PhD Scientific Days 2017

Oral Presentations: Cardiovascular

Investigation of simulated ischemia/reperfusion injury in an in vitro hyperlipidemic comorbidity model

Előadó neve

Dr. Makkos, András

Előadó munkahelye

Semmelweis University Department of Pharmacology and Pharmacotherapy

Előadó telefonszáma

+36703707324

Előadó e-mail címe

makkos.andras@med.semmelweis-univ.hu

Az előadás címe

Investigation of simulated ischemia/reperfusion injury in an in vitro hyperlipidemic comorbidity model

Szerző(k) neve és munkahelye

András Makkos1
1 PhD student, Semmelweis University Department of Pharmacology and Pharmacotherapy, Budapest

Szekció

Oral Presentations: Cardiovascular

Data of the presenter

Doctoral School: Pharmaceutical Sciences
Program: Experimental Cardiovascular Pharmacological Research
Supervisor: Prof. Peter Ferdinandy

Text of the abstract

Metabolic diseases, like hyperlipidemia, metabolic syndrome are risk factors of ischemic heart diseases and worsen its prognosis. Our research group set up an in vivo hypercholesterolemic rat model previously. Although, cell culture models are suitable for exploration of intracellular signaling pathways in hypercholesterolemia.

Our aim was to investigate the effect of simulated ischemia-reperfusion injury (sI/R) on myocardial cells under hypercholesterolemic conditions.

Cardiomyocytes were isolated from young adult wistar rats and cultured for 2 days in control or cholesterol containing medium with different concentrations (group M1, M2, M3). We applied 30 min simulated ischemia (lactate containing hypoxic solution + 95% N2 and 5% CO2 atmosphere) and 2 hours of reperfusion (M1-3 + normoxic conditions). Normoxic control groups were also used. After the sI/R we measured the cell viability with calcein staining and the total cell count with digitonin + propidium-iodide. For the measurement the level of oxidative stress, dihydroethidium (superoxid levels) and 2’,7’-dichlorodihydrofluorescein-diacetate staining (for total reactive oxygen species - ROS) were applied.

Our result are expressed in percentage compared to the control groups. The viability of the cells were decreased in a cholesterol concentration-dependent manner after sI/R (M1: 70±18%, M2: 54±14%, M3: 53±5%) and also in the normoxic groups (M2: 78±24%, M3: 83±23%). The oxidative stress measurements showed elevated superoxide levels in sI/R groups (M2: 351±117%, M3: 295±23%) Elevated ROS level was observed in normoxic (M2: 162±26%, M3: 171±62%) and one of the sI/R groups (M1: 307±195%). For statistics one-way ANOVA (p<0.05) and Tukey posthoc test was used.

The viability of the cardiomyocytes was reduced and the oxidative stress level was elevated under hypercholesterolemic conditions and found to be concentration dependent. sI/R could provoke further cell damage, which was also cholesterol concentration dependent. Based on these findings present in vitro comorbidity model can be suitable for testing drug candidates.

Kind

Szabad

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

1293

Authors (legacy)

András Makkos1
1 PhD student, Semmelweis University Department of Pharmacology and Pharmacotherapy, Budapest