TRIM14 promotes endothelial activation via activating NF-κB signaling pathway

Endothelial activation by proinflammatory cytokines is closely associated to the pathogenesis of atherosclerosis and other vascular diseases;however, the molecular mechanisms controlling endothelial activation are not fully understood. Here we identify TRIM14 as a new positive regulator of endothelial activation via activating NF-κB signal pathway. TRIM14 is highly expressed in human vascular endothelial cells (ECs) and markedly induced by inflammatory stimuli such as TNF-α, IL-1β, and LPS. Overexpression of TRIM14 significantly increased the expression of adhesion molecules such as VCAM-1, ICAM-1, E-selectin, and cytokines such as CCL2, IL-8, CXCL-1, and TNF-α in activated ECs and by which it facilitated monocyte adhesion to ECs. Conversely, knockdown of TRIM14 has opposite effect on endothelial activation. Upon TNF-α stimulation, TRIM14 is recruited to IKK complex via directly binding to NEMO and promotes the phosphorylation of IκBα and p65, which is dependent on its K63-linked ubiquitination. Meanwhile, p65 can directly bind to the promoter regions of human TRIM14 gene and control its mRNA transcription. Finally, TRIM14 protein level is significantly upregulated in mouse and human atheroma compared to normal arteries. Taken together, these results indicate that TRIM14-NF-κB forms a positive feedback loop to enhance EC activation and TRIM14 may be a potential therapeutic target for vascular inflammatory diseases such as atherosclerosis.

The law of anti-VCAM-1 targeted microbubbles adhesion to activated endothelial cells under controlled shear flow

Microbubbles can enhance the detection in noninvasive ultrasound imaging.Recently,targeted microbubbles have been developed to selectively adhere to specific and overexpressed p molecules in endothelial cells in some pathologic conditions.However,the law of

Rosiglitazone uppresses lipopolysaccharide-induced matrix metalloproteinase-2 activity in rat aortic endothelial cells via rasMEK1/2 signaling

ix metalloproteinase(MMPs) plays a key role in the pathogenesis of chronic inflammatory disease,such as atherosclerosis.Among MMPs,MMP-2 is regarded as a major proteinase in atherosclerotic plaque lesions.Peroxisome proliferator activated receptor-gamma(PPARg) ameliorates oxidative stress and the inflammatory response.The aim of the present study was to evaluate the effect of Rosiglitazone on Lipopolysaccharide(LPS)-induced MMP-2 activation as well as its possible mechanism.LPS-induced MMP-2 activity was inhibited by Rosiglitazone(PPARg agonist) in the rat aortic endothelial cells(RAEC).LPS-induced MMP-2 activation was diminished no matter exposure to NF-kB Activation Inhibitor II(JSH-23)or Ras inhibitor,farnesylthiosalicylic acid(FTS). Further study shows that LPS-induced activation of Phospho-Rho A and Phospho-MEKl/2 were significantly inhibited by Rosiglitazone.The activation of NF-kB p65 in the nuclear extract of cells was also significantly suppressed by Rosiglitazone, moreover,the expression of NF-κB p65 was partly activated by GW9662(PPARg antagonist).NF-kB DNA binding activity was also demolished by Rosiglitazone.In summary,our data showed that PPARg agonist,Rosiglitazone suppresses LPS-activated MMP-2 secretion via Ras-MEK1/2 signaling pathways and NF-kB activation.PPARg agonist and Ras-MEK1/2 pathway may be another potential therapeutic target for the disease induced by chronic inflammation.

Potentiation of NETs release is novel characteristic of TREM-1 activation and the pharmacological inhibition of TREM-1 could prevent from the deleterious consequences of NETs release in sepsis

During sepsis,neutrophil activation induces endothelial cell(EC)dysfunction partly through neutrophil extracellular trap(NET)release.The triggering receptor expressed on myeloid cell-1(TREM-1)is an orphan immune receptor that amplifies the inflammatory response mediated by Toll-like receptor-4(TLR4)engagement.Although the key role of TLR4 signaling in NETosis is known,the role of TREM-1 in this process has not yet been investigated.Here,we report that TREM-1 potentiates NET release by human and murine neutrophils and is a component of the NET structure.In contrast,pharmacologic inhibition or genetic ablation of TREM-1 decreased NETosis in vitro and during experimental septic shock in vivo.Moreover,isolated NETs were able to activate ECs and impair vascular reactivity,and these deleterious effects were dampened by TREM-1 inhibition.TREM-1 may,therefore,constitute a new therapeutic target to prevent NETosis and associated endothelial dysfunction.

Changes of activated circulating endothelial cells and survivin in patients with non-small cell lung cancer after antiangiogenesis therapy

Background Although antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells （aCECs） and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy. Methods Patients of non-small cell lung cancer （NSCLC） treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction （PCR）. Results After treatment, the amount of activated CECs decreased significantly in clinical benefit cases （P=0.021 in chemotherapy alone, P=0.001 in chemotherapy plus Endostar）, increased in disease progressive cases （P=0.015 in chemotherapy alone, but P=0.293 in chemotherapy with Endotatar）. After therapy, the expression of survivin mRNA decreased in clinical benefit cases （P=0.001） and increased in disease progressive cases （P=0.018）. A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy （P=0.001 and 0.021, respectively）, but only in the chemotherapy with Endostar group pre-therapy （P=0.030） rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression （TTP） （r=0.322, P=0.012）; a negative correlation was found between the amount of survivin mRNA in serum post-therapy and -l-I-P（r= -0.291, P=0.048）. Conclusions Activated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.

Functional nano-vector boost antiatherosclerosis efficacy of berberine in Apoe(-/-) mice

Atherosclerosis(AS)is the leading cause of heart attacks,stroke,and peripheral vascular disease.Berberine(BBR),a botanical medicine,has diversified anti-atherosclerotic effects but with poor absorption.The aim of this study was to develop an effective BBR-entrapped nano-system for treating AS in high-fat diet(HFD)-fed Apoe(-/-)mice,and also explore the possible underlying mechanisms involved.Three D-a-tocopherol polyethylene glycol(PEG)succinate(TPGS)analogues with different PEG chain lengths were synthesized to formulate BBR-entrapped micelles.HFD-fed Apoe(-/-)mice were administered with optimized formula(BBR,100 mg/kg/day)orally for 5 months.The artery plaque onset and related metabolic disorders were evaluated,and the underlying mechanisms were studied.Our data showed that,BT1500M increased BBR deposition in liver and adipose by 107.6%and 172.3%,respectively.In the Apoe(-/-)mice,BT1500M ameliorated HFD-induced hyperlipidemia and lipid accumulation in liver and adipose.BT1500M also suppressed HFD-induced chronic inflammation as evidenced by the reduced liver and adipose levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β);and decreased plasma level of TNF-α,IL-6,IL-1β,interferon-γ(IFN-γ),monocyte chemotactic protein(MCP),and macrophage inflammatory factor(MIP).The mechanism study showed that BT1500M changed Ampk and Nf-Kb gene expression,and interrupted a crosstalk process between adipocytes and macrophages.Further investigation proved that BT1500M decreased endothelial lesion and subsequent macrophage activation,cytokines release,as well as cholesteryl ester gathering in the aortic arch,resulting in ameliorated artery plaque build-up.Our results provide a practical strategy for treating AS using a BBR-entrapped nano-system.

TRIM65 E3 ligase targets VCAM-1 degradation to limit LPS-induced lung inflammation

Although the adhesion molecules-mediated leukocyte adherence and infiltration into tissues is an important step of inflammation,the post-translational regulation of these proteins on the endothelial cells is poorly understood.Here,we report that TRIM65,an ubiquitin E3 iigase of tripartite protein family,selectively targets vascular cell adhesion molecule 1(VCAM-1)and promotes its ubiq-uitination and degradation,by which it critically controls the duration and magnitude of sepsis-induced pulmonary inflammation.TRIM65 is constitutively expressed in human vascular endothelial cells.During TNFa-induced endothelial activation,the protein levels ofTRIM65 and VCAM-1 are inversely correlated.Expression of wild-type TRIM65,but not expression of aTRIM65 mutant that lacks E3 ubiquitin ligase function in endothelial cells,promotes VCAM-1 ubiquitination and degradation,whereas small interference RNA-mediated knockdown of TRIM65 attenuates VCAM-1 protein degradation.Further experiments show that TRIM65 directly interacts with VCAM-1 protein and directs its polyubiquitination,by which TRIM65 controls monocyte adherence and infiltration into tissues during inflammation.Importantly,TRIM65-deficient mice are more sensitive to lipopolysaccharide-induced death,due to sustained and severe pulmonary inflammation.Taken together,our studies suggest that TRIM65-mediated degradation of VCAM-1 represents a potential mechanism that controls the duration and magnitude of infiammation.