Toll样受体4/核因子-кB信号通路在人颅内动脉瘤形成和破裂过程中的激活

<正>颅内动脉瘤病理过程与免疫炎症反应密不可分,但炎症反应的起源或机制仍不太清楚。核转录因子κB(NF-κB)是炎症细胞因子[如白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、IL-6和IL-8]、趋化因子[单核细胞趋化蛋白-1(MCP-1)]、黏附分子[细胞间黏附分子(ICAM)-1和血管细胞黏附分子(VCAM)-1]的主要调节子。研究证实,NF-κB信号转导参与     

番红花酸对人脐静脉内皮细胞VCAM-1表达和单核-内皮细胞黏附的影响

番红花酸为天然类胡萝卜素类化合物,具有抗氧化、抗动脉粥样硬化(atherosclerosis,AS)等多种药理作用,但其抗AS作用机制未明。本研究采用体外培养人脐静脉内皮细胞,观察番红花酸(0.1、1、10μmol·L-1)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ,0.1μmol·L-1)诱导的血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)表达和单核内皮细胞黏附的影响,并观察番红花酸对核转录因子κB(nuclear factor kappa B,NF-κB)活化的影响。结果显示,番红花酸可明显减少AngⅡ诱导的VCAM-1表达及单核-内皮细胞黏附(P<0.05,P<0.01),降低细胞内活性氧(reactive oxygen species,ROS)水平和NF-κB活化水平(P<0.05,P<0.01)。番红花酸还可明显增强内皮细胞总抗氧化能力(P<0.01)和超氧化物歧化酶活性(P<0.05,P<0.01)。这些结果提示,番红花酸可有效抑制VCAM-1表达和单核-内皮细胞黏附,其机制可能与番红花酸增强内皮细胞抗氧化能力、减少细胞内ROS水平,进而抑制NF-κB活化有关。     

蒙花苷对TNF-α诱导的血管内皮细胞炎症损伤及TLR4/IκBα/NF-κB信号通路的影响

目的 以肿瘤坏死因子(TNF-a)刺激人脐静脉内皮细胞(EA.hy926),体外模拟血管内皮细胞炎症损伤模型,探讨蒙花苷(Buddleoside,BUD)对血管内皮细胞的保护作用和机制。方法 MTT法检测TNF-a和蒙花苷对EA.hy926细胞活性的影响;采用TNF-a(50 ng.mL_-1)刺激EA.hy926细胞,加入不同浓度(5、15和25 mmol.L_-1)蒙花苷处理,DAPI染色法测定EA.hy926细胞与单核细胞(THP-1)的黏附能力;Western Blot法检测细胞中Toll样受体(TLR4)、核因子-kB抑制蛋白(IκB-α)、核因子-kB(NF-kB)、细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)等蛋白表达;RT-PCR法检测ICAM-1、VCAM-1等基因mRNA的表达;ELISA法检测细胞上清液中白介素-1(IL-1)、白介素-6(IL-6)和白介素-8(IL-8)的含量。结果 蒙花苷可显著抑制TNF-a刺激EA.hy926细胞与THP-1细胞间黏附作用,降低EA.hy926细胞分泌的炎症因子IL-1、IL-6和IL-8水平。Western blot和RT-PCR结果表明蒙花苷能减少EA.hy926细胞中TLR4、NF-kB、ICAM-1、VCAM-1的蛋白表达量及ICAM-1、VCAM-1 mRNA的表达量,同时能升高细胞中IκB-α的蛋白表达量。结论 蒙花苷能缓解血管内皮细胞炎症损伤,其作用机制可能主要是通过抑制TLR4/IΚBα/NF-κB信号通路发挥效用。     

大米α-球蛋白肽YGEGSSEEG对TNF-α诱导的血管内皮细胞损伤的影响

目的研究从大米α-球蛋白中获取的活性肽发挥抗动脉粥样硬化功效的机制。方法大米α-球蛋白经体外消化产物,采用外翻肠囊法收集可吸收组分,利用凝胶色谱和半制备高效液相色谱分离纯化,液相质谱串联测定氨基酸序列。利用TNF-α诱导人脐静脉内皮细胞损伤模型来评价肽的功效。检测细胞存活率,筛选肽的合适剂量;流式细胞仪检测细胞凋亡率;蛋白质印迹检测Bcl-2、Bax、p-p38、血管细胞黏附分子及NF-κB信号通路等蛋白表达水平;酶联免疫检测谷胱甘肽过氧化物酶等氧化应激因子含量。结果从可吸收组分中获得肽YGEGSSEEG。50 mg·L~(-1)合成肽能够明显抑制TNF-α引起的细胞凋亡,调控Bcl-2/Bax蛋白表达以及阻碍p38蛋白的磷酸化;同时,明显抑制HUVEC产生黏附分子、阻碍NF-κB信号通路的激活;此外,肽对细胞中的氧化应激水平并无明显影响。结论大米α-球蛋白肽YGEGSSEEG可通过抑制细胞凋亡和降低黏附分子表达,发挥血管内皮细胞保护作用。     

高分子量脂联素在EA.hy926血管内皮细胞中对前炎症因子介导的NF-κB信号通路及其下游因子的影响

目的本研究通过促炎因子肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1对血管内皮细胞株EA.hy926的作用,来探讨高分子量脂联素对EA.hy926中NF-κB活性及其下游因子的影响,以阐明高分子量脂联素在血管内皮细胞炎症反应中的作用。方法构建NF-κB荧光素酶报告质粒,并稳定转染血管内皮细胞株EA.hy926。在TNF-α和IL-1作用之前,用不同浓度(0~8μg/ml)高分子量脂联素处理转染后的血管内皮细胞EA.hy926。NF-κB的活性通过荧光酶报告分析法测定。采用酶联免疫吸附试验(ELISA)法测定细胞培养上清液中TNF-α、IL-1及IL-6的浓度。分别采用实时荧光定量聚合酶链反应(real time-PCR)法、蛋白印迹法(Western blot)在m RNA和蛋白水平上检测血管内皮细胞的细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)和单核细胞趋化蛋白-1(MCP-1)的含量。结果研究表明高分子量脂联素抑制促炎因子TNF-α和IL-1介导的NF-κB活性及其下游因子的表达。结论高分子量脂联素通过抑制NF-κB的活性及其下游因子的表达,抑制前炎症因子介导的炎症反应,从而保护血管内皮细胞。     

辛伐他汀对K562细胞NF-κB信号通路的影响

目的 探讨辛伐他汀是否依赖核因子(NF)-κB信号通路分子的变化诱导K562细胞凋亡.方法 体外培养K562细胞,首先用流式细胞术检测辛伐他汀对K562细胞凋亡的影响,RT-PCR分析NF-κB信号通分子转录水平的变化.然后构建裸鼠K562细胞移植瘤模型,TUNEL法检测辛伐他汀对裸鼠移植的K562细胞晚期凋亡的影响,RT-PCR检测裸鼠体内K562细胞NF-κB信号通路分子转录水平的变化.结果 辛伐他汀在体外能明显引起K562细胞发生凋亡,NF-κB通路大多数分子出现差异表达.不同剂量的辛伐他汀能够诱导裸鼠身上移植的K562细胞发生凋亡,并随药物剂量的增加凋亡率逐渐增高(P＜0.01);辛伐他汀能够引起NF-κB信号通分子在转录水平出现明显差异表达(P＜0.01).结论 辛伐他汀可能通过引起NF-κB信号通路分子的变化来诱导K562细胞凋亡的发生.     

血管细胞黏附分子-1与动脉粥样硬化机制的研究现状北大核心CSCD

动脉粥样硬化(AS)是一种慢性的、以脂质发育为特征的炎症过程。血管细胞黏附分子-1(VCAM-1)是一种炎症因子,表达在炎症环境下的内皮细胞膜上,介导白细胞的滚动和黏附。在AS开始初期,免疫细胞和脂质浸润血管壁形成斑块,VCAM-1在这个过程中发挥了重要的作用。本文对目前AS发生、发展过程中VCAM-1高表达的诱因及其病理机制,以及通过减少VCAM-1表达抗AS的药物进行了综述。     

细胞外基质金属蛋白酶诱导因子在动脉粥样硬化核因子 κB信号通路交叉调控中的研究进展

动脉粥样硬化( Atherosclerosis,AS)是心脑血管疾病发生、发展重要的病理基础。细胞外基质金属蛋白酶诱导因子(Extracellular Matrix Metalloproteinase Inducer,EMMPRIN),作为一种广泛分布的、细胞表面糖基化的跨膜蛋白,是 AS重要的促炎蛋白。然而 EMMPRIN在 AS中的细胞信号通路众多且机制繁杂,现着重就 EMMPRIN围绕核因子 κB(Nuclear Factor κB, NF?κB)及其上下游信号通路交叉调控,诱导动脉粥样硬化形成的研究进展作一综述。     

NF-κB——治疗糖尿病微血管并发症的潜在靶点

糖尿病是一种代谢紊乱性疾病,影响着全球大部分人口。核因子-κB(NF-κB)在糖尿病微血管并发症的发病机制中起着关键作用。持续的高血糖可激活NF-κB,从而触发各种细胞因子、趋化因子和细胞黏附分子的表达。NF-κB促使肿瘤坏死因子-α(TNF-α)、白细胞介素、转化生长因子-β(TGF-β)、Bcl2及其他促炎蛋白和促凋亡基因过度表达,是血管功能障碍的主要危险因素。NF-κB过表达也会使内皮细胞钙化,从而导致内皮功能障碍和微血管并发症的发生。抑制NF-κB炎症通路是未来治疗糖尿病微血管并发症的新靶点。在治疗糖尿病并发症方面已经研究了各种天然和合成的NF-κB抑制剂。最近的临床前和临床试验研究证实NF-κB是治疗糖尿病微血管并发症的潜在靶点。     

基于网络药理学和分子对接探讨复方风湿宁片治疗类风湿关节炎的作用机制

目的 采用网络药理学联合分子对接技术探讨复方风湿宁片治疗类风湿性关节炎的作用机制。方法 通过TCMSP、BATMAN-TCM、Swiss TargetPrediction数据库及文献报道收集复方风湿宁片的活性化学成分靶点,并使用GeneCards数据库检索类风湿关节炎的疾病靶点,将两者取交集,并通过绘制“复方风湿宁片–活性成分–靶点–通路–类风湿关节炎”网络图。对复方风湿宁片中的活性成分与类风湿关节炎靶点进行分子对接。结果 通过网络药理学分析得到复方风湿宁片中有34种活性化学成分,对应1059个靶点,与类风湿关节炎靶点交集356个。通过KEGG富集分析发现复方风湿宁片主要可能在肿瘤坏死因子（TNF）信号通路、白细胞介素-17(IL-17)信号通路、Th17细胞分化、T细胞受体信号通路、Toll样受体（TLR）信号通路、核因子κB(NF-κB)信号通路、Th1和Th2细胞分化、Janus激酶（JAK）-信号传导和转录激活蛋白3(STAT3)信号通路、B细胞受体信号通路和类风湿性关节炎等通路上发挥治疗作用。蛋白激酶B(Akt1)、前列腺素内过氧化物酶2(PTGS2)、丝裂原活化蛋白激酶1(...     

Aqueous extract of Salvia miltiorrhoza regulates adhesion molecule expression of tumor necrosis factor alpha-induced endothelial cells by blocking activation of nuclear factor kappaB

Salvia miltiorrhoza Bunge is a traditional herb medicine often used in China for the treatment of cardiovascular disorders, such as atherosclerosis. The purpose of this study was to examine the effect of aqueous extract of Salvia miltiorrhoza Bunge (ESM) on expression of adhesion molecules in tumor necrosis factor (TNF)alpha-induced endothelial cells. When preincubated with ESM (100, 200, 400 microg/mL) for 18 hours, the adhesion of HL-60 cells to TNFalpha-induced endothelial cells was significantly decreased in a concentration-dependent manner, and down-regulation of adhesion molecules, intracellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), was also observed. The inhibitory effect of ESM on TNFalpha-induced VCAM-1 expression was attenuated by inhibition of intracellular glutathione (GSH) synthesis. In addition, ESM also significantly inhibited TNFalpha-induced translocation of nuclear factor kappaB (NF-kappaB) from cytoplasm to nuclei in endothelial cells. These results demonstrated that inhibition of ESM on the expression of adhesion molecules may result from its blocking activation on NF-kappaB. It may imply one of the mechanisms by which ESM exerts its beneficial effect preventing the progress of atherosclerosis.     

Manassantin a and b isolated fromSaururus chinensis inhibit TNF-α-induced Cell adhesion molecule expression of human umbilical vein endothelial cells

Leukocyte adhesion to the vascular endothelium is a critical initiating step in inflammation and atherosclerosis. We have herein studied the effect of manassantin A (1) and B (2), dineolignans, on interaction of THP-1 monocytic cells and human umbilical vein endothelial cells (HUVEC) and expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in HUVEC. When HUVEC were pretreated with 1 and 2 followed by stimulation with TNF-alpha, adhesion of THP-1 cells to HUVEC decreased in dose-dependent manner with IC50 values of 5 ng/mL and 7 ng/mL, respectively, without cytotoxicity. Also, 1 and 2 inhibited TNF-alpha-induced up-regulation of ICAM-1, VCAM-1 and E-selectin. The present findings suggest that 1 and 2 prevent monocyte adhesion to HUVEC through the inhibition of ICAM-1, VCAM-1 and E-selectin expression stimulated by TNF-alpha, and may imply their usefulness for the prevention of atherosclerosis relevant to endothelial activation.     

Section Review Cardiovascular & Renal: Endothelium and atherosclerosis: monocyte accumulation as a target for therapeutic intervention

The endothelium provides a barrier between blood components and vessel wall cells. Changes in endothelial cell functions are pivotal to the pathogenesis of atherosclerosis. Endothelial cell expression of cell-surface vascular cell adhesion molecule-1 (VCAM-1) and vessel wall production of the monocyte chemoattractant protein-1 (MCP-1) coordinate the adhesion and migration of blood-borne monocytes into focal areas of vasculature during initiation and growth of an atherosclerotic lesion. The entry of monocytes into the vessel wall sets into motion a cascade of events that transforms these blood cells into lipid-laden foam cells, major components of an atherosclerotic lesion. The macrophage-foam cells modulate both endothelial and smooth muscle cell functions assuring growth of the lesion. As the lesion matures, the macrophages affect the stability of the plaque predisposing it to rupture. Therefore, accumulation of macrophages lays the groundwork for clinical sequelae such as plaque rapture, thrombosis and, eventually, myocardial infarction. Thus, endothelial cell dysfunction that results in monocyte recruitment profoundly influences atherosclerosis and provides the rationale for therapeutic intervention. This review discusses the cellular and molecular basis for monocyte accumulation in atherosclerosis and potential endothelial cell sites for pharmacological regulation.     

Halofuginone inhibits LPS-induced attachment of monocytes to HUVECs

Atherosclerosis has become a major cause of mortality for several years, however the underlying mechanism of this disorder is still complicated. Endothelial dysfunction is a hallmark in the beginning of atherosclerosis. Lipopolysaccharides (LPS) is an important risk factor contributing to endothelial dysfunction. This study demonstrates that Halofuginone, an anti-malarial drug, possesses protective effects on human umbilical vein endothelial cells (HUVECs) against LPS-induced endothelial dysfunction. Through this study, we demonstrate that Halofuginone ameliorates LPS-induced attachment of THP-1 cells to HUVECs by inhibiting the expressions of adhesion molecules, including vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Halofuginone also suppresses the production of pro-inflammatory cytokines, including tumor necrosis factor α.(TNF-α), interleukin-1β (IL-1β), and interleukin 6 (IL-6). Furthermore, Halofuginone reduces the overproduction of reactive oxygen species (ROS) by regulating the expression of NADPH oxidase 2 (NOX-2). Mechanistically, we find the protective effects of Halofuginone depend on the expression of Kruppel-like factor 2 (KLF2), which is mediated by extracellular regulated protein kinases 5 (ERK5). Totally, our findings demonstrate that Halofuginone possesses a protective function in endothelial cells, indicating a therapeutic potential to modulate endothelial dysfunction in atherosclerosis.     

Cryptotanshinone attenuates in vitro oxLDL-induced pre-lesional atherosclerotic events

Development of early stage atherosclerosis involves the activation of endothelial cells by oxidized low-density lipoprotein (oxLDL) with subsequent increases in endothelial permeability and expression of adhesion molecules favoring the adherence of monocytes to the endothelium. Cryptotanshinone (CTS), a major compound derived from the Chinese herb Salvia miltiorrhiza, is known for its protective effects against cardiovascular diseases. The aim of this study was to determine whether CTS could prevent the oxLDL-induced early atherosclerotic events. OxLDL (100?μg/mL) was used to increase endothelial permeability and induce monocyte-endothelial cell adhesion in human umbilical vein endothelial cells (HUVECs). Endothelial nitric oxide (NO) concentrations, a permeability-regulating molecule, and expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were measured. Results show that a) endothelial hyperpermeability was suppressed by 94?% (p?相似文献   

Inhibition of TNFα-induced adhesion molecule expression by (Z)-(S)-9-octadecenamide, N-(2-hydroxyethyl,1-methyl)

Inflammation is a primary event in atherogenesis. Oleoylethanolamide (OEA), a naturally occurring fatty-acid ethanolamide, lowers lipid levels in liver and blood through activation of the nuclear receptor, peroxisome proliferator-activated receptor-alpha (PPARα). We designed and synthesized (Z)-(S)-9-octadecenamide, N-(2-hydroxyethyl, 1-methyl) (OPA), an OEA analog. The present study investigated the effect of OPA on the expression of adhesion molecules in human umbilical vein endothelial cells (HUVEC). OPA inhibited expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) stimulated by Tumor Necrosis Factor-α (TNF-α) via activation of PPARα. This inhibition of VCAM-1 and ICAM-1 expression decreased adhesion of monocyte-like cells to stimulated endothelial cells. These results demonstrate that OPA may have anti-inflammatory properties. Our results thus provide new insights into possible future therapeutic approaches to the treatment of atherosclerosis.     

Bisacurone inhibits adhesion of inflammatory monocytes or cancer cells to endothelial cells through down-regulation of VCAM-1 expression

Bisacurone, one of the active compounds of the traditionally used indigenous herb Curcuma longa Linne (Zingiberaceae), has anti-oxidant, anti-inflammatory, and anti-metastatic activities. We studied how the level of vascular cell adhesion molecule-1 (VCAM-1), one of the key molecules in the development of atherosclerosis as well as carcinogenesis and metastasis, might be affected by bisacurone in tumor necrosis factor-alpha (TNF-alpha)-activated human umbilical vein endothelial cells (HUVECs). Bisacurone dose-dependently inhibited TNF-alpha-mediated expression of VCAM-1. It showed significant suppressive effect on ROS generation in response to TNF-alpha stimulation and it blocked nuclear factor-kappa B (NF-kappaB) p65 translocation into the nucleus and phosphorylation of inhibitory factor kappaBalpha (IkappaBalpha). It also inhibited phosphorylation of Akt and PKC, which are upstream in the regulation of VCAM-1 by TNF-alpha. Furthermore, bisacurone decreased U937 monocyte and human oral cancer cell (Hep-2, QLL-I, SCC-15) adhesion to HUVECs stimulated by TNF-alpha, suggesting that it may inhibit the binding of these cells by regulating the expression of critical adhesion molecules by TNF-alpha. Thus, bisacurone may be beneficial in the treatment of inflammatory diseases, such as atherosclerosis, where inflammatory monocytes are involved in their pathology, and, moreover, in the development of tumors.     

Sequence recognition of alpha-LFA-1-derived peptides by ICAM-1 cell receptors: inhibitors of T-cell adhesion

Blocking the T-cell adhesion signal from intercellular adhesion molecule-1/leukocyte function-associated antigen-1 interactions (Signal-2) can suppress the progression of autoimmune diseases (i.e. type-1 diabetes, psoriasis) and prevent allograph rejection. In this study, we determined the active region(s) of cLAB.L peptide [cyclo(1,12)Pen-ITDGEATDSGC] by synthesizing and evaluating the biologic activity of hexapeptides in inhibiting T-cell adhesion. A new heterotypic T-cell adhesion assay was also developed to provide a model for the T-cell adhesion process during lung inflammation. Two hexapeptides, ITDGEA and DGEATD, were found to be more active than the other linear hexapeptides. The cyclic derivative of ITDGEA [i.e. cyclo(1,6)ITDGEA] has similar activity than the parent linear peptide and has lower activity than cLAB.L peptide. Computational-binding experiments were carried out to explain the possible mechanism of binding of these peptides to intercellular adhesion molecule-1. Both ITDGEA and DGEATD bind the same site on intercellular adhesion molecule-1 and they interact with the Gln34 and Gln73 residues on D1 of intercellular adhesion molecule-1. In the future, more potent derivatives of cyclo(1,6)ITDGEA will be designed by utilizing structural and binding studies of the peptide to intercellular adhesion molecule-1. The heterotypic T-cell adhesion to Calu-3 will also be used as another assay to evaluate the selectivity of the designed peptides.