联用乳酸杆菌和丁酸梭菌对小鼠急性溃疡性结肠炎的影响

目的观察联用乳酸杆菌和丁酸梭菌对小鼠急性溃疡性结肠炎的疗效并探讨其治疗机制。方法建立DSS诱导的小鼠急性溃疡性结肠炎（UC）模型,观察给予乳酸杆菌和丁酸梭菌治疗后,小鼠结肠黏膜的病理改变和TNF—α和组织因子（TF）表达的变化。结果乳酸杆菌和丁酸梭菌可明显减轻小鼠结肠黏膜的损伤;可明显抑制TNF—α和TF的表达,尤以两菌合用组抑制作用最强。结论乳酸杆菌和丁酸梭菌对DSS诱导的UC有治疗作用,对TNF-α和TF表达的协同抑制作用可能足其发挥协调治疗作用的分子机制。     

益生菌CMS-H002和CMS-H003联用对小鼠急性溃疡性结肠炎的影响

目的：研究肠道益生菌CMS-H002和CMS-H003对小鼠溃疡性结肠炎（UC）的治疗作用。方法：建立硫酸葡聚糖胺（DSS）诱导的小鼠急性UC模型。观察益生菌CMS-H002和CMS-H003治疗后，疾病活动指数（DAI）、肠道菌群及丙二醛（MDA）的变化。结果：肠道益生菌CMS-H002和CMS-H003均可明显降低DAJ的水平及结肠组织的MDA水平。疗效均优于或等于巴柳氮阳性治疗药物。治疗后，各益生菌治疗组粪便乳酸杆菌、双歧杆菌菌数均有不同程度增加；而大肠杆菌及金黄色葡萄球菌菌数均有不同程度减少；其中用CMS-H003和合用组治疗后，丁酸梭菌菌数也有增加。以合用组小鼠粪便菌群更趋近于正常小鼠粪便菌群构成。而巴柳氮阳性治疗组对小鼠粪便的菌群菌数无明显影响。结论：口服肠道益生菌CMS-H002和CMS-H003可明显改善5％DSS诱导的Balb／c小鼠急性UC的一般表现及组织学损伤。两茼舍用效果尤为明显。减少自由基的产生及逆转病态菌群是它们发挥治疗UC作用的部分机制。     

川芎嗪对溃疡性结肠炎肠粘膜过氧化物酶体增殖物激活受体γ及核因子κB的影响

目的：通过观测经川芎嗪治疗前后过氧化物酶体增殖物激活受体γ（PPARγ）、核因子κB（NF-κB）和肿瘤坏死因子（TNF）-α的变化，探讨川芎嗪治疗溃疡性结肠炎（UC）的作用机制。方法：以噁唑酮诱导小鼠UC模型，并随机分为实验对照组（CN）、模型组（OXZ）、川芎嗪治疗组（TMP）和0.9%氯化钠治疗对照组（NS）。观察UC炎症评价指标（DAI，大体、组织学损伤评分，MPO值）；采用荧光定量PCR法检测各组肠粘膜PPARγ、NF-κBp65、TNF-αmRNA的表达量；免疫组化法检测PPARγ、NF-κBp65蛋白的表达。结果：与CN组比较，OXZ组的炎症评价指标均明显增高（P〈0.01），PPARγ表达量明显下降（P〈0.01）、NF-κBp65及TNF-α表达量均显著升高（P〈0.01）。应用川芎嗪后，UC的炎症评价指标均较NS组明显降低（P〈0.01），PPARγ表达量明显升高（P〈0.01）、NF-κBp6及TNF-α表达量均下降（P〈0.01）。结论：川芎嗪对UC的结肠粘膜有保护作用，其机制可能与PPARγ的激活，NF-κB的抑制，TNF-α等炎症因子表达的减低有关。     

垂体肿瘤转化基因1通过调控肠上皮细胞焦亡在结肠炎症中的作用

目的 明确垂体肿瘤转化基因1(PTTG1)通过调控肠上皮细胞的焦亡水平在结肠炎症中的作用机制。方法 PTTG1野生型（WT）小鼠和PTTG1基因敲除（KO）小鼠各10只,随机分为4组,每组5只,分别为PTTG1 WT对照组（control组）和实验组（3%DSS组）,PTTG1 KO对照组和实验组。实验组小鼠自由饮用3%葡聚糖硫酸钠（DSS）6 d诱导急性实验性结肠炎,对照组给予无菌双蒸水。观察并记录各组小鼠的疾病活动指数（DAI）。收集小鼠结肠组织,用免疫组织化学和蛋白免疫印迹法检测焦亡相关蛋白NLRP3、ASC、GSDMD的表达水平。在人源结肠上皮细胞系（HcoEpic）中,通过shRNA沉默PTTG1的表达,TNF-α刺激细胞以诱导细胞炎症模型,检测GSDMD的表达水平。结果 DSS诱导的结肠炎小鼠结肠黏膜组织中PTTG1表达减少（P <0.01）,PTTG1敲除加重小鼠结肠炎症,PTTG1 KO实验组小鼠的结肠黏膜上皮焦亡相关蛋白表达水平上调（P <0.05）。在HcoEpic中沉默PTTG1的表达,TNF-α刺激后,细胞的GSDMD蛋白表达水平上调（P <0...     

重组硫酸软骨素蛋白SRPX2减轻小鼠葡聚糖硫酸钠诱导的结肠炎的作用

目的评价sushi重复蛋白X连锁2蛋白(SRPX2)在葡聚糖硫酸钠(DSS)诱导的实验性结肠炎小鼠中的作用及其可能机制。方法 DSS+SRPX2组,DSS组小鼠各5只,自第1天起给予5%DSS溶液自由饮用,分别给予SRPX2重组蛋白及0.9%氯化钠溶液腹腔注射5 d,至第6天处死小鼠。另设正常对照组(不予处理)。记录各组小鼠疾病活动度(DAI)评分、结肠组织学评分;检测结肠组织髓过氧化物酶(MPO)活性;分离固有层单个核细胞(LPMC),酶联免疫吸附实验法检测LPMC中肿瘤坏死因子α(TNF-α)、核因子κB(NF-κB)的表达;比色法试剂盒检测N6-腺苷酸甲基化(m~6A)修饰的RNA的水平。结果 DSS组小鼠第5天的DAI评分较正常对照组明显升高(P <0.01),DSS+SRPX2组DAI评分较DSS组均明显降低(P <0.05)。处死小鼠后,DSS组小鼠结肠组织学评分及MPO活性较正常对照组均明显升高(P <0.05),DSS+SRPX2组小鼠结肠的组织学评分及MPO活性均较DSS组明显降低(P <0.05)。DSS组小鼠LPMC中TNF-α、NF-κB水平较正常对照组均明显升高(P <0.05),DSS+SRPX2组小鼠LPMC中TNF-α、NF-κB水平均较DSS组明显降低(P <0.05)。DSS组小鼠LPMC中m~6A修饰的RNA的比例较正常对照组明显升高(P <0.05),而DSS+SRPX2组小鼠LPMC中m~6A修饰的RNA的比例较DSS组明显降低(P <0.05)。结论 SRPX2重组蛋白具有减轻DSS诱导的小鼠实验性结肠炎的作用,其作用可能与改变结肠组织LPMC中m~6A甲基化RNA的水平有关。     

乳酸杆菌和双歧杆菌粗制代谢产物对小鼠溃疡性结肠炎的影响

【目的】了解乳酸杆菌和双歧杆菌粗制代谢产物对小鼠溃疡性结肠炎(UC)的防治作用。【方法】采用右旋葡聚糖硫酸钠(DSS)建立小鼠UC模型,分别予乳酸杆菌粗制代谢产物和双歧杆菌粗制代谢产物灌肠,生理盐水(NS)和柳氮磺胺吡啶(SASP)灌肠对照,并设立空白对照组和DSS模型组对照,计算小鼠死亡率、疾病活动指数(DAI)、组织学损伤评分和组织学病理改变以及结肠黏膜IL-10免疫组化情况。【结果】①乳杆组和双歧组小鼠结肠黏膜组织病理学改变轻微,其死亡率、DAI评分、组织学损伤评分均低,与DSS模型组和NS组对照差异均有统计学意义(P<0.05)。②乳杆组和双歧组IL-10免疫组化表达均较高,与空白对照组、DSS模型组和NS组对照差异均有统计学意义(P<0.05)。【结论】①乳酸杆菌和双歧杆菌粗制代谢产物均对小鼠溃疡性结肠炎起主要的防治作用。②乳酸杆菌和双歧杆菌粗制代谢产物的某些成分通过促进IL-10表达而发挥防治作用的。     

TNFα-在小鼠叶酸诱导肾病中的致病作用及其机理研究

目的研究TNF-α在小鼠叶酸诱导肾病中的致病作用及其机制。方法建立小鼠叶酸诱导肾病模型，运用抗TNF-α多克隆抗体及对照抗体干预治疗，检测小鼠血清以及肾组织中TNF-α水平，肾小管上皮细胞凋亡情况以及凋亡相关蛋白的表达水平。结果在CD1小鼠叶酸诱导肾病模型中，尿素氮水平显著升高，肾小管上皮细胞坏死和凋亡增加，小鼠血清以及肾组织中TNF-α水平显著升高，肾皮质组织中Bcl-xL表达水平显著下降，运用抗TNF-α多克隆抗体干预治疗可保持Bcl—xL在肾组织中的表达水平，减少肾小管上皮细胞凋亡，有效减轻小鼠肾功能损害。结论TNF-α在小鼠叶酸诱导肾病发病机制中具有重要作用，阻断TNF-α是治疗急性肾衰的一种潜在有效手段。     

TNFα-在小鼠叶酸诱导肾病中的致病作用及其机理研究

目的研究TNF-α在小鼠叶酸诱导肾病中的致病作用及其机制。方法建立小鼠叶酸诱导肾病模型,运用抗TNF-α多克隆抗体及对照抗体干预治疗,检测小鼠血清以及肾组织中TNF-α水平,肾小管上皮细胞凋亡情况以及凋亡相关蛋白的表达水平。结果在CD1小鼠叶酸诱导肾病模型中,尿素氮水平显著升高,肾小管上皮细胞坏死和凋亡增加,小鼠血清以及肾组织中TNF-α水平显著升高,肾皮质组织中Bcl-xL表达水平显著下降,运用抗TNF-α多克隆抗体干预治疗可保持Bcl-xL在肾组织中的表达水平,减少肾小管上皮细胞凋亡,有效减轻小鼠肾功能损害。结论TNF-α在小鼠叶酸诱导肾病发病机制中具有重要作用,阻断TNF-α是治疗急性肾衰的一种潜在有效手段。     

针药结合对大鼠溃疡性结肠炎TNF-α、ＩＣＡＭ-1 基因表达的影响

目的:探讨针药结合对大鼠溃疡性结肠炎TNF-α、ICAM-1mRNA表达的影响。方法:采用免疫学方法制备大鼠溃疡性结肠炎模型,观察针药结合犤电针足三里和上巨虚+西药柳氮磺胺吡啶(SASP)灌胃犦、电针足三里和上巨虚及西药SASP灌胃治疗对UC大鼠脾脏和结肠粘膜TNF-α、ICAM-1 mRNA表达的影响。结果:模型组大鼠脾脏和结肠粘膜TNF-α、ICAM-1mRNA表达显著高于正常对照组;而针药结合组、电针组、西药组均有不同程度的降低,且以针药结合组最为显著。结论:针药结合可能通过调整UC大鼠TNF-α、ICAM-1的基因表达,纠正其异常的免疫功能而达到治疗目的。     

速激肽受体2对小鼠溃疡性结肠炎的影响

目的: 利用速激肽受体2（tachykinin receptor 2,Tacr2）基因敲除小鼠及诱导溃疡性结肠炎（ulcerative colitis,UC）小鼠模型,探讨Tacr2在小鼠UC发生、发展中的作用。方法: 小鼠按照基因型分成野生型组和基因敲除（纯合子）组,再按照给药与否进一步分成野生型造模组、纯合子造模组、野生型空白组、纯合子空白组。给造模组小鼠口服右旋葡聚糖硫酸钠（dextran sodium sulfate, DSS）构建UC小鼠模型,观察其UC活动指数及结肠组织中病理学改变、炎症因子及上游转录因子核因子NF-κB（nuclear factor-kappa B,NF-κB）的变化。结果: Tacr2基因敲除的纯合子小鼠经DSS诱导的UC症状比野生型造模组小鼠明显加重。进一步分析基础状态下结肠的免疫活性,发现在基础状态下,纯合子小鼠与野生型小鼠相比,结肠免疫活性明显降低,表现为结肠黏膜中 IL-1β、TNF-α、IL-6和NF-κB水平降低。结论: Tacr2对UC的发生、发展有抑制作用。     

柳氮磺胺吡啶联合酪酸梭菌活菌片治疗溃疡性结肠炎的疗效观察

目的观察联合应用柳氮磺胺吡啶和酪酸梭菌活菌片（米雅片）治疗轻、中度溃疡性结肠炎的临床疗效。方法选择41例轻、中度溃疡性结肠炎患者,随机分为治疗组22例和对照组19例。治疗组口服柳氮磺胺吡啶联合酪酸梭菌活菌片,对照组单纯口服柳氮磺胺吡啶。结果治疗组总有效率（90.91%）明显高于对照组（63.16%）（P〈0.05）。结论联合应用柳氮磺胺吡啶和酪酸梭菌活菌片比单独应用柳氮磺胺吡啶治疗轻、中度溃疡性结炎具有更好的临床疗效,能更好地改善患者的临床症状。     

Anti-inflammatory effect of taurocholate on TNBS-induced ulcerative colitis in mice

Taurocholate is a natural conjugated bile acid. The aim of this study was to evaluate the anti-inflammatory effect of taurocholate in TNBS-induced ulcerative colitis in mice. The colitis were induced by rectal administration of TNBS. After 24 h, the experimental animals were treated with sulfasalazine (SASP, 500 mg/kg/day) and taurocholate (20, 40 and 60 mg/kg) for 7 consecutive days. The anti-inflammatory effects of taurocholate for colitis were assessed by body weight, colonic weight and length, macroscopic scores, and histopathological examinations. In addition, the colonic tissue levels of myeloperoxidase (MPO) activity, interleukin (IL)-1β, interferon (IFN-γ) and tumour necrosis factor-α (TNF-α) were also determined to assess the effect of taurocholate. Compared with the model group, treatment with taurocholate (20, 40 and 60 mg/kg) significantly inhibited the body weight loss, improved colonic weight and length, and decreased macroscopic and histopathological scores. Furthermore, the activity accumulation of MPO and the colonic tissue levels of IL-1β, IFN-γ and TNF-α were also decreased by administration of taurocholate. All the findings of this study suggested that taurocholate has the anti-inflammatory effect in ulcerative colitis in mice and indicated it as a good candidate to treat inflammatory bowel disease.     

Blockade of CXCL12/CXCR4 axis ameliorates murine experimental colitis

Recent studies indicate that the CXCL12/CXCR4 interaction is involved in several inflammatory conditions. However, it is unclear whether this interaction has a role in the pathophysiology of inflammatory bowel disease (IBD). We investigated the significance of this interaction in patients with IBD and in mice with dextran sulfate sodium (DSS)-induced colitis and the effect of a CXCR4 antagonist on experimental colitis. First, we measured CXCR4 expression on peripheral T cells in patients with IBD. Furthermore, we investigated CXCR4 expression on leukocytes and CXCL12 expression in the colonic tissue of mice with DSS-induced colitis, and we evaluated the effects of a CXCR4 antagonist on DSS-induced colitis and colonic inflammation of interleukin (IL)-10 knockout (KO) mice. Colonic inflammation was assessed both clinically and histologically. Cytokine production from mesenteric lymph node cells was also examined. CXCR4 expression on peripheral T cells was significantly higher in patients with active ulcerative colitis (UC) compared with normal controls, and CXCR4 expression levels of UC patients correlated with disease activity. Both CXCR4 expression on leukocytes and CXCL12 expression in colonic tissue were significantly increased in mice with DSS-induced colitis. Administration of a CXCR4 antagonist ameliorated colonic inflammation in DSS-induced colitis and IL-10 KO mice. CXCR4 antagonist reduced tumor necrosis factor-alpha and interferon-gamma production from mesenteric lymph node cells, whereas it did not affect IL-10 production. The percentage of mesenteric Foxp3+CD25+ T cells in DSS-induced colitis was not affected by CXCR4 antagonist. These results suggest that blockade of this chemokine axis might have potential as a therapeutic target for the treatment of IBD.     

Glucagon-like peptide-2 and common therapeutics in a murine model of ulcerative colitis

The intestinal hormone glucagon-like peptide-2 (GLP-2) enhances bowel growth and reduces the severity of colonic injury in dextran sulfate sodium (DSS)-induced colitis in mice. In humans, ulcerative colitis is normally treated with aminosalicylates (ASAs) and corticosteroids (CSs) to reduce inflammation. However, whether the intestinotropic effects of GLP-2 are altered when combined with ASAs and/or CSs has not previously been explored. Thus, each agent [vehicle, ASA (sulfasalazine), CS (methylprednisolone), and ASA + CS] was administered alone or with GLP-2 to normal mice or mice with 3.5% DSS in the drinking water, for 10 consecutive days. GLP-2 treatment of DSS-mice increased survival and small intestinal weight (p < 0.05), and decreased body weight loss and colonic damage (p < 0.05). Furthermore, GLP-2 increased the number of proliferating cells in the colonic crypts of DSS-mice (p < 0.05). Administration of ASA, CS, or ASA + CS alone did not affect growth of the intestine in DSS-mice. However, administration of GLP-2 in combination with ASA was permissive for the beneficial effects of GLP-2 on survival and colonic damage, whereas CS treatment prevented these effects of GLP-2. Concomitant administration of GLP-2 with ASA + CS resulted in intermediate effects. No differences between colonic myeloperoxidase activity or IkappaB levels (an inhibitor of the nuclear factor-kappaB pro-inflammatory pathway) were found for any of these therapeutic agents. When taken together, the ability of GLP-2 to protect colonic mucosal architecture in DSS-colitis, and its effectiveness when given in combination with ASA, but not with CS, suggests a novel approach for the treatment of patients with colitis.     

Hepatocyte growth factor facilitates colonic mucosal repair in experimental ulcerative colitis in rats

Hepatocyte growth factor (HGF) modulates intestinal epithelial cell proliferation and migration, serving as a critical regulator of intestinal wound healing. In this study, we examined the effect of administration of recombinant human HGF on colonic mucosal damage in vivo. Acute colitis was induced in rats by feeding with 5% dextran sulfate sodium (DSS) for 7 days, and colitis was subsequently maintained by feeding with 1% DSS. On the 5th day of DSS administration, osmotic pumps releasing recombinant human HGF (200 microg/day) were implanted into the peritoneum of the rats. Continuous intraperitoneal delivery of HGF led to both increased serum human HGF levels and c-Met tyrosine phosphorylation within the colonic mucosa. Compared with mock-treated rats, those administered human HGF showed a reduction in colitis-associated weight loss, large intestinal shortening, and improved colonic erosions. Enhanced epithelial regeneration and cellular proliferation were observed in rats treated with recombinant human HGF. The weights of the liver, kidneys, and spleen were not affected by HGF administration. These results indicate that HGF administration accelerates colonic mucosal repair in rats with DSS-induced colitis and suggest that recombinant human HGF may be a useful therapeutic tool to facilitate intestinal wound healing in patients with ulcerative colitis.     

Rebamipide suppresses TLR-TBK1 signaling pathway resulting in regulating IRF3/7 and IFN-α/β reduction

TANK-binding kinase 1 (TBK1) regulates the interferon regulatory factor (IRF) 3 and IRF7 activation pathways by double strand RNA (dsRNA) via Toll-like receptor (TLR) 3 and by lipopolysaccharide (LPS) via TLR4. Rebamipide is useful for treating inflammatory bowel disease (IBD). Although IBD is associated with nuclear factor-κB (NF-κB), any association with the TBK1-IRF pathway remains unknown. How rebamipide affects the TBK1-IRF pathway is also unclear. We analyzed the relationship between IBD (particularly ulcerative colitis; UC) and the TLR-TBK1-IRF3/7 pathway using human colon tissue, a murine model of colitis and human colonic epithelial cells. Inflamed colonic mucosa over-expressed TKB1, NAP1, IRF3, and IRF7 mRNA compared with normal mucosa. TBK1 was mainly expressed in inflammatory epithelial cells of UC patients. The expression of TBK1, IRF3, IRF7, IFN-α and IFN-β mRNA was suppressed in mice given oral dextran sulfate-sodium (DSS) and daily rectal rebamipide compared with mice given only DSS. Rebamipide reduced the expression of TBK1, IRF3 and IRF7 mRNA induced by LPS/dsRNA, but not of NF-κB mRNA in colonic epithelial cells. Rebamipide might suppress the TLR-TBK1 pathway, resulting in IRF3/7-induction of IFN-α/β and inflammatory factors. TBK1 is important in the induction of inflammation in patients with UC. If rebamipide represses the TLR-TBK1 pathway, then rectal administration should suppress inflammation of the colonic mucosa in patients with UC.