Role of Nuclear Transcription Factor-κB in Endotoxin-induced Shock in Rats

To investigate the role of NF-κB in endotoxic shock in rats. the model of endotoxinshock rats was induced by intravenous infusion of lipopolysaccharidc (LPS). 1 h. 2 h. 4 h and 6 h after LPS injection, the activation of NF-κB in blood mononuclear cells and the content of TNF-α and IL-6 in plasma was detected by enzyme-linked immunoadsordent assay (ELISA). The level of mean arterial pressure (MAP) and the histopathological changes of lung and liver were also observed. The activation of NF-κB in mononuclear cells increased 1 h after LPS injection and reached its peak 2 h after the injection, and its level was higher than that of normal group. The level of TNF-α was increased 1 h after the infusion and peaked 2 h after the injection, and its level was higher than that of normal group after LPS infusion. The content of IL-6 increased gradually with time. the IL-6 level was higher than that of normal group after LPS injection. MAP was decreased gradually with time and its level was lower than that of normal group after LPS injection. Pathological examination showed that endotoxic shock could cause pulmonary alveolar hemorrhage, edema and infiltration of inflammatory cell in lung tissue and congestion, edema, capillary dilation and inflammatory cell infiltration in liver tissue. It is concluded that NF-κB can up-regulate the expression of TNF-α and IL-6 in plasma and play an important role in endotoxin induced shock in rats.     

呼吸机所致肺损伤肿瘤坏死因子-α的表达及核因子-κB活性的实验研究

目的 研究呼吸机所致肺损伤(VILI)时肿瘤坏死因子-α(TNF-α)的表达及核因子-κB(NF-κB)DNA结合活性的变化,探讨VILI炎症反应的分子生物学机制.方法 应用大潮气量(VT)机械通气建立兔VILI模型.40只雄性新西兰兔随机分为对照组、常规VT组、损伤1 h组、2 h组及4 h组.用酶联免疫吸附法(ELISA)检测肺组织匀浆TNF-α含量,反转录多聚酶链反应(RT-PCR)检测mRNA表达.凝胶电泳迁移率分析法(EMSA)测定NF-κB的活性.同时检测动脉血氧分压(PaO₂)和肺湿质量/干质量比值(W/D)及肺组织病理学检查.结果 1) 损伤4 h组PaO₂较常规VT组显著降低(P<0.05),损伤4 h组W/D较对照组和常规VT组显著升高(P均<0.01).2) 损伤2 h组和损伤4 h组肺组织匀浆中TNF-α含量均显著高于对照组和常规VT组(P均<0.01).各损伤组TNF-α mRNA含量均显著高于对照组和常规VT组(P均<0.01),损伤4 h组高于损伤1 h组(P<0.01)和损伤2 h组(P<0.05).3) 各损伤组NF-κB的DNA结合活性均显著高于对照组和常规VT组(P均<0.01),2 h活性达峰值,4 h仍维持在高水平.结论 TNF-α升高参与了VILI的炎症反应过程,其升高可能与其mRNA表达增高有关.NF-κB的DNA结合活性增高可能参与了VILI时TNF-α的基因转录过程.     

积雪草苷对脂多糖诱导的RAW264.7细胞核转录因子κB活化及炎症反应的作用

目的探讨积雪草苷对脂多糖(LPS)诱导的RAW264.7巨噬细胞核转录因子κB(NF-κB)活化及炎症因子表达的影响。方法用LPS刺激RAW264.7巨噬细胞建立炎症模型,四甲基偶氮唑盐(MTT)法检测低、中、高浓度积雪草苷(终浓度分别为10-7、10-6及10-5mol/mL)对RAW264.7细胞增殖的影响,激光共聚焦显微镜观察积雪草苷对细胞NF-κB核转运的作用,ELISA法检测细胞上清中细胞因子TNF-α、IL-1及IL-10的变化。结果低、中、高浓度积雪草苷对RAW264.7细胞增殖均无显著影响。积雪草苷明显抑制RAW264.7细胞NF-κB活化,抑制促炎因子TNF-α和IL-1的表达,同时上调抗炎因子IL-10的表达。空白组、模型组及积雪草苷低、中、高浓度干预组NF-κB转入细胞核百分率分别为(3.5±1.5)%、(75.7±9.1)%、(66.8±7.1)%、(58.9±9.0)%、(40.1±8.8)%,细胞上清TNF-α浓度分别为(171.12±35.42、1775.45±193.97、1284.63±162.13、1035.22±187.97、598.90±107.73)pg/mL,IL-1浓度为(5.66±0.98、26.93±3.48、22.41±2.84、17.05±1.70、10.64±1.29)ng/mL,IL-10为(25.23±2.17、71.75±8.31、82.82±6.00、98.70±8.84、119.97±9.13)pg/mL。结论积雪草苷可能通过抑制NF-κB信号通路,维持促炎系统与抗炎系统的平衡而发挥抗炎作用。     

胎盘早剥患者外周血单个核细胞中NF-κB的表达研究

目的 探讨外周血单个核细胞(peripheral blood mononuclear cells, PBMCs)中NF-κB活化与TNF-α表达在胎盘早剥中的意义。方法以62例胎盘早剥患者为研究对象、正常孕产妇62例为对照组,Western印迹法测定PBMCs中NF-κB蛋白表达水平,RT-PCR检测血清TNF-α mRNA值的变化,并记录新生儿Apgar评分,进行相关性分析。结果与对照组相比,胎盘早剥患者PBMCs NF-κB表达升高(P＜0.05),TNF-α水平升高(P＜0.05),两指标之间呈显著正相关(P＜0.01),与新生儿Apgar评分呈负相关(P＜0.05)。结论NF-κB和TNF-α参与了胎盘早剥病理生理过程,提示胎盘早剥与炎症反应密切相关。     

地塞米松对家兔内毒素休克的防治作用及其与TNF的关系

目的　探讨预防性或治疗性给予地塞米松对家兔内毒素休克的防治作用及其与肿瘤坏死因子 (TNF)的关系。方法　5 3只健康家兔分为对照组 (n =13 ) ,内毒素休克组 (n =16) ,地塞米松预防组 (n =12 )及治疗组 (n =12 )。地塞米松预防组和治疗组动物于脂多糖 (LPS)输注前 3 0min或输注后 2 0min给予地塞米松 5mg/kg体重 ,观察MABP、动物死亡率、循环血中TNF水平及其它指标的变化。结果　与内毒素休克组相比 ,地塞米松预防和治疗组动物平均动脉压 (MABP)有明显上升 (P <0 .0 5 ,P <0 .0 1) ,死亡率下降 (P <0 .0 5 ,P <0 .0 1) ,循环血中TNF水平的升高受到明显的抑制 (P <0 .0 1)。同时 ,地塞米松预防和治疗能使内毒素休克动物血浆胰高血糖素、葡萄糖、乳酸和 β 葡萄糖醛酸酶 (β G)的升高得到明显改善 (P <0 .0 5 ,P <0 .0 1)。体外实验亦表明地塞米松预防能完全抑制肝Kupffer细胞释放TNF。结论　地塞米松对内毒素休克的防治效果与其直接抑制LPS诱导TNF的产生有密切关系。     

Influence of silencing TRAF6 with shRNA on LPS/TLR4 signaling <Emphasis Type="Italic">in vitro</Emphasis>

This study investigated the influence of silencing TRAF6 with shRNA on lipopolysaccharide(LPS)/toll-like receptor(TLR)-4 signaling pathway in vitro.Four plasmids(pGCsi-TRAF6-shRNA1,2,3,4) containing different shRNA sequences were designed and synthesized.The proliferation of RAW264.7 cells after transfected with these plasmids was measured by MTT assay.Inflammatory cellular models were established by LPS stimulation.Levels of TNF-α,IL-1β and TGF-β1 in the supernatants,mRNA expressions of TRAF6,IL-6 and COX-...     

Role of progesterone in TLR4-MyD88-dependent signaling pathway in pre-eclampsia

The role of progesterone in the Toll-like receptor 4 （TLR4）-MyD88-dependent signaling pathway in pre-eclampsia was studied. Peripheral blood mononuclear cells （PBMCs） from pre-eclampsia （PE） patients were subjected to primary culture, and stimulated with different concentra- tions of progesterone （0, 10^-8, 10^-6, and 10^-4 mol/L）. The mRNA expression of TLR4, MyD88 and nu- clear factor-kappaB （NF-κB） was detected by using real-time PCR. The Ikappa-B protein expression was detected by using Western blotting. The expression of tumor necrosis factor-or （TNF-α and inter- leukin-6 （IL-6） in the supernatant was determined by using ELISA. With the concentrations of proges- terone increasing, the mRNA expression levels of TLR4, MyD88 and NF-κB in 2^△△CT value were sig- nificantly decreased, and the IkappaB protein expression levels were significantly increased. The TNF-α and IL-6 expression showed a downward trend when the progesterone concentration increased, and there were significant differences among all of the groups （P〈0.05）. It was suggested that progesterone can inhibit the TLR4-MyD88-dependent signaling pathway in PE significantly and benefit for the preg- nancy.     

Phosphoinositide 3-Kinase/Akt and Nuclear Factor-KB Are Involved in Staphylococcus Aureus-induced Apoptosis in U937 Cells

Objective To explore the mechanisms involved in Staphylococcus aureus （S. aureus） invading human monocytic U937 cells. Methods S. aureus were added to U937 cells at multiplicity of infections （MOI） of 20：1 for 0, 15, 30, 60, and 90 minutes, respectively. Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI） flow cytometry analysis. Akt and nuclear factor-κB （NF-κB） activities were detected by Western blotting. Results Infection of U937 cells with S. aureus induced rapid cell death in a time-dependent manner, and the cells displayed characteristic features of apoptosis. S. aureus-induced apoptosis was associated with a prominent downregulation of activated （phosphorylated） Akt and NF-κB. The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-κB in a dose-dependent manner. Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells. Conclusions S. aureus can stimulate the apoptosis of U937 ceils. S. aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-κB.     

丹参多酚酸B对动脉粥样硬化大鼠炎症细胞因子的影响

目的:研究丹参多酚酸B对动脉粥样硬化大鼠炎症反应相关细胞因子的影响。方法:8周龄雄性SD大鼠60只,随机分成6组(正常组,模型组,丹参多酚酸B低剂量组、中剂量组、高剂量组,辛伐他汀组),并给予相应的处理。电镜下观察心肌超微结构,并取心肌组织用酶联免疫吸附法(ELISA)检测肿瘤坏死因子α(TNF-α)、白细胞介素1α(IL-1α)、白细胞介素6(IL-6)相关炎性因子。结果:各用药组线粒体肿胀以及线粒体嵴排列紊乱等情况较模型组都有明显好转。模型组心肌TNF-α、IL-1α、IL-6浓度均明显高于正常组(P<0.05);丹参多酚酸B不同浓度组以及辛伐他汀组心肌TNF-α浓度显著低于模型组(P<0.05);丹参多酚酸B不同浓度组以及辛伐他汀组心肌IL-1α浓度均有下降,但其差异无统计学意义(P>0.05);丹参多酚酸B中剂量和高剂量组以及辛伐他汀组心肌IL-6浓度显著低于模型组(P<0.05)。结论:丹参多酚酸B具有降低TNF-α、IL-6分泌的作用,从而抑制动脉粥样硬化的炎症反应。     

胃肠癌患者围手术期应用乌司他丁的临床效果评价

目的：探讨胃肠癌患者围手术期应用乌司他丁的临床效果。方法：胃肠癌手术患者30例为实验组,30例为对照组,检测两组术前1d ,术后1、3、5d的血清TNF-α、IL-6、血ALT、血肌酐及血α1-微球蛋白。结果：术后血清TNF-α、IL-6研究组明显低于对照组（P＜0．05）;术后血ALT、血肌酐及血α1-微球蛋白研究组明显低于对照组（P＜0．05）。结论：胃肠癌患者围手术期适当应用乌司他丁可抑制促炎介质释放,减轻重要器官组织损伤,增加围手术期的安全性。     

柴芩承气汤对急性坏死性胰腺炎大鼠胰腺核因子-κB活化的影响

目的：通过观察柴芩承气汤（Chaiqin Chengqi Decoction,CQCQD）对急性坏死性胰腺炎（acute necrotizing pancreatitis,ANP）大鼠的作用,探讨其治疗机制。方法：将30只SD大鼠随机分成3组：假手术（sham-operated,SO）组、ANP组和CQCQD治疗组。大鼠胆总管内逆向泵入3.5%牛胆酸溶液制备ANP模型。6h后分别行腹腔静脉采血送检,取胰腺组织做病理切片;酶标记免疫吸附测定法检测血浆肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）和白细胞介素-6（interleukin-6,IL-6）;HE染色光学显微镜观察胰腺组织病理情况;免疫组织化学染色SP法检测胰腺核因子-κB（nuclear factor-κB,NF-κB）的活化。结果：ANP组白细胞（white blood cell,WBC）计数和血清淀粉酶（amylase,AMY）值较SO组明显升高（P〈0.05,P〈0.01）,CQCQD组血清WBC计数和AMY水平均较ANP组有所下降（P〈0.05）。ANP组的水肿、炎症浸润、出血、坏死和总积分情况较SO组严重（P〈0.01）;CQCQD组的水肿、炎症浸润、出血和总积分水平均较ANP组降低（P〈0.05）。CQCQD组胰腺NF-κB p65阳性细胞的积分光密度（integraloptical density,IOD）值低于ANP组（P〈0.05）。结论：CQCQD可减少血清TNF-α及IL-6含量,降低胰腺NF-κB的活性,减轻胰腺病理损害。     

大鼠失血性休克后单个核细胞释放细胞因子的变化

目的:探讨失血性休克对单个核细胞释放细胞因子的影响.方法:建立大鼠失血性休克模型,采用抗体双夹心ELISA法检测门静脉血、小肠、脾脏单个核细胞分泌的肿瘤坏死因子(TNF-α)与白介素6(IL-6).结果:在内毒素(LPS)刺激下,休克组动物的小肠与门静脉血单个核细胞释放的TNF-α量在复苏后4 h和24 h低于假手术组动物.结论:失血性休克能使小肠单个核细胞在复苏后4 h和24 h对内毒素的刺激呈现低反应.     

Effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with IL-2 or TNF-α

Objective To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2 ) or tumor necrosis factor-α (TNF-α). Methods Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman’s method. Using the isolated cells cultured and treated with IL-2 or TNF-α, we studied the effects of vitamin E on their proliferation and collagen synthesis through an (3)H-thymidine and (3)H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope. Results Adding IL-2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-α, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-α to reproduce or synthesize collagen. Conclusion Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities.     

细胞因子在大鼠溃疡性结肠炎模型中的表达研究

目的：探讨促炎细胞因子IL-6、TNF-α与抑炎细胞因子IL-10在三硝基苯磺酸（TNBS）/乙醇所致的溃疡性结肠炎（ulcerative colitis,UC）大鼠模型结肠组织中的表达,探讨其在该模型中的作用及意义。方法：雌性SD大鼠随机分为正常对照组及模型组,每组7只。正常对照组不造模,模型组用三硝基苯磺酸（TNBS）/乙醇溶液灌肠复制UC模型。观察两组实验大鼠体质量变化、大体及组织病理学改变,采用酶联免疫吸附法检测细胞因子（IL-6、IL-10、TNF-α）的含量。结果：与正常组比较,模型组大鼠结肠大体形态评分、组织学评分明显升高（0.00±0.00与5.43±1.27,1.29±0.49与6.71±0.95,P〈0.01）;与正常组比较,模型组大鼠结肠组织IL-6、TNF-α表达明显升高（102.13±7.12与188.27±11.65,87.39±6.74与121.51±8.56,P〈0.01）;IL-10表达明显下降（202.97±12.26与71.40±8.28,P〈0.01）。结论：促炎细胞因子IL-6、TNF-α与抑炎细胞因子IL-10的失衡在TNBS诱导的大鼠UC发病中起重要作用。     

原花青素对小鼠脑缺血再灌注损伤的神经保护机制

目的：探讨原花青素对小鼠脑缺血再灌注损伤的神经保护机制。方法：120只昆明小鼠随机分成正常对照组、假手术组、14 d 模型组、14 d 治疗组、28 d 模型组、28 d 治疗组;采用小鼠双侧颈总动脉结扎缺血15 min、再灌注15 min,反复3次构建脑缺血再灌注损伤模型;各治疗组在处死前7 d 连续给予松树皮提取物灌胃治疗,其余各组灌注等量生理盐水,小鼠处死后取各组海马组织用 ELISA 法测定肿瘤坏死因子-α（TNF-α）及白介素-6（IL-6）的表达,westen blot 法检测海马内核因子-κB（NF-κB）的表达。结果：各治疗组脑组织 TNF-α、IL-6及 NF-κB 蛋白表达水平与同时点模型组比较均明显降低,差异有统计学意义（P ＜0．05）。结论：原花青素对脑缺血再灌注损伤小鼠神经保护机制可能是通过降低 NF-κB、TNF-α以及 IL-6表达而实现。     

系统性红斑狼疮患者外周血树突状细胞的变化及意义

目的探讨系统性红斑狼疮(SLE)患者外周血浆细胞树突状细胞(PDC)和髓样树突状细胞的(MDC)的比例以及细胞因子干扰素α(IFN-α)、白介素6(IL-6)、肿瘤坏死因子α(TNF-α)的变化及意义。方法选择38例SLE患者(SLE组)、15例健康志愿者(正常对照组)。采用流式细胞术检测PDC、MDC占淋巴细胞的比例,ELISA法检测外周血IFN-α、IL-6、TNF-α水平。结果①SLE组外周血PDC、MDC比例均显著低于正常对照组。②疾病活动期SLE患者外周血PDC比例显著低于稳定期患者;SLE蛋白尿组外周血PDC比例显著低于非蛋白尿组患者;抗ds-DNA阳性组SLE患者外周血PDC比例显著低于抗ds-DNA阴性组。③PDC比例与血沉、SLE疾病活动指数评分呈负相关,与C3呈正相关。④SLE组与正常对照组相比IFN-α、TNF-α、IL-6水平均增高。结论 SLE患者外周血PDC、MDC比例显著降低,且PDC水平与SLE疾病活动及狼疮肾损害有关。     

大鼠原位肝移植后肿瘤坏死因子-α与供肝损伤的关系

目的观察肿瘤坏死因子-α(TNF-α)在肝移植后早期的活性变化及其与供肝损伤的关系。方法建立大鼠肝移植动物模型,观察其生存期外周血ALT变化、大鼠胆汁分泌量及移植肝门静脉血TNF-α活性。结果供肝冷冻保存4h组及对照组移植大鼠存活期超过50d,而冷冻保存8h组大鼠无一长期存活(均<3d);对照组及4h冷冻保存组胆汁分泌量显著多于8h冷冻保存组(P<0.05),8h组ALT水平显著高于对照组及4h组(P<0.05);8h组移植后TNF-α活性显著升高,且术后2h达高峰,而4h组仅在术后2h轻度升高(P<0.05)。结论肝移植早期TNF-α升高程度与供肝损伤程度有关。TNF-α可能源于移植肝的枯否细胞。     

miR-19a对溃疡性结肠炎的作用机制

目的探讨miR-19a对溃疡性结肠炎的作用机制。方法通过生物信息学分析预测miR-19a可能的靶基因,通过免疫组化及Western blotting技术检测靶基因在溃疡性结肠炎小鼠中表达的变化,并进一步通过绿色荧光蛋白报告载体实验对靶基因进行鉴定。结果生物信息学分析预测miR-19a可能的靶基因是TNF-α,免疫组化及Western blotting显示溃疡性结肠炎小鼠肠道组织TNF-α表达增加,miR-19a可抑制TNF-α-3’UTR-WT报告基因活性,而变异型TNF-α-3’UTR-mut报告基因活性不能被抑制。结论 miR-19a的靶基因为TNF-α,其结合位点为TNF-α3’UTR,miR-19a可能在肠道通过直接调控TNF-α而发挥作用。     

慢性间断缺氧对大鼠脑皮层NF-κB、ICAM-1和TNF-α表达的影响

目的 探索慢性间断性缺氧(CIH)对大鼠额顶部皮质-κB(NF-κB)、细胞间黏附分子-1(ICAM-1)和肿瘤坏死因子-α(TNF-α)表达的影响.方法 制作CIH大鼠模型.将30只SD大鼠均分为CIH组(实验组)、空气模拟对照组及空白对照组.采用免疫组织化学方法检测3组大鼠额顶部皮质NF-κB、ICAM-1和TN...