雷公藤多甙对(口恶)唑酮诱导小鼠肠炎模型脾淋巴细胞因子的作用及其机制

目的　观察小鼠肠炎模型体外实验中 ,雷公藤多甙 (MGT)对唑酮 (oxazolone,OXZ)诱导的小鼠肠炎模型中脾脏淋巴细胞细胞因子分泌类型的影响 ,从免疫学、分子生物学角度探讨MGT治疗炎症性肠病 (IBD)的作用机制。方法　采用SJL/J小鼠 ,经直肠注入OXZ制成IBD模型 ;3d后将小鼠处死 ,即刻取出脾脏 ,收集脾细胞 ,将MGT(0 .1和 0 .0 1mg/ml两个浓度 )分别加入培养的淋巴细胞液中 ,行ELISA检测白介素 4 (IL 4 )和γ 干扰素 (IFN γ)。结果　 1.MGT对IFN γ生成的影响 :(1)正常对照组 :未加入MGT组 (空白对照 )的IFN γ为 (1.2 4± 0 .13) pg/ml;0 .0 1mgMGT组为(0 .97± 0 .2 6 )pg/ml;0 .1mgMGT组为 (0 .87± 0 .18) pg/ml;(P <0 .0 2 ) ;(2 )OXZ肠炎模型组 :与正常对照组比较 ,未加入MGT组 (空白对照 )IFN γ显著降低 [(0 .6 5± 0 .0 8) pg/ml比 (1.2 4±0 .13) pg/ml,P <0 .0 1],0 .0 1mgMGT组和 0 .1mgMGT组IFN γ均低于空白对照 [分别为 (0 .4 7± 0 .0 5 ) pg/ml;(0 .4 6± 0 .0 9) pg/ml],但差异无显著性。 2 .MGT对IL 4生成的影响 :(1)正常对照组 :未加入MGT组 (空白对照 )的IL 4为 (5 .6 5± 0 .4 8) pg/ml;MGT有显著抑制作用 [0 .0 1mg/mlMGT组为(4.97± 0 .38) pg/ml;0 .1mgMGT组为 (3.98± 0 .32     

雷公藤多甙对恶唑酮诱导小鼠肠炎模型脾淋巴细胞因子的作用及其机制

目的观察小鼠肠炎模型体外实验中,雷公藤多甙(MGT)对唑酮(oxazolone,OXZ)诱导的小鼠肠炎模型中脾脏淋巴细胞细胞因子分泌类型的影响,从免疫学、分子生物学角度探讨MGT治疗炎症性肠病(IBD)的作用机制.方法采用SJL/J小鼠,经直肠注入OXZ制成IBD模型;3d后将小鼠处死,即刻取出脾脏,收集脾细胞,将MGT(0.1和0.01 mg/ml两个浓度)分别加入培养的淋巴细胞液中,行FILISA检测白介素-4(IL-4)和γ-干扰素(IFN-γ).结果1.MGT对IFN-γ生成的影响(1)正常对照组未加入MGT组(空白对照)的IFN-γ为(1 24±0 13)pg/ml;0.01mg MGT组为(0.97±0 26)pg/ml;0.1mg MGT组为(0 87±0 18)pg/ml;(P＜0.02);(2)OXZ肠炎模型组与正常对照组比较,未加入MGT组(空白对照)IFN-γ显著降低[(0 65±0 08)pg/ml比(1.24±0.13)pg/ml,P＜0.01],0.01mg MGT组和0.1mg MGT组IFN-γ均低于空白对照[分别为(0.47±0.05)pg/ml;(0 46±0 09)pg/ml],但差异无显著性.2.MGT对IL-4生成的影响(1)正常对照组未加入MGT组(空白对照)的IL4为(5.65±0.48)pg/ml;MGT有显著抑制作用[0.01 mg/ml MGT组为(4.97±0.38)pg/ml;0.1 mgMGT组为(3.98±0.32)pg/ml;P＜0.01];(2)0XZ肠炎模型组与对照组比较,未加入MGT组(空白对照)IL-4显著升高[(7.83±0.69)pg/ml比(5.65±0.48)pg/ml,P<0.01];0.01mg MGT组(7.07±0.47)pg/ml和0.1 mg MGT组(6.35±0.48)pg/m1与空白对照组比较,差异有显著性(P<0.01).结论雷公藤既可抑制IFN-γ(Th1)生成,亦可抑制IL-4(Th2)分泌.由于OXZ诱导的小鼠实验性肠炎模型系以1L-4过量生成为主的Th2型免疫反应,由此可以推测MGT治疗溃疡性结肠炎的机制可能与其抑制IL-4(Th2)生成有关.     

吡格列酮对噁唑酮诱导的小鼠实验性结肠炎模型的影响

目的　观察吡格列酮对唑酮结肠炎的干预 ,探讨其是否通过Fas Fas配体 (FasL)途径促进细胞凋亡参与免疫调节作用。方法　经唑酮诱导成功的BALb/c小鼠 ,每组 6只 ,第 1组于灌肠后3d分离结肠固有层单个核细胞 (LPMC) ,并在体外用吡格列酮 (40 μmol/L)处理 2 4h ,采用annexin V FITC试验法分析凋亡细胞的百分率 ,采用流式细胞术检测Fas及FasL表达 ;并没未处理的正常小鼠组。第 2组 (对照组 )和第 3组 (实验组 )于灌肠后 3d分别给予二甲基纤维素和吡格列酮 (2 0mg·kg-1·d-1)连续 7d ,结肠炎症的评价包括炎症活动指数 (DAI)、大体形态损伤、组织学改变及肠黏膜髓过氧化物酶 (MPO)活性 ,采用ELISA法检测白细胞介素 (IL) 4和IL 5。结果　正常小鼠和结肠炎小鼠结肠组织LPMC凋亡率、Fas、FasL表达分别为 12 .89± 1.2 3、70 .6 3± 6 .2 4、8.5 9± 5 .4 7和 4 .2 5± 0 .84、6 2 .6 0±5 .85、2 3.75± 10 .2 3;经吡格列酮处理后 ,分别为 4 0 .5 8± 10 .32、83.98± 11.38、10 .0 4± 5 .2 1。对照组和实验组大体形态、组织学损伤、MPO值、IL 4、IL 5分别为 2 .5 0± 0 .5 5、8.83± 0 .75、3.81± 0 .17、2 16 .4 6± 34.32、10 2 .2 8± 2 5 .74和 0 .33± 0 .5 2、4 .0 0± 0 .6 3、1.2 5± 0 .16、1     

核因子-κB和激活蛋白-1在恶唑酮诱导的小鼠实验性肠炎中的表达

目的 研究恶唑酮诱导的小鼠实验性肠炎中核因子(NF)-κB和激活蛋白(AP)-1基因表达量的变化,以探索其在发病机制中的意义.方法 7～8周龄的健康昆明小鼠24只,体重25～30 g,均分为正常组和模型组.模型组小鼠采用3%恶唑酮皮肤致敏5 d后,以0.5%恶唑酮(溶解于50%乙醇中)0.15 ml一次性灌肠造模方式建立实验性结肠炎模型.3 d后处死所有小鼠,分别提取外周血单个核细胞(PBMC)、脾脏单个核细胞(SMC)和肠黏膜固有层单个核细胞(LPMC).经细胞mRNA抽提、逆转录cDNA后进行荧光定量PCR(Taqman探针法)检测NF-κB、AP-1的表达量.并进行结肠炎的组织学评分.结果 模型组NF-κB在SMC、LPMC和PBMC中的表达量均比正常组明显增高(分别为5.62±0.78比3.16±0.59、5.46±0.38比3.18±0.58、5.65±0.56比3.36±0.59,P＜0.01),AP-1亦是如此(分别为5.61±0.54比3.22±0.50、5.50±0.69比3.19±0.40、5.67±0.44比3.27±0.41,P＜0.01).结论 NF-κB、AP-1的活化可能与结肠炎的发病机制有关.     

三硝基苯磺酸和噁唑酮诱导肠炎小鼠脾脏中Th1/Th2类细胞因子基因表达的动态研究

目的 对比研究三硝基苯磺酸（TNBS）和 唑酮（OXZ）诱导的实验性小鼠肠炎模型脾脏中Th1／Th2类细胞因子基因表达的动态变化。方法 雌性BALB／c小鼠，随机分为50％乙醇对照组，OXZ诱导肠炎模型组，TNBS诱导肠炎模型组（保证每组小鼠取材时6只以上）。采用RT—PCR方法观察OXZ和TNBS诱导肠炎后小鼠脾脏于3及7d IFN-γ、IL-4 mRNA的表达变化。结果 TNBS模型组小鼠在造模后3及7d脾脏中IFN-γ mRNA的表达量均高于OXZ模型组（P〈0．05），IL-4 mRNA的表达量低于OXZ模型组（P〈0．05）。结论 在造模后3和7d TNBS模型小鼠的外周免疫皆以Th1型为主，OXZ模型小鼠的外周免疫以Th2型为主，可代表肠道局部免疫类型。     

吸烟与尼古丁对DSS肠炎小鼠免疫反应的影响机制

炎症性肠病(IBD)包括溃疡性结肠炎(UC)和克罗恩病(CD),IBD的发病机制至今不明,但是免疫功能异常在IBD发病机制中的重要作用是不容置疑的。临床流行病学研究已经证明,吸烟可以缓解UC患者的临床症状,但对CD却有恶化作用。临床观察亦发现,尼古丁灌肠和尼古丁皮贴剂对活动性UC具有治疗作用,基础研究及动物实验结果亦提示,吸烟对UC的治疗作用可能与尼古丁有关。明确吸烟/尼古丁对UC/CD的作用机制和调控环节(因素),有助于进一步澄清UC/CD在发病机制以及治疗原则等方面的异同,为IBD的预防、综合治疗提供依据。 1 材料和方法 1.1 材料野生型(购自SLC Co.,LTD.Shizuoka,Japan)和IFN-γ基因敲除(Bar Harbor,ME,USA)雄性BALB/c小鼠,6～8周龄,体质量20g～25g。饲养于日本弘前大学医学部动物实验中心,IBD模型制作依照Okayasu et al的方法,小鼠给     

吡格列酮对DSS诱导的炎症性肠病小鼠血清TNF-α与hs-CRP水平的影响

目的观察吡格列酮对炎症性肠病小鼠血清肿瘤坏死因子-α(TNF-α)与高敏C反应蛋白(hs-CRP)水平的影响,探讨吡格列酮可能成为治疗炎症性肠病的药物及降低远期动脉粥样硬化的发病风险。方法清洁BALB/c小鼠随机分为3组,分别为正常组、模型组、吡格列酮药物干预组。采用3%DSS溶液制备炎症性肠病小鼠模型,在造模第2天开始给予吡格列酮25 mg/kg灌胃,每天一次,直至实验结束。治疗结束后,采用酶联免疫法(ELISA法)检测血清hs-CRP、TNF-α。结果与正常组相比较,模型组小鼠肠组织出现炎症性表现,且血清hs-CRP、TNF-α的水平均显著增高(P0.01);与模型组相比较,吡格列酮干预组小鼠肠道炎症表现明显减轻,且血清hs-CRP、TNF-α的水平均显著降低(P0.01)。结论吡格列酮能下调炎症性肠病小鼠TNF-α的表达,减轻肠道炎症性改变,并改善肠道组织形态结构,而发挥治疗,明显降低血清hs-CRP的水平,降低远期动脉粥样硬化的风险。     

自噬激动剂雷帕霉素对实验性小鼠结肠炎的影响及作用机制初探

目的 初步探讨自噬激动剂雷帕霉素(Rapamycin,RAPA)对实验性小鼠结肠炎的影响及可能的作用机制.方法 雄性C57BL/6小鼠(6～8周)分为Control组(n=10)、DSS组(n=8)、DSS+RAPA组(n=8)、RAPA组(n=7).DSS组给予葡聚糖硫酸钠(dextran sodium sulfat...     

抗生素清肠对丁酸梭菌保护肠炎小鼠肠黏膜屏障的影响及初步机制研究

目的 评价丁酸梭菌对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的肠炎小鼠肠黏膜屏障作用和可能机制,以及抗生素清肠预处理对其的影响.方法 通过3％DSS诱导肠炎模型,并利用丁酸梭菌干预有或无抗生素清肠预处理后的小鼠,观察小鼠体质量、疾病活动度,检测小鼠病理、促炎因子TNF-α、IL-1β表达...     

吡喹酮对血吸虫肝纤维化小鼠肝脏TGF-β1和胶原表达的影响

目的 研究吡喹酮对血吸虫病肝纤维化小鼠肝脏TGF—β1和Ⅰ、Ⅲ型胶原表达的影响及其抗纤维化机制。方法 制备血吸虫病肝纤维化小鼠模型，应用免疫组化染色方法和多媒体彩色病理图文分析系统观察吡喹酮治疗前后肝脏TGF—β1和Ⅰ、Ⅲ型胶原定量表达的变化。结果 吡喹酮治疗组与感染组比较，TGF—β1，Ⅰ、Ⅲ型胶原含量明显降低(P＜0．01)。结论 吡喹酮治疗血吸虫病肝纤维化小鼠可降低其肝脏TGF—β1和Ⅰ、Ⅲ型胶原的含量而发挥抗肝纤维化作用。     

己酮可可碱对小鼠TNBS结肠炎的药效学研究

目的观察己酮可可碱(PTX)对小鼠TNBS(三硝基苯磺酸)肠炎的作用.方法通过直肠给予雄性BALB/c小鼠TNBS诱导结肠炎,应用PIX对其进行治疗,6日后收集结肠标本评价结肠炎症程度.结果直肠内给予BALB/c小鼠TNBS后可造成小鼠结肠炎性改变:FTX治疗可使小鼠疾病活动指数、结肠重量和炎症指数均显著下降(P＜0.05).结论PTX治疗可使TNBS肠炎模型小鼠肠道炎症减轻.     

转化生长因子β受体在恶唑酮结肠炎中的表达及临床意义

目的探讨转化生长因子(TGF)β信号传导通路异常在结肠炎发病中可能的作用机制。方法第0、1天,Balb/c小鼠皮肤涂搽0.2ml3%的恶唑酮,第7天以0.15ml1%的恶唑酮灌肠,第10天处死小鼠,取制模成功的6只小鼠(实验组)及6只未经处理的小鼠(对照组)的结肠组织分别采用免疫组化和蛋白免疫印迹法定性和定量检测TGFβ受体(TGFβR)Ⅰ、TGFβRⅡ、Smad7(TGFβ信号传导的抑制性介质)的表达。结果实验组小鼠均于灌肠后24h内出现厌食,懒动,稀便,便血或大便隐血(+),体重减轻。TGFβRⅠ、TGFβRⅡ、Smad7在对照组主要都表达于结肠腺体上部,在实验组都表达于整个结肠腺体及固有层、黏膜下层。TGFβRⅠ、TGFβRⅡ、Smad7在对照组和实验组的表达量分别为3.40±1.25、21.71±6.97、8.95±2.12和6.49±2.18、4.40±3.34、17.92±6.80。结论恶唑酮结肠炎中TGFβRⅠ表达无改变,TGFβRⅡ表达减少,Smad7表达增高,故该结肠炎中存在TGFβ信号传导通路异常。该模型的病理表现及免疫学特征类似人类溃疡性结肠炎。     

Effect of cyclosporine in a murine model of experimental colitis

The use of immunosuppressive therapy may be associated with significant toxicity. The aim of this study was to investigate the effect of cyclosporine A (CsA) in murine model of experimental colitis. Experimental colitis was induced in NMRI mice using an enema of 0.2% solution of dinitrofluorobenzene, combined with skin sensitization. After inducing colitis, experimental groups of animals were treated with CsA (1, 3, 5, 10, 25, 50 mg/kg/day) intraperitoneally (i.p.) or intracolonically (i.c.), and control groups were treated with phosphate-buffered saline intraperitoneally or intracolonically, respectively. Colonic inflammatory changes were assessed using a histopathologic score of 0–30, and pooled whole blood samples were processed with monoclonal antibodies for cyclosporine concentration. In addition, two groups of animals with experimental colitis were treated intraperitoneally or intracolonically with 3 mg/kg/day of CsA, and the colons were also taken for immunohistochemistry for CD25. CsA diminished the extent of colitis in groups treated with 3, 5, 10, or 25 mg/kg intraperitoneally or intracolonically, and in groups treated with 1 and 50 mg/kg intracolonically (P < 0.05). The effect of intracolonic application of CsA was not related to whole blood cyclosporine concentrations. In addition, the effect of CsA at 3 mg/kg, applied intraperitoneally or intracolonically was, in part, expressed in decreasing the numbers of CD25+ cells within colonic mucosa/submucosa (P < 0.05). In conclusions, the results of this study indicate the possibility of intracolonic application of cyclosporine in order to widen the therapeutic window for effective, but possibly toxic drug, such as cyclosporine.     

阿泰宁对噁唑酮诱导的大鼠结肠炎模型的治疗作用

目的:建立嗯唑酮诱导的大鼠结肠炎模型.观察阿泰宁对噁唑酮诱导的大鼠结肠炎的治疗作用及机制.方法:经直肠注入嗯唑酮建立大鼠溃疡性结肠炎模型.将大鼠随机分为:空白对照组(正常组,n=8),模型组(n=10),美沙拉秦组(正常组,n=10),阿泰宁组(n=10),治疗21d后处死动物,肉眼观察结肠病变,分别测体质量、结肠湿质量、脾脏质量和结肠组织病理学变化,ELISA法测定大鼠血清IL-1β、IL-10、TNF-α以及结肠黏液内容物sIgA含量,考马斯亮兰法测定血清总蛋白,溴甲酚绿比色法测定白蛋白以及肠道菌群培养.结果:模型组、阿泰宁组和美沙拉秦组大鼠体质量均比正常组显著降低(均P＜0.01).美沙拉秦组和阿泰宁组血清球蛋白含量、结肠湿质量指数较模型组差异显著(29.9±5.7,29.1±5.4vs23.7±9.5:6.0±0.9.6.2±0.4vs7.4±1.6,均P<0.05).模型组大鼠血清IL-1β和TNF-α含量较正常组、美沙拉秦组和阿泰宁组差异显著(44.6±17.2vs8.8±7.9,14.5±4.7,8.6±3.4,均P<0.01;33.5±7.2 vs 22.6±6.7,22.3±9.2,24.4±10.8,均P＜0.05).模型组大鼠血清IL-10含量较正常组、阿泰宁组差异显著(101.5±35.8vs280.5±36.1,271.3±33.8,P<0.01).阿泰宁组脾脏指数、sIgA含量比正常组显著升高(3.4±0.8vs2.7±0.3;46.0±20.3vs23.4±18.5,均P<0.05),而美沙拉秦组差异不显著.较模型组双歧杆菌数量,阿泰宁治疗后明显上升,梭杆菌数量明显下降(9.7±0.1vs9.3±0.2:3.7±0.3vs5.8±0.7,均P<0.01).结论:阿泰宁能够有效治疗噁唑酮诱导的大鼠结肠炎.     

Oxazolone‐induced murine model of ulcerative colitis

OBJECTIVE: Animal models are useful for studying disease, but there is a shortage of suitable models of ulcerative colitis. The aim of the present study was to set up an oxazolone‐induced murine colitis model and use it to research the pathogenesis of inflammatory bowel disease. METHODS: BALB/c mice were presensitized by painting the skin with 0.2 mL 3% oxazolone in 100% ethanol on days 0 and 1 followed by intrarectal administration of 0.15 mL 1% oxazolone in 50% ethanol on day 7. The disease activity index (DAI), histological changes of the colon, myeloperoxidase (MPO) activity and production of cytokines (TNF‐α, IL‐4, IFN‐γ) by the mucosa were evaluated. RESULTS: There were obvious changes in the DAI, histology and MPO activity, and the production of interleukin‐4 was markedly increased compared with the concentrations of TNF‐α and IFN‐γ, which remained normal, in the lesions. CONCLUSION: Oxazolone colitis is Th2‐mediated and has similar histologic features and distribution of inflammation to ulcerative colitis (UC), which has important implications for the use of this model in the study of the pathogenesis and treatment of UC.     

Effect of adiponectin on murine colitis induced by dextran sulfate sodium

BACKGROUND & AIMS: Adiponectin, an adipose tissue-derived hormone, exhibits anti-inflammatory properties and has various biological functions, such as increasing insulin sensitivity, reducing hypertension, and suppressing atherosclerosis, liver fibrosis, and tumor growth. The aim of the present study was to determine the effect of adiponectin on intestinal inflammation. METHODS: We investigated the effect of adiponectin on dextran sulfate sodium (DSS)-induced colitis by using adiponectin-knockout (APN-KO) mice and an adenovirus-mediated adiponectin expression system. We also examined the contribution of adiponectin deficiency to trinitrobenzene sulfonic acid (TNBS)-induced colitis. In vitro, we examined the effect of adiponectin on intestinal epithelial cells. RESULTS: After administration of 0.5% DSS for 15 days, APN-KO mice developed much more severe colitis compared with wild-type mice. The messenger RNA expression levels of chemokines were significantly higher in the colonic tissues of DSS-treated APN-KO mice compared with wild-type mice, accompanied by increased cellular infiltration, including macrophages. Adenovirus-mediated supplementation of adiponectin significantly attenuated the severity of colitis, but there were no differences in the severity of TNBS-induced colitis between the 2 groups. Adiponectin receptors were expressed in intestinal epithelial cells, and adiponectin inhibited lipopolysaccharide-induced interleukin-8 production in intestinal epithelial cells. CONCLUSIONS: Adiponectin is protective against DSS-induced murine colitis, probably due to the inhibition of chemokine production in intestinal epithelial cells and the following inflammatory responses, including infiltration of macrophages and release of proinflammatory cytokines.