酪酸菌对大鼠右旋葡聚糖硫酸钠结肠炎的治疗作用

酪酸菌 (宫入菌 )作为肠道正常菌群之一 ,已被广泛用于治疗肠道菌群失调所致的急慢性腹泻、肠易激综合征、抗生素相关性肠炎等。另一方面 ,酪酸菌产生的酪酸是结肠上皮细胞的主要能量来源之一 ,有促进结肠上皮细胞增殖的作用[1] 。已有大量资料表明 ,用酪酸灌肠治疗溃疡性结肠炎有效 ,在此我们观察酪酸菌口服治疗大鼠右旋葡聚糖硫酸钠(DSS)结肠炎的效果。一、材料和方法1.实验动物 :4周龄SD雄性大鼠 30只。2 .结肠炎模型的诱发 :用 3%DSS(和光纯药工业会社生产 ,相对分子质量 5 0 0 0 )溶液给大鼠 ,自由饮水 ,连续 10d。3.治疗实验 :治…     

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。     

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。     

葡聚糖硫酸钠致溃疡性结肠炎大鼠模型的肠道微生态

目的:采用多聚酶链反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术研究葡聚糖硫酸钠(dextran sulfate sodium,DSS)所致溃疡性结肠炎(ulcerative colitis,UC)模型大鼠肠道菌群的多样性和丰度,从"肠道微生态"这一全新角度为该模型提供更丰富的数据,也为深入探讨UC的发生发展提供新的实验依据.方法:26只健康♂SD大鼠随机分为空白组和UC模型组,模型组饮用4%的DSS溶液7d复制UC模型.收集两组大鼠粪便排泄物,采用PCR-DGGE法研究肠道菌群的多样性、相似性及丰度;对优势条带进行回收测序,鉴定菌种.所得结果及数据采用Quantity one、Chromas、MGAE5、SIMCA-P+及SPSS18.0等软件进行分析.结果:DSS诱导UC后7d,模型大鼠出现便血、病理形态学改变等典型的UC炎性病变特征.样本优势条带菌种鉴定显示大鼠的肠道细菌主要归属于拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)及变形菌门(Proteobacteria)3大类,但与空白组相比,模型组肠道菌群的丰度及多样性指数明显降低(P<0.05).菌种鉴定表明UC组乳酸杆菌(Lactobacillus sp)和毛螺科菌(Lachnospiraceae bacterium)菌种显著较少,而双酶梭菌(Clostridium bifermentans)等菌种含量明显增多(均P<0.05).     

T辅助细胞在右旋葡聚糖硫酸钠诱导的小鼠结肠炎中的作用研究

目的　探讨Th1/Th2细胞在右旋葡聚糖硫酸钠 (DSS)诱导小鼠结肠炎中的作用。方法　利用细胞内细胞因子流式细胞检测法 ,测定DSS诱导急、慢性结肠炎小鼠肠黏膜和脾脏单核细胞 (SMC)中Th1/Th2比例。结果　①结肠炎急性期 ,小鼠肠黏膜固有层单核细胞 (LPMC)中Th1细胞为 (8.90± 1.2 3) % ,较对照组升高 (P <0 .0 5 ) ;Th2细胞占 (3.2 5± 1.2 5 ) % ,与对照组差异无显著性 (P >0 .0 5 ) ;Th1/Th2比例为 3.0 9± 1.18,较对照组升高 (P <0 .0 5 )。②在慢性期 ,肠黏膜LPMC中Th1、Th2细胞分别占 (5 .5 2± 1.2 8) %和 (10 .0 8± 1.75 ) % ,均较对照组升高 (P <0 .0 5 ) ;Th1/Th2比例为 0 .5 2± 0 .2 1,较对照组降低 (P <0 .0 5 )。脾脏SMC中Th1、Th2百分比、Th1/Th2比例 ,各期无明显差异。结论　DSS结肠炎是一种以肠黏膜免疫功能紊乱为主的炎症性病变。Th1优势反应可能与急性期损伤有关 ,而Th2优势反应可能在炎症的慢性化过程中发挥重要作用。     

肝素治疗右旋葡聚糖硫酸钠诱导小鼠结肠炎的疗效观察

目的　评价肝素预防及治疗右旋葡聚糖硫酸钠 (DSS)诱导小鼠结肠炎的有效性。方法　32只小鼠中 ,16只正常小鼠分成两组 ,饮用DSS 7d的同时预防组皮下注射肝素 ,对照组皮下注射生理盐水 ;另 16只饮用DSS 7d后诱导结肠炎的小鼠分成两组 ,治疗组皮下注射肝素 ,对照组皮下注射生理盐水。用疾病活动指数、组织学评分、TNF α的表达和马休斯猩红兰 (MSB)检测微血栓评价疗效。结果　肝素在预防组降低微血栓的形成 ,对照组 8只中 4只微血栓阳性 ,预防组均阴性 (P =0 .0 38)。治疗组组织学评分、TNF α的表达明显降低 ,治疗组与对照组的直肠、横结肠组织学评分和TNF α的表达分别为 1.33和 1.85 (P <0 .0 5 ) ,0 .92和 1.6 8(P <0 .0 5 ) ,(5 .5± 3.5 ) %和 (10 .8± 4.2 ) % (P <0 .0 5 )。结论　肝素可抑制DSS诱导结肠炎小鼠血栓形成和结肠炎症 ,实验结果提示肝素用于溃疡性结肠炎治疗也可能有效     

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。     

凝结芽孢杆菌TBC169株对亚急性衰老模型小鼠的抗衰老作用

目的 探讨凝结芽孢杆菌对亚急性衰老模型小鼠的抗衰老作用.方法 用D-半乳糖复制小鼠亚急性衰老模型,监测小鼠脑、心、肝组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活力和丙二醛(MDA)的水平并观察高、低剂量的凝结芽孢杆菌对这些指标的影响.结果 低剂量凝结芽孢杆菌能显著改善衰老模型小鼠脑组织SOD、 GSH-Px活性(P＜0.05),极显著改善肝组织GSH-Px活性、心脏组织SOD活性及降低心脏组织MDA水平(P＜0.01);高剂量凝结芽孢杆菌可显著降低肝组织MDA水平(P＜0.05)和极显著改善心脏组织GSH-Px活性(P＜0.01).结论 凝结芽孢杆菌TBC169株可诱导抗衰老酶合成,减少产生或加速清除自由基,起到延缓衰老作用.     

肥大细胞在葡聚糖硫酸钠诱导大鼠结肠炎中的致炎作用研究

目的 研究肠黏膜且巴大细胞(IMMC)在葡聚糖硫酸钠(DSS)诱导结肠炎时的致炎作用。方法 SD大鼠18只,随机分为DSS诱导大鼠结肠炎模型组(10只)和正常对照组(8只)。1周内模型组饮用3％DSS溶液,对照组饮水,观察大鼠的腹泻、便血症状及结肠组织学改变,用荧光法测定结肠组织组胺浓度,ELISA法测定结肠组织肿瘤坏死因子-a(TNF -a)和前列腺素E2(PGE2)水平.组化法对IMMC染色并记数。结果 DSS诱导的模型组第7天开始出现腹泻、便血,结肠黏膜组织学检查有明显损伤,和对照组比较:结肠组织组胺水平降低,TNF-a和PGE2显著升高,IMMC计数增多(P均<0.05),脱颗粒现象明显。结论结肠IMMC具有明显的致炎作用,其机制是IMMC活化及促组胺释放,引发肥大细胞--细胞因子级联反应,促进TNF-a和PGE2的生成,导致腹泻,便血等症状。     

葡聚糖硫酸钠诱导小鼠实验性结肠炎的临床和组织病理学特征研究

目的研究葡聚糖硫酸钠(DSS)诱导小鼠实验性结肠炎的临床和病理特征,以便于指导选择合适的溃疡性结肠炎(ulcerative colitis,UC)小鼠模型。方法予3%DSS溶液喂饲野生型C57BL/6成年小鼠诱导急性结肠炎模型,分别于第0天、第3天、第5天、第7天和第10天麻醉处死小鼠,取结肠观察不同时期结肠病理学特征,造模过程中连续观察并记录小鼠体质量、大便和死亡情况。结果小鼠造模第3天开始出现粪便潜血,第5天开始出现血便,第10天开始出现死亡小鼠,死亡率为30%。DSS诱导小鼠急性结肠炎模型疾病活动指数第3天后与饮用纯净水的小鼠比较明显增高(P0.05)。小鼠结肠炎造模第3天黏膜上皮细胞开始逐渐丧失,第7~10天最为严重;隐窝结构的紊乱从第3天开始发生,第7天出现固有层塌陷;炎性细胞浸润从第3天开始数量逐渐增加,第10天最为严重。DSS诱导小鼠急性结肠炎模型结肠组织病理评分第5天后与饮用纯净水的小鼠比较明显增高(P0.05)。结论 3%DSS诱导野生型C57BL/6成年小鼠急性结肠炎模型可用于UC的实验研究,第3天出现明显的炎症表现,第7天模型较理想,小鼠死亡少。     

Antibody to eosinophil cationic protein suppresses dextran sulfate sodium-induced colitis in rats

AIM: To produce an antibody against rat eosinophil cationic protein (ECP) and to examine the effects of the antibody in rats with dextran sulfate sodium (DSS)-induced colitis. METHODS: An antibody was raised against rat ECP. Rats were treated with 3% DSS in drinking water for 7 d and received the antibody or normal serum. The colons were examined histologically and correlated with clinical symptoms. Immunohistochemistry and Western blot analysis were estimated as a grade of inflammation. RESULTS: The ECP antibody stained the activated eosinophils around the injured crypts in the colonic mucosa. Antibody treatment reduced the severity of colonic ulceration and acute clinical symptoms (diarrhea and/or bloodstained stool). Body weight gain was significantly greater and the colon length was significantly longer in anti-ECP-treated rats than in normal serum-treated rats. Expression of ECP in activated eosinophils was associated with the presence of erosions and inflammation. The number of Ki-67-positive cells in the regenerated surface epithelium increased in anti-ECP-treated rats compared with normal serum-treated rats. Western blot analysis revealed reduced expression of macrophage migration inhibitory factor (MIF) in anti-ECP-treated rats. CONCLUSION: Our results indicate that treatment with ECP antibody, improved DSS-induced colitis in rats, possibly by increasing the regenerative activity of the colonic epithelium and downregulation of the immune response, and suggest that anti-ECP may promote intestinal wound healing in patients with ulcerative colitis (UC).     

糖分子探针用于检测溃疡性结肠炎大鼠肠道黏膜通透性的研究

目的运用糖分子探针检测葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)大鼠肠道黏膜通透性的改变。方法 24只健康SD大鼠随机分成两组:UC组和正常对照组,观察结肠黏膜病变,应用糖分子探针检测肠道黏膜通透性。对比两组检测值,评价大鼠肠道黏膜通透性的改变。结果 (1)UC组病变主要累及黏膜及黏膜下层,可见大量炎性细胞浸润,以中性粒细胞为主,部分表面上皮脱落,上皮内杯状细胞减少,隐窝破坏。(2)与正常对照组相比,UC组大鼠乳果糖及三氯蔗糖排出量明显升高,甘露醇排出量明显降低,乳果糖/甘露醇比值明显升高。结论糖分子探针可以有效的检测大鼠肠道黏膜通透性的改变,反映肠道黏膜的损伤。     

New approach of medicinal herbs and sulfasalazine mixture on ulcerative colitis induced by dextran sodium sulfate

BACKGROUND Sulfasalazine has been used as a standard-of-care in ulcerative colitis for decades, however, it results in severe adverse symptoms, such as hepatotoxicity, blood disorders, male infertility, and hypospermia. Accordingly, the new treatment strategy has to enhance pharmacological efficacy and stimultaneously minimize side effects.AIM To compare the anti-inflammatory action of sulfasalazine alone or in combination with herbal medicine for ulcerative colitis in a dextran sodium sulfate(DSS)-induced colitis mouse model.METHODS To induce ulcerative colitis, mice received 5% DSS in drinking water for 7 d. Animals were divided into five groups(n = 9 each) for use as normal(non-DSS), DSS controls, DSS + sulfasalazine(30 mg/kg)-treatment experimentals, DSS + sulfasalazine(60 mg/kg)-treatment experimentals, DSS + sulfasalazine(30 mg/kg) + Citrus unshiu peel and Bupleuri radix mixture(30 mg/kg)(SCPB)-treatment experimentals.RESULTS The SCPB treatment showed an outstanding effectiveness in counteracting the ulcerative colitis, as evidenced by reduction in body weight, improvement in crypt morphology, increase in antioxidant defenses, down-regulation of proinflammatory proteins and cytokines, and inhibition of proteins related to apoptosis.CONCLUSION SCPB may represent a promising alternative therapeutic against ulcerative colitis, without inducing adverse effects.     

瑞巴匹特预防葡聚糖硫酸酯钠诱发的小鼠结肠炎疗效观察及作用机制初探

目的近年氧自由基在溃疡性结肠炎(UC)发病过程中作用受到关注,一种新型的黏膜保护剂瑞巴匹特被认为具有清除氧自由基的作用,有望成为治疗溃疡性结肠炎的新药物。本文观察瑞巴匹特灌肠和灌胃治疗葡聚糖硫酸酯钠(DSS)诱发的小鼠结肠炎效果并探讨可能的作用机制。方法 3%DSS予8周龄雄性BALB/c小鼠自由饮用7 d制成小鼠结肠炎模型。予DSS前5 d开始瑞巴匹特(45 mg/kg/d)灌肠或灌胃治疗直到造模结束,处死小鼠取结肠组织,测量小鼠体重、结肠长度,进行大体和病理评分,分光光度法测定髓过氧化物酶、丙二醛含量,免疫组化法测定核因子κB(NFκB)表达水平,RT-PCR法测定过氧化物酶体增殖体激活受体γ(PPARγ)mRNA表达。结果与安慰剂对照组比较,瑞巴匹特灌肠和灌胃治疗组小鼠大体和病理评分显著改善,髓过氧化物酶活性、丙二醛含量、NFκB活性明显降低,PPARγmRNA的表达显著升高。结论瑞巴匹特可有效预防DSS诱发的小鼠结肠炎,其抑制炎症作用至少部分与清除氧自由基,从而维持局部PPARγ表达和抑制NFκB活性有关。     

Systems pharmacology approach reveals the antiinflammatory effects of Ampelopsis grossedentata on dextran sodium sulfate-induced colitis

AIM To investigate the protective effects of Ampelopsis grossedentata(AMP) on dextran sulfate sodium(DSS)-induced colitis in mice based on systems pharmacology approach.METHODS Systems pharmacology approach was used to predict the active ingredients, candidate targets and the efficacy of AMP on ulcerative colitis(UC) using a holistic process of active compound screening, target fishing, network construction and analysis. A DSSinduced colitis model in C57 BL/6 mice(n = 10/group) was constructed and treated with 5-aminosalicylic acid(100 mg/kg/d) and AMP(400 mg/kg/d) to confirm the underlying mechanisms and effects of AMP on UC with western blot analyses, polymerase chain reaction, histological staining and immunohistochemistry.RESULTS The therapeutic effects of AMP against DSS-induced colitis were determined in the beginning, and the results showed that AMP significantly improved the disease in general observations and histopathology analysis. Subsequent systems pharmacology predicted 89 corresponding targets for the four candidate compounds of AMP, as well as 123 candidate targets of UC, and protein-protein interaction networks were constructed for the interaction of putative targets of AMP against UC. Enrichment analyses on TNF-α and RANKL/RANK, a receptor activator of NF-κB signaling pathways, were then carried out. Experimental validation revealed that inflammation-related signaling pathways were activated in the DSS group, and AMP significantly suppressed DSS-induced high expression of IRAK1, TRAF6, IκB and NF-κB, and inhibited the elevated expression levels of TNF-α, IL-1β, IL-6 and IL-8.CONCLUSION AMP could exert protective effects on UC via suppressing the IRAK1/TRAF6/NF-κB-mediated inflammatory signaling pathways.     

Evaluation of the effect of pyrrolidine dithiocarbamate in suppressing inflammation in mice with dextran sodium sulfate-induced colitis

AIM: To evaluate the effect of pyrrolidine dithio- carbamate (PDTC; an NF-κB inhibitor) administered at low (50 mg/kg) and high (100 mg/kg) doses in suppressing colitis in mice with dextran sodium sulfate (DSS)-induced colitis. METHODS: Mice were divided into a DSS-untreated group (normal group), DSS-treated control group, DSS+PDTC-treated groupⅠ(low-dose group), and DSS+PDTC-treated groupⅡ (high-dose group). In each group, the disease activity index score (DAI score), intestinal length, histological score, and the levels of activated NF-κB and inflammatory cytokines (IL-1β and TNF-α) in tissue were measured. RESULTS: The DSS+PDTC-treated groupⅡ exhibited suppression of shortening of intestinal length and reduction of DAI score. Activated NF-κB level and IL-1β and TNF-α levels were significantly lower in DSS+PDTC- treated groupⅡ. CONCLUSION: These findings suggest that PDTC is useful for the treatment of ulcerative colitis.