Ginsenoside Rb1 Inhibits Tumor Necrosis Factor-α-Induced Vascular Cell Adhesion Molecule-1 Expression in Human Endothelial Cells

College of Life Sciences, Zhejiang University, Hangzhou, China.
Biological & Pharmaceutical Bulletin (Impact Factor: 1.83). 11/2008; 31(11):2050-6. DOI: 10.1248/bpb.31.2050
Source: PubMed


We investigated whether ginsenoside Rb1 (Rb1) could block tumor necrosis factor-alpha (TNF-alpha)-induced over-expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and human lung microvascular endothelial cells (HMVECs-L). Cells were treated with various concentrations of TNF-alpha with or without Rb1 pre-treatment for 16 h. The mRNA and protein levels of VCAM-1 were determined with real-time polymerase chain reaction (PCR) and flow cytometry, respectively. Human monocytic THP-1 cells labeled with fluorescent dye (Calcein-AM) was used for the adhesion assay on HUVEC monolayers. Dihydroethidium (DHE) was used to demonstrate in situ levels of superoxide production. JC-1 dye was used to measure changes in mitochondrial membrane potential. Activation of mitogen-activated protein kinase (MAPK) and nuclear factor-kappaB (NF-kappaB) was determined by Bio-Plex immunoassay. TNF-alpha treatment significantly increased the mRNA and protein levels of VCAM-1 in HUVECs in a dose dependent manner. Rb1 pre-treatment effectively blocked the TNF-alpha-induced expression of VCAM-1 mRNA or protein by 80% and 43%, respectively (p<0.01). THP-1 adhesion was also blocked. Furthermore, Rb1 reduced the TNF-alpha-induced increase of superoxide anion production by 41% and inhibited the TNF-alpha-induced decrease of mitochondrial membrane potential by 44% in HUVECs. Rb1 also effectively blocked TNF-alpha-induced activation of p38, c-Jun N-terminal protein kinase, extracellular signal-regulated kinase 1/2 and IkappaBalpha. In conclusion, Rb1 effectively blocked the TNF-alpha-induced over-expression of VCAM-1, increased THP-1 adhesion and over-production of superoxide anion. Furthermore, Rb1 inhibited TNF-alpha-induced MAPKs and NF-kappaB activation. These data suggested that Rb1 might have potential therapeutic effects in controlling inflammation in vascular diseases.

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    • "However, the role of monocyte adhesion to the endothelium in the development of atherosclerotic lesions is still not fully understood (Springer, 1994; van Gils et al., 2013). One of these steps is the presentation of different adhesion molecules like vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1) and members of the Selectin family on the cell surface, which is induced by inflammatory stimuli like TNF-(Chai et al., 2008; Chang et al., 2014). It is known that these adhesion receptors interact with cytoskeletal components, which is assumed to be important for firm cell adhesion (Mestas and Ley, 2008; van der Wal et al., 1992). "
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    ABSTRACT: The membrane anchored ligand ephrinB2 belongs to the broad Eph/ephrin system and is able to activate different Eph receptors. The Eph receptors belong to the huge group of receptor-tyrosine kinases. Eph receptors as well as their corresponding ephrin ligands are cell-membrane attached proteins. Therefore, direct cell-cell contact is essentially for interaction. It is known that ephrinB2 plays a pivotal role in developmental and in tumour angiogenesis. Previous studies point to a crucial role of the EphA4-receptor in the process of monocyte adhesion. Since ephrinB2 is known as an interaction partner of EphA4, the aim of the present study was to investigate a possible interplay of EphA4-receptor with ephrinB2 during monocyte adhesion to the endothelium. As verified by bulk adhesion assays and atomic-force microscopy based single-cell force spectroscopy, temporary stimulation of endothelial cells from different sources with the soluble ligand ephrinB2 increased monocyte adhesion to endothelial cells. The proadhesive effect of ephrinB2 was independent of an active transcription, but is mediated via the Rho signaling pathway with subsequent modulation of the actin cytoskeleton. Furthermore, ephrinB2 mediated its impact on monocyte adhesion via the receptor EphA4 as shown by siRNA-mediated silencing. Interestingly, ephrinB2 was induced by TNF-α treatment. Silencing of ephrinB2 led to a lowering of the TNF-α mediated monocyte adhesion to endothelial cells. Furthermore, immunohistochemical staining of human atherosclerotic plaque revealed expression of ephrinB2 in macrophages. The results of the present study point to a crucial role of ephrinB2 induced EphA4 forward signaling in the context of monocyte adhesion to endothelial cells. This transcription-independent effect is mediated by Rho signaling induced actin-filament polymerization.
    No preview · Article · Nov 2015 · Molecular Immunology
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    • "The impact of TNF-α on monocyte adhesion to the endothelium is well known: TNF-α stimulation of endothelial cells leads to an activation of NFκB followed by a transcriptional activation of several adhesion molecules, like VCAM-1, ICAM-1 and E-Selectin [34] [35]. Besides the transcriptional activation of adhesion molecules, TNF-α is able to induce cytoskeletal changes leading to the clustering of adhesion receptors on the surface of endothelial cells [36], which demonstrates the multifunctional influence of TNF-α on monocyte adhesion. "
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    ABSTRACT: The ligand ephrin Al is more often discussed to play a role in the development of the atherosclerotic plaque and in this context especially in the monocyte adhesion to endothelial cells. As tumor necrosis factor-alpha (TNF-alpha) is known to induce monocyte adhesion to endothelium and ephrin A1 expression, the present study focuses on the involvement of ephrin A1 in TNF-alpha-mediated monocyte adhesion. The analysis of different members of the Eph/ephrin system in TNF-alpha-treated human umbilical vein endothelial cells (HUVEC) revealed that especially ephrinAl was found to be highly regulated by TNF-alpha compared to other members of the Eph family. This effect is also present in arterial endothelial cells from the umbilical artery and from the coronary artery. This regulation is dependent on NF kappa B-activation as shown by the expression of a constitutive-active I kappa B-mutant By using siRNA-mediated silencing and adenoviral overexpression of ephrinA1 in HUVEC, the involvement of ephrinA1 in the TNF-alpha triggered monocyte adhesion to endothelial cells could be demonstrated. In addition, these results could be verified by quantitative adhesion measurement using atomic force microscopy-based single-cell force spectroscopy and under flow conditions. Furthermore, this effect is mediated via the EphA4 receptor. EphrinA1 does not influence the mRNA or protein expression of the adhesion receptors VCAM-1 and ICAM-1 in endothelial cells. However, the surface presentation of these adhesion receptors is modulated in an ephrinA1-dependent manner. In conclusion, these data demonstrate that ephrinA1 plays an important role in the TNF-alpha-mediated adhesion of monocytes to endothelial cells, which might be of great importance in the context of atherosclerosis.
    Full-text · Article · Oct 2014 · Journal of Molecular and Cellular Cardiology
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    • "Saponin components of ginsenosides are divided into two different structural classes: the 20(S)-protopanaxadiol (PD) group of ginsenosides including Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Rg3, and Rh2 and 20(S)-protopanaxatriol (PT) group ginsenosides such as Re, Rf, Rg1, Rg2, and Rh1 [23]. PD ginsenosides including Rb1 [24], Rd [25], Rg3 [26], and Rh2 [27] and PT ginsenosides such as Rg1 [28] [29] and Rh1 [30] have been reported to exhibit both in vivo and in vitro antiinflammatory properties in different studies related to inflammation . Individual ginsenoside, however, due to incomplete pharmacokinetic parameters and unknown toxicities, has rarely been reported for the clinical study so far [31]. "
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    ABSTRACT: Despite a multitude of reports on anti-inflammatory properties of ginseng extracts or individual ginsenosides, data on antiarthritic effect of ginseng saponin preparation with mixed ginsenosides is limited. On the other hand, a combined therapy of safe and inexpensive plant-derived natural products such as ginsenosides can be considered as an alternative to treat arthritis. Our previous in vitro data displayed a strong anti-inflammatory action of red ginseng saponin fraction-A (RGSF-A). We, herein, report a marked antiarthritic property of RGSF-A rich in ginsenoside Rb1, Rc, and Rb2. Collagen-induced arthritic (CIA) mice were treated with RGSF-A or methotrexate (MTX) for 5 weeks. Joint pathology, serum antibody production and leukocye activation, cytokine production in the circulation, lymph nodes, and joints were examined. RGSF-A markedly reduced severity of arthritis, cellular infiltration, and cartilage damage. It suppressed CD3(+)/CD69(+), CD4(+)/CD25(+), CD8(+) T-cell, CD19(+), B220/CD23(+) B-cell, MHCII(+)/CD11c(+), and Gr-1(+)/CD11b(+) cell activations. It further suppressed anti-CII- or anti-RF-IgG/IgM, TNF- α , IL-1 β , IL-17, and IL-6 secretions but stimulated IL-10 levels in the serum, joint, or splenocyte. RGSF-A attenuated arthritis severity, modified leukocyte activations, and restored cytokine imbalances, suggesting that it can be considered as an antiarthritic agent with the capacity to ameliorate the immune and inflammatory responses in CIA mice.
    Full-text · Article · Apr 2014 · Mediators of Inflammation
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