Correction of vascular hypercontractility in spontaneously hypertensive rats using shRNAs-induced delta protein kinase C gene silencing

Novokhatska, Tetiana; Tishkin, Sergey; Dosenko, Victor; Boldyriev, Oleksiy I.; Ivanova, Irina; Strielkov, Ievgen; Soloviev, Anatoly I.

doi:10.1016/j.ejphar.2013.08.003

Cited by 19 publications

(21 citation statements)

References 32 publications

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“…Our data show that bryostatin-1 decreased levels of PKCα when given after blast. A separate isozyme, PKCδ, mediates vasoconstriction, but its role in tight junction disruption is poorly understood [64]. Our data show that bryostatin-1 had a negligible effect on PKCδ levels.…”

Section: Discussionmentioning

confidence: 72%

Bryostatin-1 Restores Blood Brain Barrier Integrity following Blast-Induced Traumatic Brain Injury

et al. 2014

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Recent wars in Iraq and Afghanistan have accounted for an estimated 270,000 blast exposures among military personnel. Blast traumatic brain injury (TBI) is the ‘signature injury’ of modern warfare. Blood brain barrier (BBB) disruption following blast TBI can lead to long-term and diffuse neuroinflammation. In this study, we investigate for the first time the role of bryostatin-1, a specific protein kinase C (PKC) modulator, in ameliorating BBB breakdown. Thirty seven Sprague–Dawley rats were used for this study. We utilized a clinically relevant and validated blast model to expose animals to moderate blast exposure. Groups included: control, single blast exposure, and single blast exposure + bryostatin-1. Bryostatin-1 was administered i.p. 2.5 mg/kg after blast exposure. Evan’s blue, immunohistochemistry, and western blot analysis were performed to assess injury. Evan’s blue binds to albumin and is a marker for BBB disruption. The single blast exposure caused an increase in permeability compared to control (t=4.808, p<0.05), and a reduction back toward control levels when bryostatin-1 was administered (t=5.113, p<0.01). Three important PKC isozymes, PKCα, PKCδ, and PKCε, were co-localized primarily with endothelial cells but not astrocytes. Bryostatin-1 administration reduced toxic PKCα levels back toward control levels (t=4.559, p<0.01) and increased the neuroprotective isozyme PKCε (t=6.102, p<0.01). Bryostatin-1 caused a significant increase in the tight junction proteins VE-cadherin, ZO-1, and occludin through modulation of PKC activity. Bryostatin-1 ultimately decreased BBB breakdown potentially due to modulation of PKC isozymes. Future work will examine the role of bryostatin-1 in preventing chronic neurodegeneration following repetitive neurotrauma.

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Section: Discussionmentioning

confidence: 72%

Bryostatin-1 Restores Blood Brain Barrier Integrity following Blast-Induced Traumatic Brain Injury

et al. 2014

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“…Recent studies showed that PKC is a key pathological factor in hypertension. 29;46-48 PKC mediates its physiological effects mainly through 13 isoforms, divided into 3 major groups: 1) classical PKC (cPKC) isoforms (α, βI, βII, and γ) requiring both Ca 2+ and diacylglycerol (DAG) for activation, 2) novel PKC (nPKC) isoforms (δ, ε, η, and θ) requiring only DAG for activation, and 3) atypical PKC isoforms (λ, μ, and ζ) that are activated by binding phosphatidylserine but not DAG or Ca 2+ . 49 The α, β, δ, ε and ζ isoenzymes of PKC have been detected in vascular smooth muscles.…”

Section: Discussionmentioning

confidence: 99%

“…29-32 For example, PKC-mediated contractile responses are enhanced in aortas of SHR but not Wistar-Kyoto rats (WKY); 33;34 hypertensive models, including the SHR 35;36 and DOCA-salt hypertensive rats have increased vascular expression/activity of PKC; 37 perfusion of hindlimb of SHR and WKY rats with the PKC activator, phorbol 12,13-dibutyrate (PDBu) caused prolonged vasoconstriction and increased perfusion pressure; 38 and PKC gene silencing normalized arterial BP in SHR. 29 These studies suggest that increased PKC expression and function may be a key molecule contributing to the development of hypertension. However, the molecular basis of activation of PKCs and the mechanisms that control the expression of PKC isozymes, especially knowledge regarding the functional elements in the PKC gene promoter or the nature of the factors that control PKCδ gene transcription is not well characterized.…”

Section: Introductionmentioning

confidence: 99%

Prenatal Testosterone Exposure Induces Hypertension in Adult Females via Androgen Receptor–Dependent Protein Kinase Cδ–Mediated Mechanism

et al. 2015

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Prenatal exposure to excess testosterone induces hyperandrogenism in adult females and predisposes them to hypertension. We tested whether androgens induce hypertension through transcriptional regulation and signaling of protein kinase C (PKC) in the mesenteric arteries. Pregnant Sprague-Dawley rats were injected with vehicle or testosterone propionate (0.5mg/kg/day from gestation day 15–19, subcutaneously) and their 6-month-old adult female offspring were examined. Plasma testosterone levels (0.84±0.04 vs 0.42±0.09 ng/ml) and blood pressures (111.6± 1.3 vs 104.5 ± 2.4 mmHg) were significantly higher in prenatal testosterone-exposed rats compared to controls. This was accompanied with enhanced expression of PKCδ mRNA (1.5-fold) and protein (1.7-fold) in the mesenteric arteries of prenatal testosterone-exposed rats. In addition, mesenteric artery contractile responses to PKC activator—phorbol-12,13-dibutyrate—was significantly greater in prenatal testosterone-exposed rats. Treatment with androgen receptor antagonist flutamide (10 mg/kg, subcutaneously, twice-daily for 10 days) significantly attenuated hypertension, PKCδ expression, and the exaggerated vasoconstriction in prenatal testosterone-exposed rats. In vitro exposure of testosterone to cultured mesenteric artery smooth muscle cells dose-dependently upregulated PKCδ expression. Analysis of PKCδ gene revealed a putative androgen responsive element in the promoter upstream to the transcription start site and an enhancer element in intron-1. Chromatin-immunoprecipitation assays showed that androgen receptors bind to these elements in response to testosterone stimulation. Furthermore, luciferase reporter assays showed that the enhancer element is highly responsive to androgens and treatment with flutamide reverses reporter activity. Our studies identified a novel androgen-mediated mechanism for the control of PKCδ expression via transcriptional regulation that controls vasoconstriction and blood pressure.

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“…Large conductance Ca 2+ -activated K + channels (BK Ca ) are the predominant K + channels in VSMCs (Nelson and Quayle, 1995; Ghatta et al, 2006). PKC activators such as PDBu inhibit BK Ca leading to increases in vascular tone in both physiological and pathophysiological conditions (Taguchi et al, 2000; Barman et al, 2004; Kizub et al, 2010; Novokhatska et al, 2013), and in various vascular beds including pulmonary (Barman et al, 2004), coronary (Minami et al, 1993), cerebral (Lange et al, 1997), and uterine vessels (Hu et al, 2011). PKC activators inhibit BK Ca by phosphorylation of the channel protein and decreasing its sensitivity to activation by cGMP-dependent protein kinase (Crozatier, 2006; Ledoux et al, 2006).…”

Section: Vascular Effects Of Pkcmentioning

confidence: 99%

Protein Kinase C as Regulator of Vascular Smooth Muscle Function and Potential Target in Vascular Disorders

Ringvold

Khalil

2017

Advances in Pharmacology

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Vascular smooth muscle (VSM) plays an important role in maintaining vascular tone. In addition to Ca2+-dependent myosin light chain (MLC) phosphorylation, protein kinase C (PKC) is a major regulator of VSM function. PKC is a family of conventional Ca2+-dependent α, β, and γ, novel Ca2+-independent δ, ε, θ, and η, and atypical ξ, and ι/λ isoforms. Inactive PKC is mainly cytosolic, and upon activation it undergoes phosphorylation, maturation and translocation to the surface membrane, the nucleus, endoplasmic reticulum, and other cell organelles; a process facilitated by scaffold proteins such as RACKs. Activated PKC phosphorylates different substrates including ion channels, pumps and nuclear proteins. PKC also phosphorylates CPI-17 leading to inhibition of MLC phosphatase, increased MLC phosphorylation and enhanced VSM contraction. PKC could also initiate a cascade of protein kinases leading to phosphorylation of the actin-binding proteins calponin and caldesmon, increased actin-myosin interaction and VSM contraction. Increased PKC activity has been associated with vascular disorders including ischemia-reperfusion injury, coronary artery disease, hypertension, and diabetic vasculopathy. PKC inhibitors could test the role of PKC in different systems, and could reduce PKC hyperactivity in vascular disorders. First generation PKC inhibitors such as staurosporine and chelerythrine are not very specific. Isoform-specific PKC inhibitors such as ruboxistaurin have been tested in clinical trials. Target-delivery of PKC pseudosubstrate inhibitory peptides and PKC siRNA may be useful in localized vascular disease. Further studies of PKC and its role in VSM should help design isoform-specific PKC modulators that are experimentally potent and clinically safe to target PKC in vascular disease.

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Correction of vascular hypercontractility in spontaneously hypertensive rats using shRNAs-induced delta protein kinase C gene silencing

Cited by 19 publications

References 32 publications

Bryostatin-1 Restores Blood Brain Barrier Integrity following Blast-Induced Traumatic Brain Injury

Bryostatin-1 Restores Blood Brain Barrier Integrity following Blast-Induced Traumatic Brain Injury

Prenatal Testosterone Exposure Induces Hypertension in Adult Females via Androgen Receptor–Dependent Protein Kinase Cδ–Mediated Mechanism

Protein Kinase C as Regulator of Vascular Smooth Muscle Function and Potential Target in Vascular Disorders

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