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1.
目的:构建2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)以及恶唑酮(oxazolone,OXZ)诱导的小鼠溃疡性结肠炎(ulcerative colitis,UC)模型。方法:120只昆明小鼠()随机分为4组(n=30)。Ⅰ组、Ⅱ组参照Morris及Walter的方法制备小鼠TNBS模型:Ⅰ组(TNBS溶剂对照组),50%乙醇0.1 m L灌肠;Ⅱ组(TNBS模型组),0.6%TNBS溶液0.1 m L灌肠;两组灌肠给药一次后在d 1、d 2、d 3、d 5、d 7每组处死6只。Ⅲ组、Ⅳ组参照Heller方法制备小鼠OXZ模型:Ⅲ组(OXZ溶剂对照组),皮肤涂抹100%乙醇0.1 m L,每天一次,连续2 d,d 7以50%乙醇0.1 m L灌肠;Ⅳ组(OXZ模型组),皮肤涂抹1%OXZ溶液(100%乙醇溶解)0.1 m L每天一次,连续2 d(致敏),d 7以0.5%OXZ(50%乙醇溶解)0.1 m L灌肠;两组灌肠给药一次后在d 1~d 5每天处死6只小鼠。观察Ⅰ~Ⅳ组小鼠疾病活动指数(disease activity index,DAI)、结肠组织大体损伤指数(colon macroscopic damage index,CMDI)和病理组织学评分(histopathological score,HPS),并检测小鼠结肠组织中髓过氧化物酶(myeloperoxidase,MPO)、白细胞介素-4(interleukin-4,IL-4)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平。结果:两种模型组小鼠DAI,CMDI和HPS均较对照组有明显改变;TNBS和OXZ诱导的结肠炎均可导致MPO明显上升,TNBS结肠炎TNF-α明显上升,OXZ结肠炎IL-4明显下降。结论:TNBS及OXZ均能诱导小鼠溃疡性结肠炎模型。两种模型各有特点,其中TNBS诱导的小鼠溃疡性结肠炎以辅助性T1(helper1,Th1)型炎症反应为主,OXZ诱导的小鼠溃疡性结肠炎以辅助性T2(helper2,Th2)型炎症反应为主。  相似文献   

2.
本研究基于脂多糖(LPS)诱导的小鼠脓毒症模型,探讨莲心碱(LIE)对脓毒症中脾损伤的潜在保护作用。脓毒症常伴有炎症、氧化应激和细胞凋亡,并将导致身体器官功能障碍,对脾脏损伤尤甚。将30只小鼠随机分为5组(n=6):对照组、LPS(10 mg/kg)、LIE(10 mg/kg)+LPS、LIE(20 mg/kg)+LPS和LIE(40 mg/kg)+LPS。脾脏损伤通过苏木精–伊红染色(H&E)确定;采用定量聚合酶链反应(q PCR)检测脾脏组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10、IL-1β和一氧化氮合酶(i NOS)的转录水平。同时对包括丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)在内的氧化应激指标进行测定;采用TUNEL法检测细胞凋亡水平。结果显示,LIE可减轻脾脏组织病理学损伤并抑制细胞凋亡,显著降低促炎因子TNF-α、IL-6、IL-1β和i NOS的转录水平,并增加抗炎因子IL-10的表达。此外,LIE预处理后可降低MDA脂质过氧化指标,增强CAT、SOD和GSHPx的抗氧化活性...  相似文献   

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Objective: To investigate the effect of activated protein C (APC) on inflammatory responses in human umbilical vein endothelial cells (HUVEC) stimulated with lipopolysaccharide (LPS). Methods: The second passage of collagenase digested HUVEC was divided into the following groups: serum free medium control group (SFM control), phosphate buffer solution control group (PBS control), LPS group with final concentration of 1 μg/ml (LPS group), APC group with final concentration of 7 μg/ml, Pre-APC group (APC pretreatment for 30 min prior to LPS challenge), and Post-APC group (APC administration 30 min after LPS challenge). Supernatant was harvested at 0, 4, 8, 12 and 24 h after LPS challenge. Interleukin-6 (IL-6) and Interleukin-8 (IL-8) levels were analyzed with ELISA. Cells were harvested at 24 h after LPS challenge, and total RNA was extracted. Messenger RNA levels for IL-6 and IL-8 were semi-quantitatively determined by RT-PCR. Results: Compared with control group, IL-6 and IL-8 levels steadily increased 4 to 24 h after LPS stimulation. APC treatment could increase LPS-induced IL-6 and IL-8 production. The mRNA levels of IL-6 and IL-8 exhibited a similar change. Conclusion: APC can further increase the level of IL-6 and IL-8 induced by LPS. The effect of these elevated cytokines is still under investigation.  相似文献   

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探讨肺内细菌感染患者血清及肺泡灌洗液(BALF)中白介素-8(IL-8)、白介素-6(IL-6)、肿瘤坏死因子α(TNFα)与炎症的相关性及在炎症反应中的作用及意义.血清及BALF中的IL-8及IL6用ELISA法检测、TNFα用RIA法检测,计算BALF中的中性粒细胞(PMN)及肺泡巨噬细胞(AMs)数量.结果:感染组治疗前血清及BALF中IL-8、IL-6、TNFα,BALF中的细胞总数、PMN,AMs比感染组治疗后、哮喘组及对照组明显增高(P〈0.01).革兰阴性菌(G菌)感染者BALF中IL-8、IL-6,TBFα比革兰阳性菌(G+菌)感染者高(P〈0.01).感染组BALF中的IL-8含量与PMN、AMs呈显著正相关(r=0.8671,r=0.6894,P〈0.01).感染组BALF中的IL-6、TNFα含量与AMs呈显著正相关(r=0.8194,r=0.7891,P〈0.01).结论:体内的IL-8、IL—6、TNFα参与肺内细菌感染的炎症反应,检测血清及BALF中的IL-8、IL-6、TNFα对评价肺内细菌感染的病情、病因诊断、估计预后有一定意义.  相似文献   

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应用免疫组化SP法对94例乳腺癌、41例乳腺良性病变和10例正常乳腺组织进行了细胞粘附分子CD_(15)检测。结果发现,CD_(15)。在乳腺癌和乳腺良性病变的阳性率分别为79.8%和58.3%,两者有显著差异性(P<0 01),10例正常乳腺组织仅4例呈CD_(15)弱阳性反应。CD_(15)表达与患者年龄和肿瘤大小无显著关系。CD_(15)表达阳性率在浸润性导管癌中显著高于单纯癌(P<0.05),组织学Ⅱ~Ⅲ级显著高于Ⅰ级者(P<0.05),临床Ⅲ一Ⅳ期显著高于Ⅰ期和Ⅱ期者(P<0.05),淋巴结转移阳性组显著高于阴性组(P<0.01)。在一组原发部位和淋巴结转移性乳腺癌配对标本中,CD_(15)表达无明显差异性。CD_(15)阳性的乳腺癌Cath-D和c-erbB-2的表达阳性率均显著高于CD_(15)阴性者(P<0.001)。结果提示,CD_(15)的表达与乳腺癌的发生发展、浸润转移及预后有密切关系。  相似文献   

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背景:肠道菌群在动物的新陈代谢、生长发育和维持健康过程中发挥了重要作用,调节机体免疫功能,使之处于平衡状态,从而避免或减少免疫相关疾病的发生。一旦肠道菌群紊乱将会导致机体出现内毒素血症,此时可以通过粪菌移植来调节小鼠的肠道菌群紊乱并且抑制由于肠道菌群紊产生的内毒素血症等相关炎症。方法:将30只健康小鼠随机分为3组,每组10只。在相同情况下置于普通动物饲养室饲养一周后,空白组10只小鼠饲喂普通日粮,对照组10只小鼠添加抗生素,实验组10只小鼠在饲喂抗生素后进行粪菌移植;上述小鼠血清采用ELISA法检测TNF-α、IL-1β、IL-6、IL-10以及LPS;对其中部分小鼠肠道进行基于16S rDNA基因的微生物多样性检测。结果:对照组小鼠体内的炎性因子LPS、TNF-α、IL-6较空白组组显著升高(P<0.05),而IL-10、IL-1β的数值显著低于空白组(p<0.05);进行粪菌移植后,实验组小鼠体内的促炎因子浓度明显下降,实验组小鼠血清中LPS、TNF-α、IL-6的水平显著低于治疗前(p<0.05);IL-10、IL-1β的水平高于治疗前(p<0.05);1...  相似文献   

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目的:研究和探讨梓醇对脂多糖(LPS)诱导的牛子宫内膜上皮细胞和小鼠子宫内膜炎的保护机制。创新点:首次证明梓醇对LPS刺激的牛子宫内膜上皮细胞炎症和LPS诱导的小鼠子宫内膜炎具有保护作用,其保护机制与抑制Toll样受体4/核因子κB(TLR4/NF-κB)炎症信号通路有关。方法:通过LPS的诱导,分别建立牛子宫内膜上皮细胞体外炎症模型和小鼠子宫内膜体内炎症模型,设置不同梓醇作用浓度梯度,采用酶联免疫吸附测定法(ELISA)、实时荧光定量聚合酶链式反应(q RT-PCR)、蛋白免疫印迹(western blot)和免疫荧光技术检测TLR4/NF-κB信号通路及其下游炎症因子的表达。结论:梓醇可以显著抑制TLR4和p65 NF-κB信号通路的表达,降低炎性因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)的水平以及趋化因子CXCL8和CXCL5的表达,同时降低子宫组织髓过氧化物酶水平。通过在体内外炎症模型中加入梓醇,可以显著降低牛子宫内膜上皮细胞的炎症反应,有效保护小鼠体内子宫内膜的组织损伤。  相似文献   

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NLRP3炎症小体在炎症反应中发挥重要作用.为了探讨Kv1.3钾离子通道阻断剂对NLRP3炎症小体活化的抑制作用.通过体外培养THP-1细胞,佛波酯诱导其分化为巨噬细胞,LPS联合ATP刺激,试验组提前加入Kv1.3通道阻断剂ADWX-1和不同浓度的PAP-1干预,ELISA检测上清中炎性细胞因子IL-1β分泌,Wes...  相似文献   

9.
目的:以园蓝为研究材料,鉴定园蓝花青素提取物(GBBAEs)中的功能性成分的结构,建立脂多糖(LPS)诱导的体外炎症模型,并评价其抗炎作用和初步机制。创新点:首次探究了蓝莓花青素对建立的LPS诱导体外炎症模型的营养干预作用,并初步探究了发挥抗炎机制的作用通路。方法:将RAW 264.7细胞分为对照组(不作处理)和实验组(1μg/ml LPS刺激建模)。实验组进一步分为3个不同浓度组:400μg/ml GBBAEs组、800μg/ml GBBAEs组和1200μg/ml GBBAEs组。用酶联免疫吸附测定(ELISA)试剂盒检测一氧化氮(NO)、前列腺素E2(PGE2)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、干扰素γ(INF-γ)等炎症因子的释放量;用实时定量聚合酶链反应(RT-PCR)分析IL-1β、IL-6、TNF-α、环氧合酶-2(COX-2)及单核细胞趋化蛋白-1(MCP-1)的炎症相关基因m RNA的表达水平;用蛋白质印迹法(Western blot法)测定相关炎症蛋白COX-2和NF-κBp65表达水平。结论:试验结果表明,通过ELISA法测定GBBAEs可以显著性抑制NO、PGE2、IL-1β、IL-6、INF-γ等炎症因子的释放;RT-PCR分析阐明在LPS诱导的单核-巨噬细胞RAW 264.7中,GBBAEs可以显著性抑制IL-1β、IL-6、TNF-α、COX-2及MCP-1的炎症相关基因m RNA的表达水平。此外,Western blot法进一步显示GBBAEs对相关炎症蛋白COX-2和NF-κBp65表达具有明显抑制作用,进一步证实GBBAEs通过NF-κB机制通路来发挥抗炎作用。  相似文献   

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采用原位合成法和后嫁接法将[SiPMIm] Cl固载在SBA-15材料上,制得SBA-15固载离子液体催化剂SBA-15-[SiPMIm] Cl.用N2吸附-脱附、透射电镜、小角X射线衍射和红外光谱等手段表征催化剂,并考察其在CO2与环氧丙烷(PO)环加成合成碳酸丙烯酯(PC)反应中的催化性能.结果表明:固载等量离子液体后,原位合成法制备的SBA-15固载离子液体催化剂体系对CO2环加成合成PC反应的催化活性优于后嫁接法制备的SBA-15固载离子液体催化剂体系.在最优工艺条件下,PO的转化率高达52.8%,且反应后催化剂经过滤即可分离回收利用,多次使用仍保持较高的反应活性.  相似文献   

11.
Fulminant hepatic failure is a severe clinical condition associated with extremely poor outcomes and high mortality. A number of studies have demonstrated the ability of plasma transfusion to successfully treat fulminant hepatic failure, but the underlying mechanisms are not well understood. The aim of the present study is to define the mechanisms of plasma transfusion treatment in lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced mice. LPS/D-GalN treatment in mice causes significant hepatic failure, including increasing serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, histopathological changes in centrilobular necrosis and inflammatory cells, and the up-regulation of inflammation (tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)). When LPS/D-GaIN-induced mice were treated with plasma, these changes were halted. Results showed that plasma transfusion significantly reduced mortality, and decreased the levels of AST, ALT, and inflammation factors such as TNF-α and IL-6. The expression levels of cleaved Caspase-3, BAX, and p53 were down-regulated and Bcl-2 was up-regulated, suggesting that plasma can reduce LPS/D-GalN-induced apoptosis. The protective mechanism of plasma against LPS/D-GalN-induced fulminant hepatic failure is related to the inhibition of the inflammatory response and the reduction in apoptosis through the down-regulation of the p53-induced apoptotic pathway.  相似文献   

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In vitro study of immunosuppressive effect of apoptotic cells   总被引:3,自引:0,他引:3  
INTRODUCTION Apoptosis plays an important role in differen- tiation, development and pathophysiological proc- esses such as inflammation, neoplasia and autoim- mune diseases. For a long time, apoptotic cells per se and the clearance of apoptotic cells had been viewed as neutral in immune response. Recently many in- vestigations suggested that apoptotic cells actively regulate the immune response (Voll et al., 1997; Fadok et al., 1998; 2000; Byrne and Reen, 2002). Apoptotic cells release …  相似文献   

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Recent studies revealed that apoptotic cells are actively involved in immunosuppression and anti-inflammation. After being phagocytosed by macrophages, apoptotic cells can actively regulate cytokines secretion from lipopolysaccharide (LPS)-stimulated macrophages in which the secretion of immunosuppressive cytokines such as interleukin-10 (IL-10) is increased while the pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNFα), interleukin-1beta (IL-1β) and leukin-8 (IL-8) are suppressed. In this paper, we first present evidence that phagocytosed apoptotic cells regulate cytokine secretion of LPS-stimulated macrophages, but also inhibit the activation of T lymphocytes stimulated by ConA. These data suggest that apoptotic cells can alter the biological behavior of macrophages which gain immunosuppressive property. Project supported by the National Basic Research Program (973) of China (No. 2003CB515500) and the National Natural Science Foundation of China (No. 30371358)  相似文献   

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Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs) and evaluated their capacity and underlying mechanisms in protecting murine RAW 264.7 cells from lipopolysaccharide (LPS)-stimulated inflammation in vitro. Enzyme-linked immunosorbent assay (ELISA) kit results showed that GBBAEs significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), IL-1β, and interferon-γ (INF-γ). Real-time polymerase chain reaction (PCR) analysis indicated that the mRNA expression levels of IL-6, IL-1β, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and cyclooxygenase 2 (COX-2) were suppressed in LPS-stimulated RAW 264.7 cells. Additionally, Western blot analysis was used to evaluate the relative protein expression levels of COX-2 and nuclear factor-κB p65 (NF-κBp65). All these results suggested the potential use of GBBAEs as a functional food for the treatment of inflammatory diseases.  相似文献   

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本文提取兔角膜角蛋白,利用SDS-PAGE以及IEF/SDS-PAGE技术对其组成进行分析研究。结果表明,兔角幕上皮存在四种含量较高的特异性角蛋白,其分子量和等电点分别为:(Mr68kd,pI7.6);(Mr64kd,PI5.4);(Mr58kd,PI7.4);(Mr55kd,PI5.2).  相似文献   

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INTRODUCTIONChronicinfectionwithHBVaffectsmorethan250millionpeopleworldwide.Therearemorethan120millionchronicHBVcarriersinChina;appro-ximately10percentofthemremaininstateofchronichepatitisandhaveahighriskofdevelop-mentofcirrhosisandhepatocellularcarcinoma.ButthereisnoeffectivemethodtocontrolchronicHBVinfectionatpresent.Recentdataindicatedthatim-munotherapeuticstrategiesstimulatingbothcellularandhumoralimmuneresponsestoHBVantigensareessentialforcuringchronicHBVinfection(ChisariandFe…  相似文献   

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