Barrier protective effects of rutin in LPS-induced inflammation in vitro and in vivo

Rutin, an active flavonoid compound, is well known to possess potent antiplatelet, antiviral and antihypertensive properties. In this study, we first investigated the possible barrier protective effects of rutin against pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS) and the associated signaling pathways. The barrier protective activities of rutin were determined by measuring permeability, monocytes adhesion and migration, and activation of pro-inflammatory proteins in LPS-activated HUVECs. We found that rutin inhibited LPS-induced barrier disruption, expression of cell adhesion molecules (CAMs) and adhesion/transendothelial migration of monocytes to human endothelial cells. Rutin also suppressed acetic acid induced-hyperpermeability and carboxymethylcellulose-induced leukocytes migration in vivo. Further studies revealed that rutin suppressed the production of tumor necrosis factor-α (TNF-α) and activation of nuclear factor-κB (NF-κB) by LPS. Collectively, these results suggest that rutin protects vascular barrier integrity by inhibiting hyperpermeability, expression of CAMs, adhesion and migration of leukocytes, thereby endorsing its usefulness as a therapy for vascular inflammatory diseases.     

Vascular barrier protective effects of phlorotannins on HMGB1-mediated proinflammatory responses in vitro and in vivo

The phlorotannins (phloroglucinol, eckol, and dieckol) are active compounds found in Eisenia bicyclis, and have been widely investigated for their antioxidant, anti-tumor, and anti-cancer activities. In this study, we investigated the protective effects of these phlorotannins against pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) and in mice treated by high mobility group box 1 protein (HMGB1), and the signaling pathways involved. The protective activities of the phlorotannins were determined by measuring permeability, leukocyte adhesion and migration, and the activations of pro-inflammatory proteins in HMGB1-activated HUVECs. We found that the phlorotannins inhibited; lipopolysaccharide (LPS)-induced HMGB1 release, HMGB1-mediated barrier disruption, the expressions of cell adhesion molecules (CAMs), and the adhesion/transendothelial migration of leukocytes to human endothelial cells. The phlorotannins also suppressed acetic acid induced-hyperpermeability and carboxymethylcellulose-induced leukocytes migration in vivo. Further studies revealed that the hydroxyl groups on dieckol positively regulated these vascular barrier protective effects. Collectively, these results suggest that phloroglucinol, eckol, and dieckol protect vascular barrier integrity by inhibiting hyperpermeability, the expressions of CAMs, and the adhesion and migration of leukocytes, which confirms their potential usefulnesses for the treatment of vascular inflammatory diseases.     

Anti-septic effects of glyceollins in HMGB1-induced inflammatory responses in vitro and in vivo

The ubiquitous nuclear protein High mobility group box 1 (HMGB1) is released by activated macrophages and human umbilical vein endothelial cells (HUVECs), and functions as a late mediator of experimental sepsis. Glyceollins (GCLs) are active compounds from Aspergillus sojae which have been reported for anti-cancer, anti-diabetes, and anti-inflammatory activities. We investigated here, the antiseptic effects and underlying mechanisms of GCLs against HMGB1-mediated septic responses in HUVECs and mice. According to the results, GCLs effectively inhibited lipopolysaccharide-induced release of HMGB1, and suppressed HMGB1-mediated septic responses, such as hyperpermeability, adhesion and migration of leukocytes, and expression of cell adhesion molecules. In addition, GCLs suppressed the production of tumor necrosis factor-α and interleukin 6 and activation of nuclear factor-κB and extracellular regulated kinases 1/2 by HMGB1. Collectively, these results indicate that GCLs could be a potential therapeutic agent for treatment of various severe vascular inflammatory diseases via inhibition of the HMGB1 signaling pathway.     

Anti-septic effects of pelargonidin on HMGB1-induced responses in vitro and in vivo

A certain nucleosomal protein—high mobility group box-1 (HMGB1)—has recently been established as a late mediator of sepsis, with a relatively wide therapeutic window for pharmacological intervention. Pelargonidin (PEL) is a well-known red pigment found in plants; it has important biological activities that are potentially beneficial for human health. In the present study, we investigated whether PEL can modulate HMGB1-mediated inflammatory responses in human umbilical vein endothelial cells (HUVECs) and in mice. The anti-inflammatory activities of PEL were determined by measuring permeability, leukocyte adhesion and migration, and activation of pro-inflammatory proteins in HMGB1-activated HUVECs and mice, as well as the beneficial effects of PEL on survival rate in the mouse sepsis model. The data showed that PEL had effectively inhibited lipopolysaccharide (LPS)-induced release of HMGB1 and suppressed HMGB1-mediated septic responses, such as hyperpermeability, adhesion and migration of leukocytes, and expression of cell adhesion molecules. Furthermore, PEL inhibited the HMGB1-mediated production of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), as well as the activation of nuclear factor-κB (NF-κB) and extracellular signal-regulated kinases 1 and 2 (ERK1/2). Collectively, these results indicate that PEL could be used to treat various severe vascular inflammatory diseases via the inhibition of the HMGB1 signaling pathway.     

Inhibitory effects of lycopene on HMGB1-mediated pro-inflammatory responses in both cellular and animal models

High mobility group box 1 (HMGB1) mediates proinflammatory responses in inflammatory diseases. Lycopene found in tomatoes and tomato products has anti-oxidant, anti-cancer and antiinflammatory effects. The potential anti-inflammatory roles of lycopene in HMGB1-mediated proinflammatory responses in both primary human umbilical vein endothelial cells (HUVECs) and animal were investigated. The anti-inflammatory effects of lycopene were determined including permeability, monocyte adhesion and migration, and activation of proinflammatory proteins and HMGB1 receptors on HMGB1 activated HUVECs. In the in vivo model, the anti-inflammatory effect of lycopene was assessed by monitoring vascular permeability and migration of leukocytes to the peritoneal cavity of mice injected with lycopene. Lycopene inhibited lipopolysaccharide (LPS)-mediated release of HMGB1, expression of HMGB1-mediated tumor necrosis factor (TNF)-secretory phospholipase A2 (sPLA2)-IIA, and HMGB1-mediated pro-inflammatory signaling responses in endothelial cells. It did this through down-regulation of cell surface expression of cell adhesion molecules (CAMs), HMGB1 receptors, toll-like receptor (TLR)-2, and -4, and receptors for advanced glycation end products (RAGE). These findings suggest that lycopene promotes barrier integrity, inhibits monocyte adhesion and migration to HMGB1 activating HUVECs by blocking activation of proinflammatory cytokines and expression of CAMs and HMGB1 receptors, thereby showing its usefulness as a therapy for vascular inflammatory diseases.     

Ecklonia cava extracts inhibit lipopolysaccharide induced inflammatory responses in human endothelial cells

Ecklonia cava (EC) is a brown alga that evidences radical scavenging, bactericidal, tyrosinase inhibitory and protease inhibitory activities. However, the antiinflammatory effects in human endothelial cells and its molecular mechanism remain poorly understood. In this study, we attempted to determine whether pretreatment with EC extracts induce a significant inhibition of antiinflammatory activities in lipopolysaccharide (LPS) induced human endothelial cells. We found that each EC extract inhibits LPS induced barrier permeability, expression of cell adhesion molecules, monocytes adhesion, and transendothelial migration to human endothelial cells. Further studies revealed that EC extracts suppress the production of tumor necrosis factor-α (TNF-α) and activation of nuclear factor-kappa B (NF-κB). Particularly, the antiinflammatory effects of ethyl acetate (EtOAc) and butanol (n-BuOH) extracts were better than those of other extracts. Collectively, these results suggest that EC extracts possess barrier integrity activity, inhibitory activity on cell adhesion and migration to endothelial cells by blocking the activation of NF-κB expression and production of TNF-α, thereby endorsing its usefulness as therapy for vascular inflammatory diseases.     

Geniposide inhibits high glucose-induced cell adhesion through the NF-KB signaling pathway in human umbilical vein endothelial cells

Aim:

To investigate whether geniposide, an iridoid glucoside extracted from gardenia jasminoides ellis fruits, inhibits cell adhesion to human umbilical vein endothelial cells (HUVECs) induced by high glucose and its underlying mechanisms.

Methods:

HUVECs were isolated from human umbilical cords and cultured. The adhesion of monocytes to HUVECs was determined using fluorescence-labeled monocytes. The mRNA and protein levels of vascular cell adhesion molecule-1 (VCAM-1) and endothelial selectin (E-selectin) were measured using real-time RT-PCR and ELISA. Reactive oxygen species (ROS) production was measured using a fluorescent probe. The amounts of nuclear factor-kappa B (NF-κB) and inhibitory factor of NF-κB (IκB) were determined using Western blot analysis. The translocation of NF-κB from the cytoplasm to the nucleus was determined using immunofluorescence.

Results:

Geniposide (10–20 μmol/L) inhibited high glucose (33 mmol/L)-induced adhesion of monocytes to HUVECs in a dose-dependent manner. This compound (5–40 μmol/L) also inhibited high glucose-induced expression of VCAM-1 and E-selectin at the gene and protein levels. Furthermore, geniposide (5–20 μmol/L) decreased ROS production and prevented IκB degradation in the cytoplasm and NF-κB translocation from the cytoplasm to the nucleus in HUVECs.

Conclusion:

Geniposide inhibits the adhesion of monocytes to HUVECs and the expression of CAMs induced by high glucose, suggesting that the compound may represent a new treatment for diabetic vascular injury. The mechanism underlying this inhibitory effect may be related to the inhibition of ROS overproduction and NF-κB signaling pathway activation by geniposide.     

阿托伐他汀对内皮细胞微粒诱导人脐静脉内皮细胞VCAM-1和ICAM-1表达的影响

目的:探讨阿托伐他汀对内皮细胞微粒(EMPs)诱导的人脐静脉内皮细胞(HUVECs)表达血管细胞粘附分子(VCAM)-1和细胞间粘附分子(ICAM)-1的影响。方法:取生长良好的第4,5代人脐静脉内皮细胞,将细胞分为3大组:对照组、EMPs组、EMPs+阿托伐他汀组。对照组加入培养基,EMPs组以不同浓度的EMPs(0/mL,1×102/mL,1×103/mL,1×104/mL,1×105/mL)与HUVECs共同孵育24 h,EMPs+阿托伐他汀组以不同浓度的阿托伐他汀(0.05,0.1,1.0,10μmol.L-1)与HUVECs作用1 h后,加入105/mL EMPs共同孵育24 h。分别采用实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测VCAM-1和ICAM-1 mRNA和蛋白的表达。结果:HUVECs受EMPs刺激后,VCAM-1和ICAM-1 mRNA及蛋白表达呈浓度依赖性增加,阿托伐他汀可不同程度上抑制EMPs的作用。结论:阿托伐他汀抗动脉粥样硬化作用可能部分与抑制EMPs诱导的内皮细胞VCAM-1和ICAM-1的表达有关。     

姜黄素通过阻断NF-κB减少IL-1β诱导的内皮脂酶表达

目的观察姜黄素对培养的人血管内皮细胞内皮脂酶表达的影响,并探讨其可能的作用机制。方法不同浓度的姜黄素处理HUVEC-12细胞。RT-PCR检测内皮酯酶(endo-thelial lipase,EL)mRNA的表达;Western blot检测核因子-κB抑制因子-α(inhibitor of nuclear factor-κB-α,IκB-α)蛋白的表达;间接免疫荧光检测核因子-κB(nuclear factor-κB,NF-κB)蛋白的活化。结果IL-1β处理HUVEC-12细胞可以明显上调EL mRNA的表达,同时降低胞质蛋白IκB-α的表达水平,激活核转录因子NF-κB,增加胞核蛋白NF-κB的水平。姜黄素预处理HUVEC-12细胞可以抑制IL-1β对EL的上调作用,同时逆转IL-1β诱导的IκB-α蛋白降解和NF-κB活化。结论姜黄素可以通过阻断NF-κB活化减少IL-1β诱导的人血管内皮细胞EL的表达。     

西洛他唑对糖尿病大鼠主动脉VCAM-1、NF-κB及PPARs的影响

目的观察西洛他唑对糖尿病大鼠主动脉血管细胞粘附分子-1(VCAM-1)、核因子(NF)-κB及过氧化物酶增殖体激活受体(PPAR s)的影响,探讨西洛他唑影响VCAM-1表达的上游信号通路。方法腹腔注射链脲佐菌素(65 mg.kg-1)制备糖尿病模型,正常对照组大鼠腹腔注射柠檬酸钠缓冲液(NC,n=8)。随机将成模的32只糖尿病大鼠分为糖尿病模型组(DM,n=12)、高剂量西洛他唑组(GX,27mg.kg-1.d-1,n=10)与低剂量西洛他唑组(DX,9 mg.kg-1.d-1,n=10)。治疗8 wk,采用免疫组织化学法检测各组主动脉组织VCAM-1表达及P65亚基核转位情况;原位杂交和凝胶电泳迁移率改变分析法(EMSA)检测主动脉VCAM-1 mRNA表达及NF-κB-DNA结合活性;RT-PCR或Real-Time PCR检测PPARα、PPARγmRNA表达。结果①与正常组相比,糖尿病大鼠主动脉内皮VCAM-1及其基因表达升高(P<0.01),NF-κB P65核转位及活化增强(P<0.01),PPARγmRNA表达为正常大鼠的1.7倍,而PPARαmRNA表达下降(P<0.01)。②与糖尿病组相比,高剂量西洛他唑治疗组主动脉内皮VCAM-1及其基因表达降低(P<0.01),NF-κB P65核转位及活化被抑制(P<0.01),PPARγmRNA表达比糖尿病大鼠下降73%,而PPARαmRNA则上升(P<0.05)。结论高剂量西洛他唑抑制糖尿病大鼠主动脉内皮表达VCAM-1,这可能与其对PPARs表达、NF-κB核转位及其DNA结合活性的影响有关。     

Anti-inflammatory activities of oleanolic acid on HMGB1 activated HUVECs

As a late mediator of inflammation, high mobility group box 1 (HMGB1) protein up-regulates pro-inflammatory cytokines in several inflammatory diseases. Further, high plasma levels of HMGB1 correlate with poor prognosis and increased mortality in patients with severe inflammation. Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but the effects of OA on HMGB1-mediated pro-inflammatory responses of human endothelial cells is not well-studied. In this study, we investigated this question by monitoring the effect of OA on lipopolysaccharide (LPS)-mediated release of HMGB1 and the HMGB1-mediated modulation of inflammatory responses in human umbilical vein endothelial cells (HUVECs). OA potently inhibited the release of HMGB1 by HUVECs as well as down-regulated HMGB1-dependent adhesion and migration of the monocytic cell line THP-1 to activated HUVECs. OA also down-regulated the cell surface expression of the receptor of HMGB1, thereby inhibiting HMGB1-dependent pro-inflammatory responses by inhibiting activation of nuclear factor-κB (NF-κB) and production of tumor necrosis factor-α (TNF-α) by HMGB1. Given these results, OA showed anti-inflammatory activities and could be a candidate as a therapeutic agent for various inflammatory diseases through the inhibition of the HMGB1 signaling pathway.     

NADPH oxidase and lipid raft-associated redox signaling are required for PCB153-induced upregulation of cell adhesion molecules in human brain endothelial cells

Exposure to persistent organic pollutants, such as polychlorinated biphenyls (PCBs), can lead to chronic inflammation and the development of vascular diseases. Because cell adhesion molecules (CAMs) of the cerebrovascular endothelium regulate infiltration of inflammatory cells into the brain, we have explored the molecular mechanisms by which ortho-substituted polychlorinated biphenyls (PCBs), such as PCB153, can upregulate CAMs in brain endothelial cells. Exposure to PCB153 increased expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), as well as elevated adhesion of leukocytes to brain endothelial cells. These effects were impeded by inhibitors of EGFR, JAKs, or Src activity. In addition, pharmacological inhibition of NADPH oxidase or disruption of lipid rafts by cholesterol depleting agents blocked PCB153-induced phosphorylation of JAK and Src kinases and upregulation of CAMs. In contrast, silencing of caveolin-1 by siRNA interference did not affect upregulation of ICAM-1 and VCAM-1 in brain endothelial cells stimulated by PCB153. Results of the present study indicate that lipid raft-dependent NADPH oxidase/JAK/EGFR signaling mechanisms regulate the expression of CAMs in brain endothelial cells and adhesion of leukocytes to endothelial monolayers. Due to its role in leukocyte infiltration, induction of CAMs may contribute to PCB-induced cerebrovascular disorders and neurotoxic effects in the CNS.     

活化蛋白C通过NF-κB抑制TNF-α介导的炎症反应

目的观察活化蛋白C(APC)是否能够通过核因子-κB(NF-κB)途径抑制TNF-α介导的炎症反应。方法分离培养正常组、TNF-ɑ组、TNF-ɑ+rhAPC组的人脐静脉内皮细胞细胞(HUVECs)。用ELISA法,检测细胞上清液中的细胞内粘附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)及选择素浓度。用逆转录聚合酶链反应、Western bloting技术检测HUVECs NF-κBmRNA、NF-κB蛋白的表达。结果细胞上清液中ICAM-1、VCAM-1及E选择素浓度,TNF-ɑ组较正常组显著升高(均P<0.05);而TNF-ɑ+rhAPC组较TNF-ɑ组显著降低(均P<0.05)。HUVECs NF-κB mRNA、蛋白表达水平,TNF-ɑ组较正常组均显著升高(均P<0.05);而TNF-ɑ+rhAPC组较TNF-ɑ组均显著降低(均P<0.05)。结论 APC通过抑制黏附分子的产生来调节TNF-α介导的炎症反应,其可能是通过NF-κB途径来实现。     

Amelioration of compound 4,4′-diphenylmethane-bis(methyl)carbamate on high mobility group box1-mediated inflammation and oxidant stress responses in human umbilical vein endothelial cells via RAGE/ERK1/2/NF-κB pathway

High mobility group box-1 (HMGB1), a secreted nuclear protein, acts as an inflammatory mediator and has been implicated in pathophysiological damage of diabetic vascular complications. A compound 4,4′-diphenylmethane-bis(methyl) carbamate (CM1) has a protective activity on advanced glycation end products (AGEs)-induced endothelial dysfunction in our previous study. The aim of this study was to investigate whether CM1 could attenuate HMGB1-induced endothelial dysfunction in human umbilical vein endothelial cells (HUVECs), and also elucidate the possible underlying mechanism. The pre-treatment of CM1 (10^? 9 M) could inhibit significantly the migration of macrophages in co-incubation with HUVECs system. HMGB1 stimulated intercellular adhesion molecule-1 (ICAM-1), transforming growth factor-beta1 (TGF-β1) and receptor for advanced glycation end products (RAGE) protein expression in HUVECs, which were inhibited by pretreatment with CM1. Furthermore, it also reduced significantly reactive oxygen species (ROS) generation and inflammatory cytokine interleukin-6 (IL-6) level in co-incubation system. Immunofluorescence and Western blotting assays showed that CM1 could attenuate HMGB1-induced intracellular ERK1/2 and NF-kB activation in HUVECs. Our findings indicated that CM1 attenuated HMGB1-mediated endothelial activation by ameliorating inflammation and oxidant stress responses via RAGE/ERK1/2/NF-κB pathway.     

Peoniflorin prevents the adhesion between inflammatory endothelial cells and leukocytes through inhibiting the activation of MAPKs and NF‐κB

The vascular endothelium can be activated by multiple factors, including lipopolysaccharide (LPS) functioning as a key component of the inflammatory response. Activated endothelium promotes the recruitment of leukocytes mainly by releasing various adhesion molecules and amplifies inflammation via a feedback loop. Peoniflorin, the main active constituent of the roots of Paeonia lactiora Pall., possesses anti‐inammatory, anti‐infective, and anti‐platelet aggregative properties. To elucidate the anti‐inammatory mechanism of peoniflorin, the present study was conducted to address its effects on the adhesion of inflammatory endothelial cells to leukocytes. Peoniflorin substantially reduced adhesion of either human acute monocytic leukemia cells (THP‐1) or human acute promyelocytic leukemia cells (HL‐60) to LPS‐stimulated human umbilical vein endothelial cells (HUVECs). It also markedly down‐regulated the expression of E‐selectin at both the gene and protein levels. However, peoniflorin only slightly reduced expression of intercellular adhesion molecule‐1 (ICAM‐1). Signal pathway analysis indicated that peoniflorin reduced phosphorylation of c‐Jun NH₂‐terminal kinase (JNK) and p38 kinase, as well as the phosphorylation and degradation of IκB‐α in HUVECs. These findings suggest that prevention of the adhesion between inflammatory endothelial cells and leukocytes contributes, at least partially, to the anti‐inflammation action of peoniflorin. The anti‐adhesion mechanisms of peoniflorin were involved in the down‐regulation of the activation of mitogen‐activated protein kinases (MAPKs) and nuclear factor‐kappa B (NF‐κB), and a reduction of adhesion molecule expressions in endothelial cells. Drug Dev Res 2010.© 2010 Wiley‐Liss, Inc.     

Gossypol decreases tumor necrosis factor-α-induced intercellular adhesion molecule-1 expression via suppression of NF-κB activity

Gossypol is a yellowish polyphenolic compound originally from cotton plant, which has been shown to exert a potential for anti-cancer and anti-inflammatory effects. However, its molecular mechanism is not thoroughly understood on breast cancer cells known to highly express intercellular adhesion molecule-1 (ICAM-1) for their adhesion and metastasis. This study aims to investigate the effect of gossypol on tumor necrosis factor (TNF)-α-stimulated ICAM-1 via nuclear factor-kappa B (NF-κB) activity. Gossypol was shown to inhibit TNF-α-induced ICAM-1 expression and U937 cell adhesion to MDA-MB-231 and MCF-7 cells. Additionally, TNF-α-induced MDA-MB-231 cell invasion was blocked in the presence of gossypol. Chromatin immunoprecipitation analysis demonstrated that gossypol blocks NF-κB binding on the ICAM-1 promoter regions. Additionally, TNF-α-induced NF-κB activation was completely suppressed in the presence of gossypol. Gossypol did not directly suppress the binding of NF-κB to the DNA but rather inhibited the nuclear translocation of p65 and p50 via phosphorylation and degradation of IκB. We also found that gossypol suppresses NF-κB activation induced by a wide variety of agents, including taxol, okadaic acid, and phorbol myristate acetate. Taken together, gossypol effectively inhibited TNF-α-induced ICAM-1 expression via the suppression of NF-κB activation and in vitro adhesion and invasion in human breast cancer cells.     

The dipeptidyl peptidase (DPP)-4 inhibitor trelagliptin inhibits IL-1β-induced endothelial inflammation and monocytes attachment

Cardiovascular diseases remain the major cause of death worldwide. Atherosclerosis is recognized as the common ground of cardiovascular diseases. Inflammatory cytokines-induced attachment of monocytes to endothelial cells is a significant event in the progression of atherosclerosis. As a highly selective dipeptidyl peptidase (DPP)-4 inhibitor, trelagliptin is used for the treatment of type 2 diabetes mellitus (T2DM). However, whether trelagliptin possesses an inhibitory effect on endothelial dysfunction and monocyte adhesion is unknown. In the current study, we tested the effect of trelagliptin in endothelial cells. We used human aortic endothelial cells (HAECs) exposed to interleukin (IL)-1β to mimic the microenvironment of atherosclerosis. Our results showed that trelagliptin inhibited the expression of pro-inflammatory chemokines including monocyte chemoattractant protein 1 (MCP-1), CXCL-1, and IL-6. Furthermore, trelagliptin suppressed the expression of adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). Mechanistically, trelagliptin suppressed the activation of activator protein-1 (AP-1) and NF-κB signaling pathways, which modulate the inflammatory process and monocyte adhesion. Collectively, our results showed that trelagliptin had a powerful inhibitory effect on the attachment of monocytes to endothelial cells, indicating that trelagliptin might have a protective effect on cardiovascular diseases such as atherosclerosis.     

参附对脐静脉内皮细胞Toll样受体4/NF-κB途径的影响

目的观察参附注射液对内毒素介导的人脐静脉细胞Toll样受体4/NF-κB途径及细胞间黏附分子、E-选择素表达的影响。方法健康产妇分娩后的新鲜婴儿脐带,应用胶原酶消化,收集内皮细胞进行培养。实验分为对照组、内毒素组、内毒素+参附组。对照组:M199培养基中不加任何物质。内毒素组:M199培养基中加入脂多糖1 mg/L。内毒素+参附组:预先加入参附注射液10 mL/L,30 min后加入脂多糖1 mg/L。干预1h收集细胞,提取核蛋白测定NF-κB活性,干预12 h后收集细胞检测Toll样受体4、细胞间黏附分子1、E-选择素表达水平。结果内毒素组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均显著高于对照组,内毒素+参附组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均明显低于内毒素组。结论参附注射液可以通过抑制NF-κB活化,减弱Toll样受体4/NF-κB途径,抑制了脂多糖介导的黏附分子表达,从而减轻机体的炎症反应。     

The coffee diterpene kahweol inhibits tumor necrosis factor-alpha-induced expression of cell adhesion molecules in human endothelial cells

Endothelial cells produce adhesion molecules after being stimulated with various inflammatory cytokines. These adhesion molecules play an important role in the development of atherogenesis. Recent studies have highlighted the chemoprotective and anti-inflammatory effects of kahweol, a coffee-specific diterpene. This study examined the effects of kahweol on the cytokine-induced monocyte/human endothelial cell interaction, which is a crucial early event in atherogenesis. Kahweol inhibited the adhesion of TNFalpha-induced monocytes to endothelial cells and suppressed the TNFalpha-induced protein and mRNA expression of the cell adhesion molecules, VCAM-1 and ICAM-1. Furthermore, kahweol inhibited the TNFalpha-induced JAK2-PI3K/Akt-NF-kappaB activation pathway in these cells. Overall, kahweol has anti-inflammatory and anti-atherosclerotic activities, which occurs partly by down-regulating the pathway that affects the expression and interaction of the cell adhesion molecules on endothelial cells.