Učinki fluvastatina in valsartana na sistem renin-angiotenzin v primarnih človeških endotelijskih celicah koronark, gojenih in vitro

Kovačević, Sanja (author), Černe, Darko (mentor)

Pri razvoju ateroskleroze in srčno-žilnih bolezni ima pomembno vlogo sistem renin-angiotenzin (RAS). Angiotenzin II (Ang II), glavni mediator tega sistema, je visoko izražen v žilni steni. Ang II v arterijski steni sodeluje pri okvari endotelijske funkcije in razvoju aterosklerotičnega plaka. Za preventivo in zdravljenje srčno-žilnih bolezni se uporabljajo statini in inhibitorji RAS, ki imajo poleg primarnih učinkov na znižanje holesterola in krvnega tlaka, tudi protivnetne in antioksidativne pleiotropne učinke, ki vodijo v izboljšanje žilne funkcije. Namen naloge je bil razviti celični model za proučevanje subterapevtskih učinkov fluvastatina in valsartana na celični RAS v primarnih človeških endotelijskih celicah koronarnih arterij (HCAEC). Celice smo po aplikaciji zdravilnih učinkovin tretirali s serumskim amiloidom A (SAA) in določili vpliv SAA in zdravilnih učinkovin na izražanje izbranih genov sistema RAS. Ugotovili smo, da v nasprotju s pričakovanji in literaturnimi viri v HCAEC celični RAS ne obstaja, saj ni izražanja prekurzorja angiotenzinogena (AGT) ter encimov, ki sodelujejo pri njegovi pretvorbi v aktivne mediatorje RAS. Rezultati so pokazali, da SAA ne vpliva na izražanje komponent RAS. Izjema je encim angiotenzin konvertaza (ACE), katerega izražanje SAA celo zniža. V HCAEC se izraža gen za angiotenzinski II receptor tipa 1 (AGTR1), kar bi lahko bilo pomembno, če se Ang II iz krvi po receptorski endocitozi ne razgradi popolnoma in sproščen intracelularno deluje na znotrajcelične receptorje AGTR1 ter tako povzroča učinke RAS intracelularno. SAA deluje proaterogeno, saj poveča izražanje gena za žilno-celično adhezijsko molekulo 1 (VCAM-1), provnetni citokin interlevkin 6 (IL-6), kemokin interlevkin 8 (IL-8) in encim matriks metaloproteinazo 1 (MMP1). Nasprotno pa fluvastatin deluje zaščitno na endotelijske celice, saj poveča izražanje gena za encim endotelijsko sintazo dušikovega oksida (NOS3) ter zmanjša izražanje VCAM-1, IL-6 in IL-8. Ugotovili smo, da ima valsartan manjši vpliv na izražanje izbranih genov kot fluvastatin. Ker so rezultati za fluvastatin ter kombinacijo učinkovin podobni in so učinki valsartana na izražanje izbranih genov majhni, lahko sklepamo, da so ti učinki posledica delovanja fluvastatina. Glede na dobljene rezultate kombinacija učinkovin ne izkazuje sinergističnega delovanja. Našteti učinki SAA, fluvastatina in valsartana na HCAEC najverjetneje niso posredovani preko celičnega RAS, temveč so posledica aktivacije drugih signalnih poti.

• Language: Slovenian
• Keywords: ateroskleroza, celični RAS, HCAEC, SAA, fluvastatin, valsartan
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2019
• PID: 20.500.12556/RUL-108338
• Publication date in RUL: 28.06.2019

Secondary language

• Language: English
• Title: Effects of fluvastatin and valsartan on the renin-angiotensin system within in vitro cultured human primary coronary artery endothelial cells

Abstract

The renin-angiotensin system (RAS) plays an important role in the development of atherosclerosis and cardiovascular diseases. Angiotensin II (AngII), the main mediator of this system, is highly expressed in the vascular wall. In the artery wall, Ang II promotes endothelial dysfunction and the development of an atherosclerotic plaque. For the prevention and treatment of cardiovascular diseases, statins and RAS inhibitors are used, which besides the primary effect on cholesterol and blood pressure reduction, also have antiinflammatory and antioxidant pleiotropic effects that lead to improvement in vascular function. The aim of this study was to develop a cell model for the study of subtherapeutic effects of fluvastatin and valsartan on cellular RAS in primary human coronary artery endothelial cells (HCAEC). After the administration of active pharmaceutical ingredients, the cells were treated with serum amyloid A (SAA) and the effect of SAA and active pharmaceutical ingredients on the expression of selected genes of the RAS were determined. We found that, contratry to expectations and literary sources, cellular RAS does not exist in HCAEC since there is no expression of precursor angiotensinogen (AGT) and enzymes that are involved in its conversion into active mediators of RAS. The results showed that SAA does not affect the expression of RAS components. The exception is the angiotensin converting enzyme (ACE), expression of which was even reduced by SAA. HCAEC express angiotenisn II type 1 receptor (AGTR1), which may be important if Ang II is not fully degraded after the receptor endocytosis and released intrucellularly where it binds to intracellular AGTR1 receptors. SAA acts proaterogenic as it increases the expression of the vascular cell adhesion molecule 1 (VCAM-1), cytokine interleukin 6 (IL-6), chemokine interleukin 8 (IL-8) and matrix metalloproteinase 1 (MMP1). On the other hand, fluvastatin has protective effects on endothelial cells, as it increases the expression of the enzyme endothelial nitric oxide synthase (NOS3) and reduces the expression of VCAM-1, IL-6 and IL-8. Furthermore our results also show that valsartan has a lower impact on the expression of selected genes than fuvastatin. Since the results for fluvastatin and combination of drugs are similar and the effects of valsartan on the expression of selected genes are small, we can conclude that these effects are result of the action of fluvastatin. Based on the results obtained, the combination of active substances does not show synergistic effects. The listed effects of SAA, fluvastatin and valsartan on HCAEC are most likely not mediated by cellular RAS, but are the result of the activation of other signaling pathways.

• Keywords: atherosclerosis, cellular RAS, HCAEC, SAA, fluvastatin, valsartan