups, except the L. fermentum group, compared to the colitis control. The cecal concentration of acetic, propionic, and butyricbutyric acid was significantly higher in the L. plantarum group, while the L. fermentum group yielded the highest concentration of lactic acid compared with all other groups. Lactobacillus plantarum DSM 15313, Lactobacillus fermentum 35D, and blueberry alone and in combination improve the DAI, reduce bacterial translocation, and reduce inflammation.     

小鼠葡聚糖硫酸钠结肠炎模型结肠黏膜中5-羟色胺及其转运蛋白的变化和意义

背景：小鼠葡聚糖硫酸钠（DSS）结肠炎模型的组织学特点与人类溃疡性结肠炎（UC）类似。5-羟色胺（5-HT）是胃肠道重要的神经递质，其在UC中的变化和意义鲜见报道。目的：测定小鼠DSS结肠炎模型结肠黏膜中5-HT和5-HT转运蛋白（SERT）的变化，探讨UC是否与5-HT信号转导失衡有关。方法：36只C57BL／6小鼠随机分为急、慢性DSS结肠炎组和正常对照组，以免疫组化方法检测5-HT和SERT的表达，以免疫荧光技术检测SERT的免疫反应性。结果：与正常对照组相比，急、慢性DSS结肠炎组远段和近段结肠黏膜5-HT平均灰度值显著降低（含量增加，P〈0．05），远段结肠黏膜SERT平均灰度值显著增高（含量降低，P〈0．05）。正常对照组、慢性和急性DSS结肠炎组结肠黏膜依次发出耀眼、明亮和微弱的黄绿色荧光，提示DSS结肠炎组SERT免疫反应性减弱。结论：DSS诱导的结肠炎促使小鼠结肠黏膜中5-HT含量和SERT的表达发生变化，5-HT和SERT可能参与了UC发生的病理生理机制。     

Keratinocyte Growth Factor Ameliorates Dextran Sodium Sulfate Colitis in Mice

Keratinocyte growth factor (KGF) is emerging asan important mediator of mucosal defense and repair inthe colon. The aim of the present study was to evaluateand further characterize the effects of exogenous KGF administration utilizing the dextran sodiumsulfate (DSS) model of colitis in mice. Colitis wasinduced via oral administration of DSS (5 g/100 ml) toBalb/c mice for eight days. Intraperitonealadministration of KGF (5 mg/kg, once daily) or vehicle (VEH)was initiated 1 hr prior to the induction of the colitis(N = 10, each group). Mucosal injury of the entire colonwas histologically assessed and graded. An approximately fourfold reduction in the cryptdamage score was noted in the KGF group when compared tocontrols (VEH) (2.8 ± 1.03 and 11.4 ±0.78, respectively). The significant reduction ofmucosal injury in KGF treated mice confirms that KGF isa key mediator maintaining the integrity of the colonicmucosa.     

葡聚糖硫酸钠自由饮用与定量灌胃诱导小鼠急性结肠炎模型的比较研究

背景：自由饮用葡聚糖硫酸钠（DSS）诱导的鼠类急性结肠炎模型均一性欠佳,动物死亡率较高。目的：评估2%DSS自由饮用或定量灌胃诱导的小鼠急性结肠炎模型模拟人类溃疡性结肠炎（UC）的效果和均一性。方法：予ICR小鼠2%DSS自由饮用或定量灌胃建立急性结肠炎模型,检测并比较正常对照组和两组模型小鼠的症状出现时间和频率、疾病活动指数（DAI）、DSS消耗量、死亡率、结肠长度、结肠损伤大体评分（MACD）、结肠组织病理学表现以及外周血白细胞计数和分类。结果：两组模型小鼠均出现类似人类UC的症状和组织病理学改变,结肠显著短缩,DAI、MACD以及外周血白细胞计数和中性粒细胞百分比显著高于正常对照组。与DSS自由饮用组相比,DSS定量灌胃组小鼠症状出现时间更为一致,症状出现率显著增高,动物死亡率和DSS消耗量显著减低。结论：2%DSS定量灌胃诱导的小鼠急性结肠炎模型能较稳定地模拟人类UC,均一性高,动物死亡率低。     

慢性心力衰竭患者血清白细胞介素-12及白细胞介素-18水平及意义

目的:检测慢性心力衰竭患者(CHF)外周血白细胞介素(IL)-12和IL-18的水平并探讨其意义.方法:CHF患者72例,按照NYHA心功能分级,分为HF1(NYHAⅡ～Ⅲ级),HF2组(NYHA Ⅳ级).按照病因分类,分为缺血性心脏病(IHD)组(43例)和非缺血性心脏病(NIHD)组(29例).选取29例健康志愿者作为正常对照组(C组).采用双抗体夹心酶联免疫吸附法(ELISA)检测血清中IL-12和IL-18的水平,用彩色多普勒超声诊断仪测定左室舒张末期内径(LVEDd)和左室射血分数(LVEF).结果:HFI、HF2、IHD和NIHD组IL-12的水平显著升高,分别为(34.56±19.10),(35.12±15.18),(31.18±7.80),(32.73±11.54)ng/L,与C组(28.73±13.43)ng/L比较,均差异有统计学意义(P＜0.05),IL-18的水平分别为(39.50±5.66),(46.76±12.66),(40.58±13.10),(42.53±6.16)ng/L,明显高于C组(31.82±5.79)ng/L,均差异有统计学意义(P＜0.05或P＜0.01),HF2组血清IL-18水平显著高于HF1组(P＜0.05).与C组比较,HF1、HF2、IHD和NIHD组的LVEDd明显增大.LVEF显著降低.IL-12水平与LVEDd呈低度正相关(P＜0.01),IL-18水平与LVEDd呈中度正相关(P＜0.01),与LVEF呈中度负相关.结论:IL-12和IL-18在CHF患者血清中表达升高;IL-12的水平与患者基础病因和心功能均无明显相关性;IL-18的水平与CHF的基础病因无明显相关性,但与患者的CHF程度呈正相关.     

SLE患者血清及外周血单个核细胞中IL-12、IL-18的检测及临床意义

目的探讨系统性红斑狼疮（SLE）患者IL-12、IL-18的表达及与疾病活动的关系。方法应用ELISA法以及半定量RT-PCR法分别测定SLE活动组、缓解组和对照组血清IL-12、IL-18水平以及外周血单个核细胞中的IL-12 mRNA和IL-18 mRNA的表达。结果SLE活动组IL-18 mRNA、IL-18的表达较SLE缓解组和正常对照组明显升高（P均〈0.05）。血清IL-18水平与IL-12水平呈负相关（r=-0.406,P=0.019）。SLE患者SLE疾病活动指数与IL-12、IL-12 mRNA呈负相关（r=-0.617,P〈0.05;r=-0.512,P〈0.05）,与IL-18、IL-18 mRNA呈正相关性（r=0.817,P〈0.05;r=0.806,P〈0.01）。结论SLE活动组患者中存在IL-18异常高表达和IL-12异常低表达,二者之间呈负相关,在SLE发病和发展中可能起不同作用。     

葡聚糖硫酸钠加乙酸复合法制作结肠炎大鼠模型的初探

背景:动物模型对阐明溃疡性结肠炎(UC)的发病机制和评价治疗效果具有重要价值,葡聚糖硫酸钠(DSS)因价格较昂贵而使其在结肠炎模型制作中的应用受到限制。目的:探讨DSS加乙酸复合法制作结肠炎大鼠模型的可行性和优缺点。方法:12只Sprague-Dawley大鼠随机分为对照组和模型组。模型组大鼠给予3%DSS溶液自由饮用7 d,第8 d和第16 d,以8%乙酸2 ml灌肠;对照组大鼠予等体积纯净水饮用和等体积纯净水灌肠。实验第24 d处死大鼠,行疾病活动指数(DAI)评分、结肠黏膜损伤大体评分和组织病理学评分。结果:与对照组相比,模型组大鼠DAI评分(0.87±0.61对-1.92±1.03)、结肠黏膜损伤大体评分(3.00±0.71对0.00±0.00)和组织病理学评分(4.60±1.52对0.00±0.00)均显著升高(P0.05)。结论:DSS加乙酸复合法能较好地复制出具备人类UC特点的大鼠模型,且成本相对较低。     

LADA患者血清IL-12、IL-18及血淋巴细胞亚群研究

与2型糖尿病组及正常对照组相比,成人隐匿性自身免疫糖尿病(LADA)组外周血淋巴细胞亚群有明显变化,白细胞介素12(IL12)、白细胞介素18(IL18)水平亦升高。LADA存在明显细胞免疫紊乱,并与胰岛功能密切相关。     

葡聚糖硫酸钠诱导大鼠结肠炎发病机制的研究

背景:葡聚糖硫酸钠(DSS)可诱导大鼠结肠炎,但其发病机制尚不完全清楚。目的:探讨DSS诱导大鼠结肠炎的发病机制。方法:16只健康雄性Sprague-Dawley大鼠随机分成DSS模型组(10只)和正常对照组(6只),DSS模型组大鼠自由饮用2％DSS溶液8天,正常对照组大鼠正常饮水。于第9天处死大鼠,采用酶联免疫吸附测定(ELISA)方法测定血清肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6水平,采用免疫组化方法测定结肠黏膜细胞间黏附分子(ICAM)-1的表达和核因子(NF)-κB活性。结果:DSS模型组大鼠出现腹泻、便血以及结肠黏膜糜烂或溃疡,血清TNF-α和IL-6水平以及结肠黏膜ICAM-1表达和NF-κB活性均较正常对照组显著增高(P<0.01)。结论:在DSS诱导的大鼠结肠炎中,DSS可能通过活化NF-κB使TNF-IL-6和ICAM-1生成增加,导致结肠黏膜炎性损害和腹泻、便血。     

慢性乙型肝炎患者血清IL-12、IL-18水平和外周血单个核细胞FOXp3基因水平研究*

目的 探讨慢性乙型肝炎（CHB）患者血清白细胞介素-12（IL-12）、IL-18和外周血单个核细胞叉头蛋白P3（Foxp3）基因水平变化及其临床意义。方法 2015年7月~2018年3月我院收治的CHB患者78例和同期健康体检者25例,采用ELISA法检测血清IL-12和IL-18水平,分离外周血单个核细胞,采用RT-PCR法检测细胞Foxp3 mRNA水平。结果 在78例CHB患者中,经肝组织活检诊断G1组21例,G2组24例,G3组19例,G4组14例;CHB患者血清ALT和AST水平分别为（278.3±89.4）U/L和（305.3±84.6） U/L,显著高于健康人的[（25.3±7.6） U/L和（20.3±6.5） U/L,P<0.05];G4组血清ALT和AST水平分别为（563.5±132.4） U/L和 （513.3±102.5） U/L,显著高于G1组[（113.2±67.4）U/L和（73.5±25.3） U/L]或G2组[（153.6±45.3） U/L和（113.8±35.2） U/L]或G3组[（240.5±56.6） U/L和（156.7±51.2） U/L,P<0.05];CHB患者血清IL-12和IL-18水平及单个核细胞Foxp3 mRNA水平分别为（198.3±19.6） pg/ml、（408.6±91.2） pg/ml及（4.5±0.7）,显著高于健康人[（64.5±10.9） pg/ml、（127.3±15.7） pg/ml、（3.3±0.4）,P<0.05];G4组血清IL-12、IL-18及Foxp3基因水平分别为（237.5±23.6） pg/ml、（431.5±106.3） pg/ml、（5.1±0.3）],显著高于G1组[（146.3±15.2） pg/ml、（390.2±90.3） pg/ml、（3.7±0.6）] 或G2组[（185.2±13.6） pg/ml、（403.6±93.2） pg/ml、（3.9±0.5） pg/ml]或G3组[（203.2±18.3）pg/ml、（414.3±105.4） pg/ml、（4.3±0.7）,P<0.05]。结论 CHB患者外周血单个核细胞Foxp3基因及血清IL-12和IL-18水平显著升高,且肝组织炎症活动显著者,其水平更高,提示其参与了CHB的发病过程。及时检测这些指标可能对监测病情变化和判断治疗应答有帮助。     

Regulatory effects of IL-12 and IL-18 on Onchocerca volvulus- and Entamoeba histolytica-specific cellular reactivity and cytokine profiles

In the present study, the cytokines interleukin (IL)-12 and IL-18 were evaluated for their capacity to modulate and to re-direct in vitro parasite antigen-specific cellular responsiveness in patients exposed to Onchocerca volvulus and Entamoeba histolytica infection. We found that IL-18 was highly capable of reducing parasite antigen-induced IL-10 production by PBMC. In contrast, addition or neutralization of IL-12, also in combination with IL-18 and the interferon-gamma-inducible chemokine IP-10 did not affect IL-10 production. Interestingly, the highest IL-10 levels were measured when IL-18 and IP-10 were both neutralized. Although having no effect on IL-10, IL-12 strongly promoted spontaneous and parasite antigen-driven IFN-gamma production by PBMC, whereas IL-18 was only moderately affecting IFN-gamma release by PBMC re-stimulated with E. histolytica- or O. volvulus-specific antigens. Both IL-12 and IL-18 diminished the cellular production of IL-13, and a synergistic effect was observed when the cytokines were combined. Likewise, neutralization of IL-12 enhanced Entamoeba and Onchocerca antigen-driven IL-13 production, but no further increase of IL-13 was observed, when anti-IL-12 and anti-IL-18 were used together. This study disclosed that IL-18 will significantly down-regulate parasite-specific IL-10 production, whereas IL-12 induced IFN-gamma and inhibited IL-13 production by PBMC from humans exposed to O. volvulus and E. histolytica. Such selective immune-regulatory capacity of IL-12 and IL-18 may comprise an important tool to re-direct polarized cytokine responses towards a balanced Th1/Th2 cytokine profile, which may prevent pathology and promote immunity against helminth and protozoan parasite infections.