Department of Vascular Pathophysiology

Department of Vascular Patophysiology is focused on complete investigation of functional, structural and molecular changes of cardiovascular system in normal and pathological conditions.

Functional studies are realized at the level of in vitro and in vivo experiments. Our research is aimed on investigation of vasoactive properties of isolated conduit and resistance arteries using the measurement of changes in isometric tension (in vitro). Specifically, we are focused on humoral and nerve (activity of perivascular nerves) regulation At the level of in vivo experiments we evaluate the integrated response of intact cardiovascular system – invasive measurement of blood pressure, heart frequention, pulse wave velocity and the determination of its characteristic points.

Structural changes are studied using light microscopy and immunohistochemistry. In light microscopy the geometry of the arteries (inner diameter, wall thickness and cross sectional area) is evaluated. The changes in the structure of arterial wall and volume densities of individual parts of arterial wall are investigated on ultrathin sections.

At the molecular level, changes of the redox regulation and signalization in different tissues, the expression of genes and proteins and activity of enzymes participating on blood pressure regulation and cardiovascular disorders are observed in normotensive and hypertensive animals and on the models of cultured dells. The used molecular and biochemical methods include UV-VIS-spectrophotometry, the measurement of radiolabelled isotopes, chemiluminiscent method for the measurement of superoxide, methods of direct fluorescence and immunofluorescence using fluorescence microscope, WB analysis and real-time (quantitative PCR).

Studies on animal model (rat) are correlated with the following of vasoactive properties and molecular changes of perivascular adipose tissue of intrarenal arteries isolated after nephrectomy of patients with carcinoma (cooperation with Urological clinic of Acad. L. Derer Hospital).

 

Head:

RNDr. Soňa Čačányiová, PhD.

tel.: +421 2 3229 6014

e-mail: sona.cacanyiova@savba.sk

 

Researchers:

Mgr. Andrea Berényiová, PhD.

RNDr. Magdaléna Drobná, PhD.

Mgr. Miroslava Majzúnová, PhD.

Doc. MUDr. Jozef Török, CSc.

Mgr. Zuzana Poljak Valášková, PhD.

Mgr. Anna Zemančíková, PhD.

 

PhD students:

Mgr. Samuel Golas

 

Laboratory technician:

RNDr. Eva Labudová

 

Undergraduate students:

Lukáš Prachár (Faculty of Natural Sciences, Comenius University)

Romana Ružinská (Faculty of Natural Sciences, Comenius University)

Dominika Krištofová (Faculty of Natural Sciences, Comenius University)

Ivana Kušnierová (Faculty of Chemical and Food Technology )

 


The most important publications in last 5 years

WU, Kay L.H. – CHAO, Yung-Mei – TSAY, Shiow-Jen – CHEN, Chen Hsiu – CHAN, Samuel H.H. – DOVINOVÁ, Ima – CHAN JULIE, Y.H. Role of nitric oxide synthase uncoupling at rostral ventrolateral medulla in redox-sensitive hypertension associated with metabolic syndrome. In Hypertension, 2014, vol. 64, no. 4, p.815-824. (7.632 – IF2013).

BERÉNYIOVÁ, Andrea – GRMAN, Marián – MIJUSKOVIC, A. – STAŠKO, Andrej – MIŠÁK, Anton – NAGY, Peter – ONDRIAŠOVÁ, Elena – ČAČÁNYIOVÁ, Soňa – BREZOVÁ, Vlasta – FEELISCH, Martin – ONDRIAŠ, Karol. The reaction products of sulfide and S-nitrosoglutathione are potent vasorelaxants. In Nitric Oxide : Biology and Chemistry, 2015, vol. 46, p. 123-130. (3.521 – IF2014).

TÖRÖK, Jozef – ZEMANČÍKOVÁ, Anna – KOCIANOVÁ, Z. Interaction of perivascular adipose tissue and sympathetic nerves in arteries from normotensive and hypertensive rats. In Physiological Research, 2016, vol. 65, suppl. 3, p. S391-S399. (1.618 – IF2015).

ČAČÁNYIOVÁ, Soňa – BERÉNYIOVÁ, Andrea – BALIŠ, Peter – KRISTEK, František – GRMAN, Marián – ONDRIAŠ, Karol – BREZA, J. – BREZA, J. Jr. Nitroso-sulfide coupled signaling triggers specific vasoactive effects in the intrarenal arteries of patients with arterial hypertension. In Journal of Physiology and Pharmacology: formerly Acta Physiologica Polonica, 2017, vol. 68, no. 4, p. 527-538. (2.883 – IF2016, Q1).

MAJZÚNOVÁ, Miroslava – PAKANOVÁ, Zuzana – KVASNIČKA, Peter – BALIŠ, Peter – ČAČÁNYIOVÁ, Soňa – DOVINOVÁ, Ima. Age-dependent redox status in the brain stem of NO-deficient hypertensive rats. In Journal of Biomedical Science, 2017, vol. 24, article 72, 14 p. (2.799 – IF2016, Q1)

 


Current projects

APVV

New regulatory effects of nitric oxide and their role in the development of essential hypertension

Principal investigator: RNDr. Soňa Čačányiová, PhD.

Lasting of the project: 1.7. 2016 – 30.6. 2020

Registration number of the project: APVV-15-0565

Anotation:

High blood pressure is a main risk factor in sustained increased morbidity and mortality of humans suffering cardiovascular diseases. Understanding of causes leading to hypertension enable to reveal new preventive and therapeutic decisions. A new regulatory system involved in vessel tree regulation seems to be neuronal NOsynthase (nNOS) and its interactions with other regulatory systems. On the level of the kidney nNOS signal pathway interferes with renin-angiotensin system (RAS) and sulfide signalization (H2S), and the interactions among them are the unexplored area of regulatory mechanisms. nNOS in macula densa stimulate renin syntesis and via it influences RAS and sympathetic nerve system, on the other hand, H2S inhibits renin synthesis. Moreover, the regulatory pathways of nNOS and also endothelial eNOS could interact with endogenous NOS inhibitor, asymmetric dimetylarginine ADMA, on local as well as systemic level. The aim of the project is to study the effect of interactions of NO/nNOS/eNOS signalization with mentioned regulatory pathways (RAS, H2S, ADMA) on cardiovascular system and to find out their role in the specificity of nNOS and/or eNOS action in the conditions of essential hypertension. The availability of the results will be reached via using complex approach (functional, molecular, morphological). Moreover, on the level of acute experiments, we will confront selectedbiochemical markers of perivascular adipose tissue (plasma/serum) as well as vasoactive responses of arteries isolated from normotensive and hypertensive rats with biochemical markers and reactivity of vessels isolated after nephrectomy from normotensive patients and patients with essential hypertension.

 

Study of biological effects of H2S/NO products and molecular mechanism of their actions

Principal investigator: RNDr. Karol Ondriaš, DrSc.

Principal investigator for INPP: RNDr. Soňa Čačányiová, PhD.

Lasting of the project: 1.7. 2016 – 30.6. 2020

Registration number of the project: APVV-15-0371

Anotation:

Now it is well acknowledged that endogenously produced H2S affects and is involved in regulation of many physiological and pathological functions of living organisms. It is suggested that biological effects of H2S might not result from actions of H2S alone, but from its oxidation products, which come from e.g. interaction of H2S with NO. Last four years, our “international” group indentified the following products of H2S and NO interaction: nitrosopersulfide (SSNO−), polysulfides (HSn−) and dinitrososulfite [N-nitrosohydroxylamine-N-sulfonate (SULFI/NO) (Proc Natl Acad Sci U S A. 112, 2015, E4651-E4660). Biological effects and molecular mechanisms of these products are not completely understood Therefore the aim of our project is to explore biological effects of the products of H2S/NO interactions and to study their molecular mechanism of their actions. Particularly, as a continuation of our research, we will study their effects on rat blood pressure and aortic rings relaxation. To elucidate molecular mechanisms of their biological effects, we will study their influence on expression of enzymes that endogenously produce H2S (CBS, CSE and 3-MST), on intracellular membrane channels, concentration of intracellular calcium, lipid peroxidation and their antioxidant properties. Goal of the project is also to find out, if studied compounds could provide us with information leading to a drug design based on their molecular structure, what could be an object for next application studies and lead to implementation in medical praxis.

 

VEGA

Nitroso-sulphide signal pathway – novel regulator vasoactive effects in different types of arterial hypertension

Principal investigator: RNDr. Soňa Čačányiová, PhD.

Lasting of the project: 1.1. 2018 – 31.12. 2021

Registration onumber of the project: 2/0103/18

Anotation:

Nitric oxide (NO) and hydrogen sulphide (H2S) belong to important gaseous molecules engaged to the regulation of arterial tone in normotensive conditions. NO and H2S interaction includes a formation of new products which are part of an original nitroso-sulphide signalling pathway. Our previous experiments in Wistar rats demonstrated that these new signal molecules triggered a specific vasoactive response, different from the effect evoked by NO and H2S. In condition of arterial hypertension, the vasoactive effects of the novel signalisation have not been described yet. The aim of this project is to characterise the role of NO and H2S as well as of the nitroso-sulphide signalling pathway in different animal models of hypertension: essential (SHR), NO-deficient and also metabolic syndrome (hypertriglyceridemia – HTG). A simultaneous investigation of human vessels isolated from patients with hypertension and dyslipidemia represents an appropriate way how to associate results of basic research with clinical practice.

 

Relationship between body adiposity and functional properties of arteries in rat

Principal investigator: Mgr. Anna Zemančíková, PhD.

Lasting of the project: 1.1. 2018 – 31.12. 2020

Registration number of the project: 2/0147/18

Anotation:

High levels of body mass index are considered as an important risk factor contributing to cardiovascular impairment. However, recent studies have brought some noteworthy findings that moderately increased amount of body fat is beneficial with respect to prognosis of patients with heart and vessel diseases. The aim of this project is to analyse the relationship between the level of body adiposity/weight and cardiovascular function in terms of blood pressure regulation and functional properties of selected arteries in rat. An effort will be made to reveal how the vessels are modulated by the adjacent perivascular adipose tissue (PVAT) in healthy normotensive and in hypertensive rats after moderate and considerable increase in body adiposity induced by dietetic interventions in different ontogenetic stages. Several biometric and hemodynamic parameters will be measured, as well as some biochemical parameters in blood and tissues and in vitro reactivity of arteries in the presence of PVAT and after its removal.

 

Study of critical endogenous biomarkers and signaling pathways in hypertension and cardiovascular diseases

Principal investigator: RNDr. Ima Dovinová, PhD.

Lasting of the project: 1.1. 2017 – 31.12. 2019

Registration number of the project: 2/0148/17

Anotation:

Several endogenous factors and signaling pathways contribute to the development of cardiovascular diseases. Activation of the renin-angiotensin and aldosterone system (RAAS), deteriorated relaxation of vasoactive systems producing NO and H2S, as well as an imbalance

of redox pathways producing reactive oxygen and nitrogen species (ROS a RNS). Hypertension is a major risk factors of cardiovascular diseases and leads to functional and structural alterations in blood vessel walls. The landmarks of developed hypertension are increased vasoconstriction regulated by RAAS, oxidative stress manifested in increased ROS and RNS, endothelial dysfunction (increased levels of assymetric dimethyl-arginine – ADMA – and homocystein) as well as cardiovascular remodeling (activation of metaloproteinases). Detection, monitoring, and regulation of the critical pathology markers can lead to a better management of hypertension- and metabolic syndrome-related diseases.

 

Fund of Štefan Schwarz

Project: Study of endogenous regulators of NOS and redox regulation in hypertension

Principal investigator: Mgr. Miroslava Majzúnová, PhD.

Lasting of the project: 1.5.2016 – 31.4. 2020

Anotation:

The aim of the project is the evaluation of the relationship between endogenous inhibitor of NOS (pathway ADMA-DDAH), radical pathway and antioxidant response as well as PPARgamma action in these mechanisms at the level of central and peripheral blood pressure regulation.

 

SAIA – National Scholarship Programme

Project: The role of NO-H2S interaction in arterial hypertension

Principal investigator: Muobarak Tuorkey, PhD.; Damanhour University, Egypt

Lasting of the project: 1.2. 2018 – 30.11. 2018

Anotation:

The aim of the study stay is to evaluate the role of mutual interaction among NO signalization, H2S regulatory pathway and RAS in cardiovascular system in normotensive conditions and during arterial hypertension. We will confront the vasoactive responses of arteries isolated from normotensive and spontaneously hypertensive rats (SHR), integrated response of the whole cardiovascular system (blood pressure) and selected biochemical markers (plasma, serum, perivascular adipose tissue, total vascular tissue etc.) with biochemical markers and reactivity of vessels isolated after nephrectomy from normotensive patients and patients with arterial hypertension.

 


Past projects

Study of regulation of radical and cellular signaling during hypertension and influence of novel therapies on this signaling

Project no.: APVV-0348-12

Project leader: Miroslav Barančík (Institute for Heart Research SAS)

Project leader in INPP SAS: Ima Dovinová

Duration: 1.10.2013 – 30.9.2017

Description:

Oxidative overload is an important factor in hypertension. The radicals can have negative effects on cardiovascular system associated with remodeling of the extracellular matrix, changes in cell proliferation and induction of apoptosis, while on the other hand, reactive oxygen species and nitric oxide, as intracellular signaling molecules, are also involved in activation of protein kinase signaling pathways connected with adaptive processes. An important player in processes associated with the effect of free radicals and hypertension development is the activation of intracellular signaling (kinase pathways), ABC transporters, proteins of intercellular communication and link between transcription factors and activation of antioxidant and detoxificant response through NrF2-ARE pathway. The goal of the project is to characterize the effect of short and long-term therapies on activation of the antioxidant response and overall damage of vascular and heart tissues (changes in metalloproteinases), clarification of intracellular (kinases cascades) and intercellular (connexins) signaling and enzyme regulation connected with radical formation (NOX, SOD and NOS).

 

Study of molecular mechanisms of H2S biological effects

Project no.: APVV-0074-11

Project leader: Karol Ondriaš (Institute of Molecular Physiology and Genetics SAS)

Project leader at INPP SAS: Soňa Čačányiová

Duration: 1.7.2012 – 31.12.2015

Description:

The results of the project showed that NO and H2S interacted to form new products. The chemical nature of the H2S-NO interaction is a mixture of NO, nitrsosopersulphides (SSNO) and unknown compound releasing nitroxyl (HNO). This mixture revealed different vasoactive effects than NO and H2S alone. We also showed that temporal blood pressure decrease after intravenous H2S administration could be evoked not only by activation of KATP channels but also by an inhibitory effect of H2S on renin-angiotensin system (RAS). The study of H2S effects in the condition of hypertension confirmed that spontaneously hypertensive rats have some endothelium-dependent ,,compensating” mechanisms coupled by NO a H2S signalization pathways, which are started in prehypertensive stage of essential hypertension. We also followed the effect of AP39, a new H2S donor whose mechanisms of action has not been investigated yet. We showed that in the condition of decreased bioavailability endogenous NO and H2S AP39 could represent a new therapeutic tool.

 

NO and H2S signal pathways and their interaction in the control of vascular tone during early developmental stage of experimental hypertension

Project no.: VEGA 2/0074/14

Project leader: Soňa Čačányiová

Duration: 1.1.2014 – 31.12.2017

Description:

Nitric oxide (NO) and hydrogen sulphide (H2S) belong to important gaseous molecules engaged in vascular smooth muscle regulation during normotensive as well as hypertensive conditions. The aim of the project is to characterize and compare the role of NO and H2S, their signal pathways and interaction, in young normotensive rats and in rats with developing experimental hypertension (NO-deficient and essential hypertension). Simultaneous exploration of both signal pathways on the same vessels isolated from normotensive and hypertensive rats and humans represents the actual way how to associate the results of basic research with clinical practice.

 

The role of perivascular adipose tissue in the regulation of vascular tone in rats with cardiovascular dysfunction

Project no.: VEGA 2/0202/15

Project leader: Anna Zemančíková

Duration: 1.1.2015 – 31.12.2017

Description:

Perivascular adipose tissue (PVAT) which surrounds most of the systemic arteries is considered to be an important modulator of their functions. Relaxant factors produced by this tissue inhibit vascular contractile responses to many vasoconstrictor substances. The aim of this project is to analyse the mechanisms which are responsible for this modulatory effect. In addition to the direct action of the PVAT-released substances on vascular smooth muscle we intend to study also the possible interaction of PVAT with vascular sympathetic nervous system. These interactions are important for maintaining vascular tone in physiological as well as in pathological conditions associated with vascular dysfunction, e.g. in hypertension. Our goal is to discover whether the changes in paracrine functions of PVAT may participate in the origin of vascular disorders and whether they precede the abnormalities of vascular reactivity at the level of smooth muscle.

 

The effect of nitric oxide and hydrogen sulfide on structure and function of cardiovascular system in normotenzive and hypertenzive rats

Project no.: VEGA 2/0067/13

Project leader: František Kristek

Duration: 1.1.2013 – 31.12.2016

Description:

In blood pressure regulation are, besides classical transmitters, engaged gaseous transmitters – mainly nitric oxide (NO) and new very perspective substance hydrogen sulfide (H2S). Completely new signal pathway seems to be neuronal NO synthase (nNOS) and its interactions with other regulatory systems including H2S. nNOS inhibits renin synthesis in macula densa cells and via it influences renin-angiotensin system, which is intimately related to sympathetic nervous system. Enzymes responsible for H2S production are also present in kidney and H2S is, beside others, engaged in renal blood flow and vessel wall tone regulation. The aim of the present project is to study the effect of nNOS and eNOS alone and/or in combination with H2S and other systems engaged in cardiovascular regulation during normotensive and hypertensive conditions. The availability of the results will be reached via complex functional (in vivo and in vitro), morphological, and biochemical approach.

 

Effect of PPAR gamma agonists on antioxidant response and on regulation of radical and cell signaling in hypertension

Project no.: VEGA 2/0129/14

Project leader: Ima Dovinová

Duration: 1.1.2014 – 31.12.2016

Description:

An important factor in human and experimental hypertension is the excessive oxidative load. The oxidative stress up-regulates antioxidant response (phase II response) by activation of transcription factor Nrf2. Application of substances modulating the expression of such antioxidant response may be beneficial in reduction of blood pressure and in regression against blood vessels and other tissues damage. An important role in processes associated with free radicals and development of hypertension is activation of intracellular signaling, which includes kinase pathways. The PPAR gamma agonists, used predominantly in diabetes mellitus therapy, seem to be helpful also in regulation of hypertension and are important in studying „cross-talk“ among Nrf2 a PPAR gamma. The aim of the project is to explore the effect and molecular mechanisms of short and long therm therapy of PPAR gamma agonists on antioxidant response, blood vessels and heart damage.

 

Signal pathway of nitric oxide and hydrogen sulfide, its disturbances and participation in development of hypertension and atherosclerosis

Funding: Ministry of Health of the Slovak Republic

Project number: 2012/51-SAV-1

Project leader: František Kristek

Duration: 19.7.2013-30.6.2016

Description:

Long-term blood pressure increase evokes pathological alterations in cardiovascular system and it could be the main reason for initiation of atherosclerosis. The exact cause of hypertension as well as atherosclerosis is not known. Both represent the complex of biological processes in which cooperate many regulatory systems. Besides classical transmitters there are also involved important gaseous molecules such as nitric oxide (NO) and hydrogen sulfide (H2S). They are actively engaged in regulation (reactivity of smooth muscle, structure of the arterial wall, heart and vessel trophicity, synthesis and composition of extracellular matrix) as well as in modulation of other systems. The goal of the project is to prove that there exists specific interaction between function or dysfunction of endothelium (endothelial NO synthase) and neuronal NO synthase with other systems: sympathetic nerve system, renin-angiotensin-aldosterone system (RAAS) and H2S with impact on function and ultrastructure of vessel wall (mainly ECM). Disruption of cooperation among systems could be one of the main causes of pathological responses of cardiovascular system leading to hypertension and atherosclerosis.

 


Collaboration

International

National Institute for Public Health and the Environment (RIVM) (Dr. Eugene Jansen), Utrecht, Netherlands
Characteristic: Scientific-experimental cooperation in the area of the staining of biomarkers and markers of oxidative stress.

 

Clinical Institute of Medical and Chemical Laboratory Diagnostics (Dr. Andreas Meinitzer), Gratz, Austria
Characteristic: Scientific-experimental cooperation in the area of the detection of endogenous NO-synthase inhibitor (ADMA, SDMA).

 

University of Catania, Department of Biomedical and Biotechnological Sciences (Dr. V. Calabrese), Catania, Italy
Characteristic: Scientific-experimental cooperation from 2013. Realization of study stay in the frame of National Scholarship Programme aimed on the evaluation of PRAR gamma agonists´s effects on antioxidant response of Nrf2/ARE pathways in neurodegenerative models and Diabetes mellitus (Dr. Majzúnová, 2013-2014).

 

Universitätsmedizin der Johannes Gutenberg-Universität Mainz (Dr. A. Daiber), Mainz Germany
Characteristic: Scientific-experimental cooperation from 2013. Realization of study stay in the frame of National Scholarship Programme aimed on the evaluation of different markers of redox balance in different pathological stages (Ing. Kvandová, 2017-2018).

 

National

Urological clinic of Acad. L. Derer Hospital, Medical faculty of Comenius University, Bratislava( Dr. J. Breza)
Characteristic: The cooperation allows the research of selected physiological mechanisms and biochemical parameters on samples of human material, vessels isolated after nephrectomy from normotensive and hypertensive patients.

 

Long-term cooperation with Institute of Clinical and Translational Research BRC SAS, Faculty of Natural Sciences, Pharmaceutical faculty, Faculty of Medicine, Comenius University, Faculty of Chemical and Food Technology, Slovak University of Technology, Bratislava in the frame of research projects and teaching.