siRNA knocks down Hsp27 and increases angiotensin II-Induced phosphorylated NF-κB p65 levels in aortic smooth muscle cells

Voegeli, Tracy S.; Currie, R. William

doi:10.1007/s00011-009-8166-2

Cited by 15 publications

(10 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Levels of phosphorylated NF-κB p65, IκBα, and β-actin were analyzed by western blotting [29]-[31]. The 3D keratinocytes were lysed in lysis buffer (10 mM Tris-HCl, pH 7.5, 50 mM NaCl, 10 mM NaF, 2 mM Na 3 VO 4 , 1 mM DFP, 1% Triton X-100, with 1/20 v/v Complete) and sonicated for 10 s with 30% output using an ultrasonic sonifier (model 250; Branson Ultrasonics; Danbury, CT, USA).…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas-Derived Ceramidase Induces Production of Inflammatory Mediators from Human Keratinocytes via Sphingosine-1-Phosphate

et al. 2014

View full text Add to dashboard Cite

Ceramide is important for water retention and permeability barrier functions in the stratum corneum, and plays a key role in the pathogenesis of atopic dermatitis (AD). A Pseudomonas aeruginosa-derived neutral ceramidase (PaCDase) isolated from a patient with AD was shown to effectively degrade ceramide in the presence of Staphylococcus aureus-derived lipids or neutral detergents. However, the effect of ceramide metabolites on the functions of differentiating keratinocytes is poorly understood. We found that the ceramide metabolite sphingosine-1-phosphate (S1P) stimulated the production of inflammatory mediators such as TNF-α and IL-8 from three-dimensionally cultured human primary keratinocytes (termed “3D keratinocytes”), which form a stratum corneum. PaCDase alone did not affect TNF-α gene expression in 3D keratinocytes. In the presence of the detergent Triton X-100, which damages stratum corneum structure, PaCDase, but not heat-inactivated PaCDase or PaCDase-inactive mutant, induced the production of TNF-α, endothelin-1, and IL-8, indicating that this production was dependent on ceramidase activity. Among various ceramide metabolites, sphingosine and S1P enhanced the gene expression of TNF-α, endothelin-1, and IL-8. The PaCDase-enhanced expression of these genes was inhibited by a sphingosine kinase inhibitor and by an S1P receptor antagonist VPC 23019. The TNF-α-binding antibody infliximab suppressed the PaCDase-induced upregulation of IL-8, but not TNF-α, mRNA. PaCDase induced NF-κB p65 phosphorylation. The NF-κB inhibitor curcumin significantly inhibited PaCDase-induced expression of IL-8 and endothelin-1. VPC 23019 and infliximab inhibited PaCDase-induced NF-κB p65 phosphorylation and reduction in the protein level of the NF-κB inhibitor IκBα. Collectively, these findings suggest that (i) 3D keratinocytes produce S1P from sphingosine, which is produced through the hydrolysis of ceramide by PaCDase, (ii) S1P induces the production of TNF-α via S1P receptors, and (iii) released TNF-α stimulates the production of inflammatory mediators such as IL-8.

show abstract

Section: Methodsmentioning

confidence: 99%

Pseudomonas-Derived Ceramidase Induces Production of Inflammatory Mediators from Human Keratinocytes via Sphingosine-1-Phosphate

et al. 2014

View full text Add to dashboard Cite

show abstract

“…For instance, compound echinacoside (mol41) targets the VEGF pathway through protein HSPB1(Supplementary Figure 1 ). Fortunately, studies show that HSPB1 has significant cytoprotective properties in several models of neurological disease in vivo 31 , 32 or in vitro 33 and it may play a role in anti-inflammatory effect by regulating the nuclear factor-κB (NF-κB) signaling pathway 34 , 35 . In addition, a study also demonstrate that R-Ras could regulate angiogenic activities of endothelial cells in part via inhibition of the p38 mitogen-activated protein kinase (p38 MAPK)-HSPB1 axis of the VEGF signaling pathway 36 .…”

Section: Resultsmentioning

confidence: 99%

In silico-based screen synergistic drug combinations from herb medicines: a case using Cistanche tubulosa

Liu

Zhu

Xue

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Neuroinflammation is characterized by the elaborated inflammatory response repertoire of central nervous system tissue. The limitations of the current treatments for neuroinflammation are well-known side effects in the clinical trials of monotherapy. Drug combination therapies are promising strategies to overcome the compensatory mechanisms and off-target effects. However, discovery of synergistic drug combinations from herb medicines is rare. Encouraged by the successfully applied cases we move on to investigate the effective drug combinations based on system pharmacology among compounds from Cistanche tubulosa (SCHENK) R. WIGHT. Firstly, 63 potential bioactive compounds, the related 133 direct and indirect targets are screened out by Drug-likeness evaluation combined with drug targeting process. Secondly, Compound-Target network is built to acquire the data set for predicting drug combinations. We list the top 10 drug combinations which are employed by the algorithm Probability Ensemble Approach (PEA), and Compound-Target-Pathway network is then constructed by the 12 compounds of the combinations, targets, and pathways to unearth the corresponding pharmacological actions. Finally, an integrating pathway approach is developed to elucidate the therapeutic effects of the herb in different pathological features-relevant biological processes. Overall, the method may provide a productive avenue for developing drug combination therapeutics.

show abstract

“…The cells were placed in a 5% CO 2 incubator at 37°C and 8 hrs after siRNA administration the media was replaced with fresh media without antibiotics. After 48 hrs total elapsed time, the cells were treated with FL3 (100 nM) for 10 h and then were treated with doxorubicin for an additional 14 h. Hsp27 siRNA (Cat #54502) and a negative control siRNA (Cat #4611) were purchased from Ambion [17]. The working concentration of siRNA in cell experiments was 30 nM.…”

Section: Methodsmentioning

confidence: 99%

Flavaglines Alleviate Doxorubicin Cardiotoxicity: Implication of Hsp27

et al. 2011

View full text Add to dashboard Cite

BackgroundDespite its effectiveness in the treatment of various cancers, the use of doxorubicin is limited by a potentially fatal cardiomyopathy. Prevention of this cardiotoxicity remains a critical issue in clinical oncology. We hypothesized that flavaglines, a family of natural compounds that display potent neuroprotective effects, may also alleviate doxorubicin-induced cardiotoxicity.Methodology/Principal FindingsOur in vitro data established that a pretreatment with flavaglines significantly increased viability of doxorubicin-injured H9c2 cardiomyocytes as demonstrated by annexin V, TUNEL and active caspase-3 assays. We demonstrated also that phosphorylation of the small heat shock protein Hsp27 is involved in the mechanism by which flavaglines display their cardioprotective effect. Furthermore, knocking-down Hsp27 in H9c2 cardiomyocytes completely reversed this cardioprotection. Administration of our lead compound (FL3) to mice attenuated cardiomyocyte apoptosis and cardiac fibrosis, as reflected by a 50% decrease of mortality.Conclusions/SignificanceThese results suggest a prophylactic potential of flavaglines to prevent doxorubicin-induced cardiac toxicity.

show abstract

siRNA knocks down Hsp27 and increases angiotensin II-Induced phosphorylated NF-κB p65 levels in aortic smooth muscle cells

Cited by 15 publications

References 18 publications

Pseudomonas-Derived Ceramidase Induces Production of Inflammatory Mediators from Human Keratinocytes via Sphingosine-1-Phosphate

Pseudomonas-Derived Ceramidase Induces Production of Inflammatory Mediators from Human Keratinocytes via Sphingosine-1-Phosphate

In silico-based screen synergistic drug combinations from herb medicines: a case using Cistanche tubulosa

Flavaglines Alleviate Doxorubicin Cardiotoxicity: Implication of Hsp27

Contact Info

Product

Resources

About