7 research outputs found
Fenofibrate protects endothelial cells against the harmful effects of TNF-alpha
Introduction: Fenofibrate exerts pleiotropic effects on endothelial cells (ECs) by, amongst others, increasing nitric oxide (NO) production. We aimed to investigate fenofi brate’s putative beneficial actions in healthy or TNF-alpha-induced dysfunctional ECs.Methods: Fenofi brate-induced pro-vasodilatory responses were assessed in aortic rings (50 - 125μM; 30min) with and without L-NMMA (100μM). Rat cardiac microvascular ECs were treated with fenofibrate (30 and 50μM; 1h). In the pre-treatment experiments, fenofibrate (50μM) was administered one hour before TNFalpha treatment (20ng/ml; 24h). NO-production (DAF-2/DA or Griess assay), mitochondrial ROS-production (MitoSox™), cell viability (propidium iodide staining), and changes in the expression/phosphorylation of critical endothelial proteins were measured by Western blotting.Results: Fenofibrate increased NO-production ˜2-fold in healthy ECs (p<0.05 vs. vehicle). A ˜23% pro-vasodilatory response was induced in aortic rings, which was reversed by L-NMMA (p<0.05 vs. fenofibrate). Fenofibrate pretreatment ameliorated TNF-alpha-induced endothelial dysfunction by reversing the loss of NO, improving oxidative stress, restoring cell viability and preventing caspase-3 activation. Protective effects were underpinned by ˜47% and ˜49% up-regulation of activated eNOS and AMP-kinase, respectively (p<0.05 vs. TNFalpha).Conclusions: Fenofibrate protects TNF-alpha-induced dysfunctional ECs via up-regulated eNOS-NO, reduced oxidative stress and improved cell viability. These novel findings warrant further investigations to explore the potential use of fenofibrate as an anti-endothelial dysfunction therapeutic agent
Cardiovascular risk and endothelial function in people living with HIV/AIDS: design of the multi-site, longitudinal EndoAfrica study in the Western Cape Province of South Africa
CITATION: Strijdom, H., et al. 2017. Cardiovascular risk and endothelial function in people living with HIV/AIDS: design of the multi-site, longitudinal EndoAfrica study in the Western Cape Province of South Africa. BMC Infectious Diseases, 17:41, doi:10.1186/s12879-016-2158-y.The original publication is available at http://bmcinfectdis.biomedcentral.comBackground: There is growing evidence of an interaction between HIV-infection, anti-retroviral therapy (ART) and
cardiovascular diseases (CVD). Epidemiological studies in Europe and North America have been observing a shift
towards an increased incidence of coronary heart disease and acute myocardial infarctions in HIV-infected populations
compared to the general population even after adjusting for traditional cardiovascular risk factors. Despite South Africa
(and sub-Saharan Africa, SSA) being regarded as the epicentre of the global HIV epidemic, very little is known about
the prevalence of cardiovascular risk factors and precursors of vascular disease in HIV-infected populations in this
region. The knowledge gap is further widened by the paucity of data from prospective studies. We present the
rationale, objectives and key methodological features of the EndoAfrica study, which aims to determine whether HIVinfection
and ART are associated with altered cardiovascular risk and changes in vascular endothelial structure and
function in adults living in the Western Cape Province of South Africa.
Methods: In this longitudinal study, comprehensive cardiovascular assessments of HIV-negative and HIV-positive (with
and without ART) study participants are performed by clinical and biochemical screening for traditional cardiovascular
risk factors and biomarkers of CVD. Vascular and endothelial function is determined by brachial artery flow-mediated
dilatation (FMD), carotid-intima-thickness (IMT) measurements and quantitative retinal blood vessel analyses,
complemented by vascular endothelial biomarker assays. Finally, we aim to statistically determine whether HIVinfection
and/or ART are associated with increased cardiovascular risk and vascular endothelial dysfunction, and
determine whether there is progression/regression in these endpoints 18 months after the baseline assessments.
Discussion: The EndoAfrica study provides a unique opportunity to recruit a cohort of HIV-infected patients and HIVnegative
controls who will be comprehensively and longitudinally assessed for cardiovascular risk and disease profile
with vascular endothelial function as a potentially important intermediate cardiovascular phenotype. To our
knowledge, it is the first time that such a systematic study has been established in the context of SSA and South Africa.http://bmcinfectdis.biomedcentral.com/articles/10.1186/s12879-016-2158-yPublisher's versio
Investigating the cholesterol-independent (pleiotropic) effects of selected hypolipidaemic agents in functional and dysfunctional endothelial cells
Thesis (DScMedSc)--Stellenbosch University, 2015.ENGLISH ABSTRACT: Vascular endothelium forms the first line of defence against harmful stimuli in the circulation.
Endothelial dysfunction is a valuable predictor of cardiovascular disease and therapies aimed at
improving endothelial function are therefore needed. The anti-dyslipidaemic agents, simvastatin
and fenofibrate, are known for their beneficial effects on lipid parameters, however additional
pleiotropic effects have been shown for both. These include improved endothelial function due
to increased levels of nitric oxide (NO), as well as anti-oxidant and anti-inflammatory actions. NO
is produced by the enzyme, nitric oxide synthase (NOS), which exists in the endothelial NOS
(eNOS), inducible NOS (iNOS) and neuronal NOS (nNOS) isoforms. Most studies investigating the
endothelial effects of simvastatin and fenofibrate are performed on macrovascular-derived
endothelial cells, and there is a lack of data on endothelial cells (ECs) from the microcirculation,
particularly the cardiac microvessels.
This dissertation aimed to investigate and elucidate mechanisms underlying the pleiotropic
effects of simvastatin and fenofibrate on ECs and vascular tissue using in vitro, ex vivo and in vivo
experimental models. In vitro investigations included flow cytometry-based intracellular
measurements of NO, as well as different types of reactive oxygen species (ROS) and cell viability
parameters. Signalling pathways involved with these changes were measured by western blot
analyses of the expression and phosphorylation of critical proteins involved in vascular function.
Results on cardiac microvascular ECs (CMECs) demonstrated that fenofibrate (50 μM) exerted a
potent, increasing effect on NO production after short periods (1 and 4 hour treatments), but
after 24 hours the effects were less robust. Exhaustive investigations suggested that the NOincreasing
effects of fenofibrate in baseline CMECs were NOS-independent, a novel finding as far
as we are aware. Fenofibrate’s ability to protect ECs against injury was demonstrated when
CMECs incubated with the pro-inflammatory cytokine, TNF-α, were pre-treated with fenofibrate,
resulting in increased NO and improved cell viability parameters. Simvastatin (1 μM) increased
NO to a lesser extent in baseline CMECs, and resulted in increased apoptosis and necrosis.
Following the cell studies, their effects on vascular reactivity was measured by aortic ring
isometric tension studies. The effects of acutely administered fenofibrate to pre-contracted
aortic rings were investigated, and results showed a modest, but significant NOS-dependent
vasodilatory response. Next, an in vivo model of Wistar rats treated with simvastatin (0.5
mg/kg/day) and fenofibrate (100 mg/kg/day) for 6 weeks was established. Data showed that
neither drug was able to improve aortic ring contraction and dilation above baseline values. Both
drug treatments increased iNOS expression, which is usually associated with harmful actions.
However, in our hands, increased iNOS expression was associated with a beneficial anticontractile
response in the simvastatin-treated animals. Fenofibrate treatment increased NO
bioavailability in the blood of these animals.
In conclusion, fenofibrate showed endothelio-protective pleiotropic effects with regards to NO
production after short treatment periods in CMECs. These effects were mediated via a NOSindependent
mechanism, a novel finding. Fenofibrate pre-treatment was also protective against
the harmful effects of TNF-α. Simvastatin did not show pronounced pleiotropic effects in vitro
or in vivo on endothelial function.AFRIKAANSE OPSOMMING: Die vaskulêre endoteellaag is die eerste linie van verdediging teen skadelike stimuli in die
bloedsirkulasie. Endoteeldisfunksie is ‘n waardevolle voorspeller van kardiovaskulêre siektes en
enige terapeutiese behandeling wat kan bydra tot verbeterde endoteelfunksie is belangrik.
Simvastatien en fenofibraat word as anti-dislipidemiese middels voorgeskryf en hoewel hulle
primêr gebruik word om cholesterolvlakke te verbeter, toon hulle ook pleiotropiese (cholesterolonafhanklike)
eienskappe. Dit sluit in bevordering van endoteelfunksie (via verhoogde
stikstofoksied (NO) produksie), asook anti-oksidant en anti-inflammatoriese effekte. NO word
vervaardig deur die ensiem, stikstofoksiedsintase (NOS) wat voorkom in drie isovorme: endoteelafgeleide
NOS (eNOS), induseerbare NOS (iNOS) en neuronale NOS (nNOS). Die meerderheid
studies wat pleiotropiese effekte van simvastatien en fenofibraat ondersoek, gebruik
endoteelselle van makrovaskulêre bloedvate, wat beteken daar is ‘n tekort aan data aangaande
endoteelselle vanaf mikrovaskulêre vate, veral kardiale mikrovaskulêre vate (CMECs).
Hierdie proefskrif het dit ten doel gehad om meganismes betrokke by die pleiotropiese effekte
van simvastatien en fenofibraat te ondersoek deur van in vitro, ex vivo en in vivo modelle gebruik
te maak. Die in vitro ondersoeke het gefokus op vloeisitometrie-gebaseerde metings van
intrasellulêre NO, reaktiewe suurstof-radikale (ROS) en sellewensvatbaarheid. Seintransduksie
paaie betrokke by hierdie veranderinge was bepaal deur proteienuitdrukking en -fosforilasie
vlakke te meet van belangrike proteïene, met behulp van die Western-blot tegniek.
Resultate van die CMEC eksperimente het getoon dat fenofibraat (50 μM) ‘n kragtige en
verhogende effek op NO produksie uitgeoefen het na kort behandelingstye (1 en 4 ure), maar na
24 uur was hierdie effek minder uitgesproke. Uitvoerige ondersoeke het getoon dat fenofibraat
se basislyn effekte op CMECs deur NOS-onafhanklike meganismes teweeggebring is, en sover ons
kennis strek, is dit ‘n nuwe bevinding. Fenofibraat se endoteel-beskermende effekte kon ook
aangetoon word deur CMECs vir een uur te behandel voor byvoeging van die pro-inflammatories
sitokien, tumor nekrose faktor alpha (TNF-α), wat gelei het tot verhoogde NO vlakke en
verbeterde seloorlewing. Simvastatien (1 μM) het tot ‘n mindere mate NO produksie verhoog in
CMECs, tesame met pro-apoptotiese en -nekrotiese effekte.
Vervolgens was die effekte op vaskulêre reaktiwiteit geëvalueer d.m.v. isometriese
spanningsondersoeke. Akute effekte van fenofibraat is gemeet deur byvoeging daarvan tot ‘n
vooraf saamgetrekte aorta-ring, wat tot matige, maar beduidende NOS-afhanklike verslapping
gelei het. Hierna is ‘n in vivo model opgestel deur Wistar rotte vir ses weke met 0.5 mg/kg/dag
simvastatien of 100 mg/kg/dag fenofibraat te behandel. Resultate toon dat geen van die
behandelings basislyn kontraksie of verslapping van aorta ringe kon verbeter nie. Beide
behandelings het tot verhoogde iNOS uitdrukking gelei, wat gewoonlik met nadelige effekte
geassosieer word, maar in ons studies was dit met voordelige, anti-kontraktiele effekte in aortaringe
van simvastatien-behandelde rotte geassosieer. Fenofibraat behandeling het die NObiobeskikbaarheid
in die rotte se bloed verhoog.
Ten slotte, fenofibraat het met endoteel-beskermende, pleiotropiese effekte op endoteelselle
gepaard gegaan, veral t.o.v. NO-produksie na akute middeltoediening in die CMECs. Die
meganisme was ‘n NOS-onafkanklike proses, wat ‘n nuwe bevinding is. Fenofibraat prebehandeling
het teen die skadelike effekte van TNF-α beskerm. Geen uitgesproke pleiotropiese
effekte is in vitro of in vivo gevind met simvastatien behandeling nie
Fenofibrate protects endothelial cells against the harmful effects of TNF-alpha
CITATION: Westcott, C. et al. 2017. Fenofibrate protects endothelial cells against the harmful effects of TNF-alpha. SA Heart, 14(1):22-34, doi:10.24170/14-1-1865.The original publication is available at http://www.journals.ac.za/index.php/SAHJIntroduction: Fenofibrate exerts pleiotropic effects on endothelial cells (ECs) by, amongst others, increasing nitric oxide (NO) production. We aimed to investigate fenofi brate’s putative beneficial actions in healthy or TNF-alpha-induced dysfunctional ECs.
Methods: Fenofi brate-induced pro-vasodilatory responses were assessed in aortic rings (50 - 125μM; 30min) with and without L-NMMA (100μM). Rat cardiac microvascular ECs were treated with fenofibrate (30 and 50μM; 1h). In the pre-treatment experiments, fenofibrate (50μM) was administered one hour before TNFalpha treatment (20ng/ml; 24h). NO-production (DAF-2/DA or Griess assay), mitochondrial ROS-production (MitoSox™), cell viability (propidium iodide staining), and changes in the expression/phosphorylation of critical endothelial proteins were measured by Western blotting.
Results: Fenofibrate increased NO-production ˜2-fold in healthy ECs (p<0.05 vs. vehicle). A ˜23% pro-vasodilatory response was induced in aortic rings, which was reversed by L-NMMA (p<0.05 vs. fenofibrate). Fenofibrate pretreatment ameliorated TNF-alpha-induced endothelial dysfunction by reversing the loss of NO, improving oxidative stress, restoring cell viability and preventing caspase-3 activation. Protective effects were underpinned by ˜47% and ˜49% up-regulation of activated eNOS and AMP-kinase, respectively (p<0.05 vs. TNFalpha).
Conclusions: Fenofibrate protects TNF-alpha-induced dysfunctional ECs via up-regulated eNOS-NO, reduced oxidative stress and improved cell viability. These novel findings warrant further investigations to explore the potential use of fenofibrate as an anti-endothelial dysfunction therapeutic agent.http://www.journals.ac.za/index.php/SAHJ/article/view/1865Publisher's versio
Vascular health assessment with flow-mediated dilatation and retinal image analysis : a pilot study in an adult population from Cape Town
AIM: Flow-mediated dilatation (FMD) and retinal vascular analysis (RVA) may assist in predicting cardiovascular disease (CVD) but are poorly characterised in South Africa. We recorded baseline FMD and retinal vascular widths in healthy participants, investigated associations with cardiovascular risk factors. METHODS: Endothelial function (measured with FMD), microvascular structure (evaluated via fundus image analysis) and major CVD risk factors were assessed in 66 participants from Cape Town. RESULTS: Median FMD% was 9.6%, with higher values in females. Mean retinal arteriolar and venular widths were ~156 and ~250 μm, respectively. FMD was not associated with CVD risk factors. Hypertension was associated with narrower retinal arterioles and venules. CONCLUSIONS: We report novel baseline FMD data in healthy South African adults from the Western Cape, and show that retinal microvascular calibres are associated with blood pressure. Our baseline FMD and RVA data could serve as a reference for future studies in South Africa
Cardiovascular risk and endothelial function in people living with HIV/AIDS: design of the multi-site, longitudinal EndoAfrica study in the Western Cape Province of South Africa
There is growing evidence of an interaction between HIV-infection, anti-retroviral therapy (ART) and cardiovascular diseases (CVD). Epidemiological studies in Europe and North America have been observing a shift towards an increased incidence of coronary heart disease and acute myocardial infarctions in HIV-infected populations compared to the general population even after adjusting for traditional cardiovascular risk factors. Despite South Africa (and sub-Saharan Africa, SSA) being regarded as the epicentre of the global HIV epidemic, very little is known about the prevalence of cardiovascular risk factors and precursors of vascular disease in HIV-infected populations in this region. The knowledge gap is further widened by the paucity of data from prospective studies. We present the rationale, objectives and key methodological features of the EndoAfrica study, which aims to determine whether HIV-infection and ART are associated with altered cardiovascular risk and changes in vascular endothelial structure and function in adults living in the Western Cape Province of South Africa.status: publishe