同型半胱氨酸调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的实验研究

目的探讨同型半胱氨酸(homocysteine,Hcy)是否通过调控MEK-ERK-MLCK通路影响结肠炎大鼠肠黏膜通透性的机制。方法 SD大鼠分为4组,A组为正常对照组(NS皮下注射+NS灌肠),B组为正常对照+Hcy注射组(Hcy皮下注射+NS灌肠),C组为TNBS模型组(NS皮下注射+TNBS灌肠),D组为TNBS模型+Hcy注射组(Hcy皮下注射+TNBS灌肠)。建立高Hcy血症的实验性结肠炎大鼠模型,实验结束时取大鼠结肠组织病理学检查,并进行结肠匀浆检测MPO活性,采用Western blot方法检测大鼠小肠组织中MEK、ERK、p-ERK、MLCK、p-MLCK的蛋白表达水平,采用RT-q PCR方法检测大鼠小肠组织中MLCK mRNA表达。结果与正常对照组及模型对照组相比,TNBS模型+Hcy皮下注射组大鼠DAI及HI评分增高,结肠匀浆MPO活性增高,小肠黏膜组织MEK、ERK、p-ERK、MLCK、p-MLCK蛋白表达水平增加,MLCK mRNA相对表达量增加。结论 Hcy增加实验性结肠炎大鼠肠黏膜通透性,可能与调控MEK-ERKMLCK信号通路有关。     

消退素D1抑制NLRP3信号通路对DSS结肠炎小鼠的影响

目的探讨消退素D1(resolvin D1,RvD1)对实验性结肠炎小鼠肠道炎症的影响及可能机制。方法将C57BL/6小鼠分为4组,包括正常组、RvD1对照组、葡聚糖硫酸钠(dextran sodium sulfate, DSS)模型组、RvD1治疗组。实验结束后,分离小鼠小肠及结肠组织。测量疾病活动指数(DAI),病理学评分(HI),检测髓过氧化物酶(MPO)活性水平,伊文思蓝检测肠黏膜通透性,电镜检测肠上皮细胞连接及细胞因子水平。分析NLRP3炎症小体相关基因表达水平。结果与正常组小鼠相比,模型组DAI评分、HI评分、MPO活性水平,以及结肠组织匀浆中促炎细胞因子的产生明显增加(P<0.05)。RvD1组小鼠上述实验指标明显减低(P<0.05)。模型组小鼠结肠组织NLRP3炎症小体表达水平增加(P<0.05);RvD1处理1周,小鼠结肠组织NLRP3通路相关蛋白表达水平降低(P<0.05)。结论 RvD1具有改善DSS结肠炎小鼠肠道炎症的作用,其机制可能与抑制NLRP3炎性体信号通路有关。     

特异性Kv1.3阻滞剂调节巨噬细胞自噬在抗结肠炎中的作用

目的探讨特异性Kv1. 3阻滞剂PAP-1抗DSS结肠炎小鼠的作用机制。方法将C57BL/6小鼠随机分为正常对照组、PAP-1对照组、DSS模型组、DSS+PAP-1组。5%DSS制备结肠炎模型,PAP-1 (3μg·g-1,每天3次,连续7d)腹腔注射给药。进行DAI评分;实验结束行结肠HE染色评分,结肠组织匀浆检测MPO活性及炎症因子水平;免疫荧光显微镜观察结肠巨噬细胞改变;透射电镜观察巨噬细胞自噬结构; q PCR、Western blot检测小鼠结肠和巨噬细胞i NOS、IL-1β及自噬相关标志物表达。结果 PAP-1减少DSS结肠炎小鼠体质量的下降,降低DAI及结肠HI评分(P <0. 05);PAP-1降低结肠MPO活性及促炎因子水平(P <0. 05)。PAP-1降低结肠F4/80表达,使腹腔巨噬细胞及脾巨噬细胞内自噬泡增多;同时,PAP-1降低结肠、腹腔巨噬细胞和脾巨噬细胞i NOS、IL-1β及p62表达,升高LC3-Ⅱ及Beclin-1表达(P <0. 05)。结论 PAP-1可能通过减少结肠炎小鼠巨噬细胞自噬受损,降低炎症因子水平,减轻结肠炎症损害。     

枫蓼提取物对小鼠实验性溃疡性结肠炎的防治作用

目的探讨枫蓼提取物对小鼠溃疡性结肠炎的防治作用。方法通过小肠推进实验和番泻叶致小鼠腹泻模型观察小肠推进率、稀便率及腹泻指数。用4%葡聚糖硫酸钠(DSS)建立小鼠溃疡性结肠炎模型,对正常组、模型组、美沙拉嗪组、枫蓼提取物11.7,23.4和46.8 g·kg-1剂量组小鼠进行疾病活动指数(DAI)评分,测定结肠组织中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、丙二醛(MDA)和一氧化氮(NO)含量及髓过氧化物酶(MPO)的活性。结果枫蓼提取物46.8 g·kg-1能抑制小鼠小肠推进率,显著减少番泻叶致腹泻小鼠腹泻次数,降低腹泻率和腹泻指数(P<0.05)。结肠炎结果显示,与正常组比较,模型组小鼠体质量减轻,DAI评分增高(P<0.05),结肠组织中MPO活性、IL-1β、TNF-α、MDA和NO含量升高(P<0.01)。与模型组比较,枫蓼提取物46.8 g·kg-1组小鼠DAI评分降低29.1%(P<0.05);23.4和46.8 g·kg-1组结肠组织中MPO活性、TNF-α、MDA和NO含量均降低(P<0.05)。结论枫蓼提取物能改善DSS诱导小鼠溃疡性结肠炎,可能与解痉、止泻、抗炎、抗氧化及减少炎性介质释放等有关。     

丹参酮IIA通过调控中性粒细胞活性减轻DSS诱导的小鼠急性结肠炎北大核心CSCD

目的观察丹参酮IIA对葡聚糖硫酸钠(DSS)诱导的小鼠实验性结肠炎是否有干预效果以及对结肠黏膜中性粒细胞浸润和活化的影响,旨在探讨丹参酮IIA治疗结肠炎的作用机制。方法随机将30只C57BL/6小鼠分为正常对照组、DSS组和DSS+丹参酮IIA干预组,每组10只动物;其中模型组给予3%DSS溶液喂饲7 d,干预组小鼠在给予3%DSS溶液喂饲的同时给予丹参酮IIA(200 mg/kg)腹腔注射,1次/d;期间对小鼠疾病活动指数(DAI)进行评分。染毒7 d后处死小鼠,取结肠组织进行病理学检查;采用异硫氰酸荧光素标记葡聚糖(FITC-dextran)法检测小鼠肠黏膜通透性;采用免疫荧光染色方法检测结肠组织Ly6G的表达,并检测MPO活性和炎症因子的水平。结果和正常对照组相比,DSS组小鼠第7天时出现体重减轻、便血和腹泻等症状,DAI显著增高。丹参酮IIA干预明显减轻小鼠结肠炎性损伤,表现为干预组小鼠DAI低于DSS组,差异有统计学意义(P<0.05);干预组小鼠结肠组织病理损伤显著减轻,肠黏膜穿透性亦低于DSS组(P=0.032)。此外,和DSS组相比,丹参酮IIA干预组小鼠结肠组织中性粒细胞浸润和活化程度明显减少,并伴有炎症因子水平显著下降(P<0.05)。结论在本实验条件下,丹参酮IIA对小鼠实验性结肠炎有明显的治疗作用,其机制可能与抑制中性粒细胞浸润、活化和减少炎症因子生成有关。     

丹参酮IIA通过调控中性粒细胞活性减轻DSS诱导的小鼠急性结肠炎

目的观察丹参酮IIA对葡聚糖硫酸钠(DSS)诱导的小鼠实验性结肠炎是否有干预效果以及对结肠黏膜中性粒细胞浸润和活化的影响,旨在探讨丹参酮IIA治疗结肠炎的作用机制。方法随机将30只C57BL/6小鼠分为正常对照组、DSS组和DSS+丹参酮IIA干预组,每组10只动物;其中模型组给予3%DSS溶液喂饲7 d,干预组小鼠在给予3%DSS溶液喂饲的同时给予丹参酮IIA(200 mg/kg)腹腔注射,1次/d;期间对小鼠疾病活动指数(DAI)进行评分。染毒7 d后处死小鼠,取结肠组织进行病理学检查;采用异硫氰酸荧光素标记葡聚糖(FITC-dextran)法检测小鼠肠黏膜通透性;采用免疫荧光染色方法检测结肠组织Ly6G的表达,并检测MPO活性和炎症因子的水平。结果和正常对照组相比,DSS组小鼠第7天时出现体重减轻、便血和腹泻等症状,DAI显著增高。丹参酮IIA干预明显减轻小鼠结肠炎性损伤,表现为干预组小鼠DAI低于DSS组,差异有统计学意义(P0.05);干预组小鼠结肠组织病理损伤显著减轻,肠黏膜穿透性亦低于DSS组(P=0.032)。此外,和DSS组相比,丹参酮IIA干预组小鼠结肠组织中性粒细胞浸润和活化程度明显减少,并伴有炎症因子水平显著下降(P0.05)。结论在本实验条件下,丹参酮IIA对小鼠实验性结肠炎有明显的治疗作用,其机制可能与抑制中性粒细胞浸润、活化和减少炎症因子生成有关。     

布地奈德脂质体温敏凝胶对溃疡性结肠炎模型小鼠的药效研究

目的:研究布地奈德脂质体温敏凝胶剂(Bud-Lip-TSG)对葡聚糖硫酸钠(dextran sulfate sodium salt,DSS)致急性溃疡性结肠炎(ulcerative colitis,UC)昆明小鼠的药效作用。方法:建立DSS诱导的小鼠UC模型。实验动物分为正常对照组、模型组、阳性对照组及Bud-Lip-TSG低、中、高剂量组。测定小鼠给药前后疾病活动指数(DAI)、肠黏膜损伤评分(CMDI)、脏器指数、结肠组织病理评分,血清中IL-6、IL-10水平,结肠组织中的TNF-α、IL-10、髓过氧化酶(MPO)水平变化。结果:Bud-Lip-TSG各剂量组小鼠的DAI评分、CMDI、结肠组织病理评分、脾脏指数降低;结肠长度、结肠湿质量、结肠指数增加;各给药组结肠组织中IL-10显著升高,TNF-α水平、MPO活力显著降低,;Bud-Lip-TSG高剂量组血液中IL-6显著降低、IL-10显著升高,Bud-Lip-TSG中、高剂量组胸腺指数显著增加。结论:Bud-Lip-TSG可改善模型小鼠UC症状,对结肠黏膜有修复作用,可改善炎症因子的表达失衡状态。     

乳酸菌对DSS诱导的溃疡性结肠炎小鼠肠道通透性影响

目的观察乳酸菌制剂对溃疡性结肠炎(UC)小鼠肠黏膜通透性影响。方法采用葡聚糖硫酸钠(DSS)诱导小鼠UC模型。小白鼠60只分为4个组,正常对照组、模型对照组、乳酸菌1组、乳酸菌2组。乳酸菌1组、2组用菌液灌胃,正常对照组模型对照用0.9%氯化钠溶液灌胃。9 d后各组小鼠每日记录疾病活动指数(DAI)评分,观察小鼠结肠黏膜组织学改变,应用酶联免疫法检测小鼠血D-乳酸含量。结果与模型组比较,乳酸菌1组、2组小鼠DAI与组织学损伤评分(CMDI)均下降(P<0.01),血D-乳酸的含量明显降低(P<0.01),乳酸菌1组、2组各指标差异无统计学意义。结论乳酸菌制剂灌胃可降低溃疡性结肠炎小鼠结肠黏膜通透性。     

具备高同型半胱氨酸血症的实验性结肠炎模型的建立

目的建立具备高同型半胱氨酸血症(HHcy)的实验性结肠炎模型,初步探讨Hcy对结肠炎症损伤的影响。方法将SD大鼠分为4组:正常对照组(NS灌肠+皮下注射NS)、正常对照+Hcy注射组(NS灌肠+皮下注射Hcy)、TN-BS模型对照组(TNBS/乙醇灌肠+皮下注射NS)、TNBS模型+Hcy注射组(TNBS/乙醇灌肠+皮下注射Hcy)。以TN-BS/乙醇混合溶液(100 mg·kg-1TNBS加入等体积NS)灌肠1次制备结肠炎模型后,采用皮下注射同型半胱氨酸(0.03μmol·kg-1,每天2次,间隔8 h,连续30 d)造成HHcy模型。实验结束后通过高效液相荧光法(HPLC-FD)检测血浆和结肠粘膜Hcy水平,进行DAI评分并取结肠组织进行HE染色评分,结肠组织匀浆检测MPO活性。结果与正常对照组比较,TNBS模型对照组大鼠DAI和HI评分增高(P<0.01),结肠组织MPO活性增高(P<0.01)。与TNBS模型对照组比较,TNBS模型+Hcy注射组大鼠血浆和结肠粘膜Hcy水平明显增高(P<0.01),DAI和HI评分明显增高(P<0.01),结肠组织MPO活性明显增高(P<0.01)。结论皮下注射同型半胱氨酸可以建立具备高同型半胱氨酸血症的实验性结肠炎模型,可用于探讨Hcy在结肠炎中的作用机制。     

N-乙酰半胱氨酸对葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎的保护作用

目的观察N-乙酰半胱氨酸(NAC)灌肠对小鼠急性溃疡性结肠炎的作用。方法5%葡聚糖硫酸钠(DSS)自由饮用7 d诱导小鼠急性溃疡性结肠炎,同时予以生理盐水(NS)、5-氨基水杨酸(5-ASA)、NAC保留灌肠,观察小鼠体重、粪便性状、隐血便血,计算疾病活动度(DAI)积分,检测结肠长度、结肠过氧化物酶(MPO)活性、血清过氧化物歧化酶(SOD)活力和丙二醛(MDA)含量及肠黏膜病理改变。结果NAC组小鼠隐血、便血、体重下降、DAI积分、病理改变等均好于模型组、NS组(P<0.05),与5-ASA组疗效相似。NAC组SOD活力高于模型组,MPO活性、MDA含量则低于模型组(P<0.05)。结论NAC对DSS诱导的小鼠急性溃疡性结肠炎黏膜损伤有保护作用,其机制可能与抗氧化应激有关。     

IL-21／STAT3通路在溃疡性结肠炎小鼠发病中的表达及其意义

目的探讨IL-21／STAT3通路在溃疡性结肠炎（ulcerativecolitis,UC）小鼠肠道中的表达及其作用。方法20只BALB／C小鼠随机分为正常组和DSS模型组各10只。小鼠UC模型用葡聚糖硫酸钠dextransodiumsulphate,DSS）诱导,观察小鼠疾病活动指数（DAI）评分、病变结肠常规病理切片,采用ELISA法检测血清IL-6、IL-21含量,免疫组化检测结肠STAT3表达。结果模型组小鼠DAI评分、血清IL-6和IL-21含量、结肠病理评分、黏膜STAT3表达水平均较正常组高,差异有显著性（P〈0．05）。结论溃疡性结肠炎小鼠血清炎症介质及肠道STAT3表达水平明显升高,推测IL-21／STAT3通路可能参与UC小鼠肠道炎症损伤,阻断病变肠道IL-21／STAT3通路,减少STAT3蛋白表达或许是治疗UC的作用靶点。     

肠上皮Metrnl基因敲除对肠道菌群以及溃疡性结肠炎的影响

目的 探讨肠上皮Metrnl对葡聚糖硫酸钠盐（DSS）诱导的溃疡性结肠炎小鼠模型的作用及其对肠道菌群调节机制的影响。方法 应用不同浓度的DSS（3%和1%）对C57 小鼠进行造模,确定实验条件。给予肠上皮Metrnl特异性敲除小鼠（Metrnl^(-/-)）及其对照小鼠（Metrnl^(+/+)）3%DSS造模5 d,观察模型小鼠的生存时间、体重、疾病评分（DAI）、结肠长度以及结肠组织切片病理学变化等指标。使用16S核糖体RNA基因测序技术检测肠道菌群组成。结果 与1%DSS相比,3% DSS可显著缩短C57小鼠的生存时间（P<0.05）,降低体重（P<0.05）,增加DAI评分（P<0.05）,缩短结肠长度（P<0.05）,增加病理学评分（P<0.05）。给予3%DSS造模5 d后,与对照组Metrnl^(+/+)小鼠相比,Metrnl^(-/-)小鼠体重下降更多（P<0.05）,DAI评分更高（P<0.05）,结肠长度更短（P<0.05）以及病理学评分更高（P<0.05）。检测16S核糖体RNA结果显示,Metrnl^(-/-)小鼠的肠道菌群多样性下降,拟杆菌（Bacteroidetes）和变形杆菌（Proteobacteria）显著降低,而厚壁菌（Firmicutes）显著升高。结论 Metrnl对3%DSS诱导的溃疡性结肠炎小鼠有保护作用,该作用可能与Metrnl对肠道菌群的调节作用相关。     

A protective effect of melatonin on intestinal permeability is induced by diclofenac via regulation of mitochondrial function in mice

Aim:

This study investigated the effect of intragastrically administered melatonin on intestinal mucosal permeability induced by diclofenac in mice.

Methods:

Intestinal mucosal permeability was induced in mice by intragastric administration of diclofenac (2.5 mg/kg). Melatonin was given intragastrically (10 mg/kg) once per day for 3 d after diclofenac administration. The small intestine was examined macroscopically and microscopically for pathologic injury to the intestinal mucosa. Intestinal mucosal permeability was evaluated by Evans blue and FITC-dextran methods. Mitochondrial functional parameters, including mitochondrial membrane potential, mitochondrial ATPase and succinate dehydrogenase (SDH) activity, were assessed. The malondialdehyde (MDA) and myeloperoxidase (MPO) levels were determined from small intestinal mucosal homogenates.

Results:

As compared with control mice, the permeability, pathologic score, MDA and MPO levels and ulceration of the intestinal mucosa were increased significantly by diclofenac treatment, and a broadened junctional complex and enlarged intercellular space were observed by transmission electron microscopy (TEM). Melatonin treatment significantly reduced the intestinal mucosal permeability, pathologic score, MDA, and MPO levels and ulceration of the intestinal mucosa. By TEM, the small intestine villus morphology and intercellular spaces were nearly normal in melatonin-treated mice. At the level of the mitochondria, melatonin treatment significantly restored the activities of ATPase and SDH.

Conclusion:

The intestinal damage and increased intestinal permeability induced by diclofenac in mice was limited by melatonin; moreover, melatonin preserved several aspects of mitochondrial function.     

Myosin Light Chain Kinase Inhibition: Correction of Increased Intestinal Epithelial Permeability <Emphasis Type="BoldItalic">In Vitro</Emphasis>

Purpose To examine whether myosin light chain kinase (MLCK) inhibitors can reduce intestinal epithelial permeability increases in vitro. Materials and Methods Isolated rat, mouse and human colonic tissue mucosae and Caco-2 monolayers were exposed to cytochalasin D (cD) and sodium caprate (C₁₀), in the absence and presence of the MLCK inhibitors, ML-9 and D PIK. Transepithelial electrical resistance (TEER) and Papp of [¹⁴C]-mannitol or FITC-dextran 4000 (FD-4) were measured. Western blots were used to measure MLC phosphorylation. Results Increases in Papp of [¹⁴C]-mannitol and decreases in TEER were induced by tight junction openers. These changes were attenuated by ML-9. D-PIK offset the FD-4 Papp increase induced by C₁₀ in Caco-2 only, while ML-9 and PIK inhibited MLC directly. cD induced constriction of peri-junctional actin in Caco-2 monolayers, but this was prevented by ML-9. Although mannitol fluxes across colonic mucosae from dextran-sulphate (DSS)-treated mice were higher than control, they were not ameliorated by either ML-9 or PIK in vitro. Conclusions ML-9 inhibits paracellular permeability increases in several intestinal epithelial models. D-PIK reduced stimulated paracellular fluxes in Caco-2 monolayers, but not in tissue. Pre-established increases were not modified by two MLCK inhibitors in a mouse model of IBD.     

The association of minocycline and the probiotic Escherichia coli Nissle 1917 results in an additive beneficial effect in a DSS model of reactivated colitis in mice

Antibiotics have been empirically used for human inflammatory bowel disease, being limited to short periods. Probiotics are able to attenuate intestinal inflammation due to its immunomodulatory properties, being considered as safe when chronically administered. The aim was to test the association of minocycline, a tetracycline with immunomodulatory properties, and the probiotic Escherichia coli Nissle 1917 (EcN) in a mouse model of reactivated colitis. For this purpose, female C57BL/6J mice were assigned to different groups: non-colitic and dextran sodium sulfate (DSS)-control groups (without treatment), minocycline (50 mg/kg/day; p.o.), EcN (5 × 10⁸ CFU/day; p.o.), and minocycline plus EcN treated groups. Colitis was induced by adding DSS in the drinking water (3%) for 5 days; 2 weeks later, colitis was reactivated by subsequent exposure to DSS. The inflammatory status was evaluated daily by a disease activity index (DAI); colonic damage was assessed histologically and biochemically by evaluating mRNA relative expression of different mediators by qPCR. Finally, a microbiological analysis of the colonic contents was performed. Minocycline and EcN exerted intestinal anti-inflammatory effect and attenuated the reactivation of the colitis, as shown by the reduced DAI values, being these effects greater when combining both treatments. This was evidenced histologically and biochemically, by reduced expression of TNFα, IL-1β, IL-2, MIP-2, MCP-1, ICAM-1, iNOS and MMP-9, together with increased MUC-3 and ZO-1 expression. Finally, the altered microbiota composition of colitic mice was partially restored after the different treatments. In conclusion, EcN supplementation to minocycline treatment improves the recovery of the intestinal damage and prevents the reactivation of experimental colitis.     

The association of minocycline and the probiotic Escherichia coli Nissle 1917 results in an additive beneficial effect in a DSS model of reactivated colitis in mice

Antibiotics have been empirically used for human inflammatory bowel disease, being limited to short periods. Probiotics are able to attenuate intestinal inflammation due to its immunomodulatory properties, being considered as safe when chronically administered. The aim was to test the association of minocycline, a tetracycline with immunomodulatory properties, and the probiotic Escherichia coli Nissle 1917 (EcN) in a mouse model of reactivated colitis. For this purpose, female C57BL/6J mice were assigned to different groups: non-colitic and dextran sodium sulfate (DSS)-control groups (without treatment), minocycline (50 mg/kg/day; p.o.), EcN (5 × 10⁸ CFU/day; p.o.), and minocycline plus EcN treated groups. Colitis was induced by adding DSS in the drinking water (3%) for 5 days; 2 weeks later, colitis was reactivated by subsequent exposure to DSS. The inflammatory status was evaluated daily by a disease activity index (DAI); colonic damage was assessed histologically and biochemically by evaluating mRNA relative expression of different mediators by qPCR. Finally, a microbiological analysis of the colonic contents was performed. Minocycline and EcN exerted intestinal anti-inflammatory effect and attenuated the reactivation of the colitis, as shown by the reduced DAI values, being these effects greater when combining both treatments. This was evidenced histologically and biochemically, by reduced expression of TNFα, IL-1β, IL-2, MIP-2, MCP-1, ICAM-1, iNOS and MMP-9, together with increased MUC-3 and ZO-1 expression. Finally, the altered microbiota composition of colitic mice was partially restored after the different treatments. In conclusion, EcN supplementation to minocycline treatment improves the recovery of the intestinal damage and prevents the reactivation of experimental colitis.     

Tacrolimus ameliorates dextran sulfate sodium-induced colitis in mice: implication of interferon-γ and interleukin-1β suppression

We investigated the effect of tacrolimus, a calcineurin inhibitor, on dextran sulfate sodium (DSS)-induced colitis. After inducing colitis in C57BL/6 mice by administering DSS solution as drinking water for 7 d, the animals were treated with tacrolimus. Severity of colonic inflammation was evaluated based on colon weight per unit length. Levels of cytokines (interferon (IFN)-γ, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-12, and tumor necrosis factor (TNF)-α) released from isolated inflamed colons of mice treated with tacrolimus or vehicle were also measured. Treatment with tacrolimus for 14 d reduced the colon weight per unit length and suppressed the release of IFN-γ and IL-1β, but not other cytokines, in inflamed colons of colitic mice compared with vehicle-treated mice. A positive correlation was noted between colon weight per unit length and released level of IFN-γ or IL-1β. The release of IFN-γ and IL-1β was also suppressed after single dosing with tacrolimus to colitic mice. Taken together, these results suggested that tacrolimus ameliorated DSS-induced colitis by suppressing release of IFN-γ and IL-1β from inflamed colon.     

盐酸小檗碱对溃疡性结肠炎小鼠结肠黏膜机械屏障的保护作用

目的 观察盐酸小檗碱（berberine hydrochloride,BBR）对溃疡性结肠炎（ulcerative colitis,UC）小鼠结肠黏膜机械屏障的干预作用,探讨其治疗UC的可能机制。方法 BALB/c小鼠随机分为空白对照组,模型对照组,BBR低、中、高剂量组和柳氮磺吡啶阳性对照组。采用右旋葡聚糖硫酸钠（DSS）法复制UC小鼠模型后,灌胃给药7 d。每天观察小鼠一般情况并评估疾病活动指数（DAI）;末次给药后解剖取结肠组织,切片、HE染色观察组织的病理学变化;Western blot法测定肠道干细胞标志物LGR-5和TERT的含量;免疫组化SP法检测紧密连接蛋白claudin-1、occludin和ZO-1的表达水平。结果 与模型对照组比较,BBR治疗组小鼠结肠炎的临床表现和结肠组织病理表现明显改善,DAI评分下降;经中、高剂量BBR治疗后,UC小鼠结肠组织中LGR-5、TERT和紧密连接蛋白claudin-1、occludin和ZO-1的表达水平均较模型对照组明显升高,差异有统计学意义。结论 BBR能有效减轻UC小鼠的结肠炎症,其机制可能与其抑制肠道干细胞和紧密连接蛋白的破坏、保护肠黏膜机械屏障稳态有关。