Naslov
Detection of early apoptosis in neonatal rat cardyomyocyte culture

Autori
Kniewald, Hrvoje ; Polančec, Denis ; Malčić, Ivan ; Radošević, Kristina ; Gaurina-Srček, Višnja ; Kniewald, Jasna ; Kniewald, Zlatko

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstracts Book of The Fourth World Congress of Pediatric Cardiology and Cardiac Surgery, September 18-22, 2005, Buenos Aires, Argentina / Capelli, H. ; Kreutzer, G. - Buenos Aires : The Argentine Community of Pediatric Cardiologists and Cardiovascular Surgeons, 2005

Skup
THe Fourth World Congress of Pediatric Cardiology and Cardiac Surgery

Mjesto i datum
Buenos Aires, Argentina, 18.09.2005. - 22.09.2005

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
cardiomyociytes; apoptsis; neonatal rat

Sažetak
Myocardial sympathetic activity is increased in heart failure, and norepinephrine (NE)- stimulate apoptosis of cardiomyocytes through α - and β -adrenergic pathway. Isoprotenerol (ISO), as well as NE, induce apoptosis in neonatal rat cardiomyocyte (NRC) culture. Apoptosis has been demonstrated to occur in the myocardium in a variety of pathological situations. The number of apoptotic myocytes is increased in myocardium obtained from patients with end-stage heart failure and myocardial infarction and in myocardium from experimental models of myocardial hypertrophy and failure, including the aortic-banded rat, the spontaneously hypertensive rat, rats with myocardial infarction, and dogs with pacing-induced failure. Experiments with cultured cardiac myocytes have demonstrated that apoptosis can be stimulated in vitro by several endogenous peptides that are increased in the hypertrophied or failing myocardium, including TNF-α , angiotensin II and atrial natriuretic peptide, as well as with ISO and NE. Primary ventricular cardiac myocytes were prepared from 6 ­8 days old Fisher rats. Cells were plated to 24-well culture plates at density of 1, 25x105 cells/cm2 in DMEM with 10% FBS. NRC were maintained at 37oC in humidifier air with 5% CO2. After 48 hours cells were treated with NE (1 and 10 mcmol/L), E (1 and 10 mcmol/L) and ISO (1 and 10 mcmol/L). 24 hours after treated cells were harvested and counted. Detection of the early stage s of apoptosis was performed using JC ­1, an mitochondrial depolarization sensitive dye, and flow cytometry. NRC treated with NE (10 mcmol/L), E (10 mcmol/L) and ISO (10 mcmol/L) had significantly higher apoptotic rate detected by JC ­1 compared to control group. Lower concentrations of NE, E and ISO did not induce increase in constitutive apoptosis compared with control group, i.e. detected changes in Δ Ψ m were at the same level as in control. JC1 staining and flow cytometric analysis were succcesfuly used for detection of early apoptotic event s in primary NRC. Disrupted Δ Ψ m was significantly augmented after induction of NRCs apoptosis through the adrenergic pathway . It should be further determinated whether this disruption of Δ Ψ m is an early requirement for apoptosis or a consequence of the apoptotic ­signaling pathway. This work has been supported by the Ministry of Science, Education and Sport Republic of Croatia (Grants no. 0058001 and 0058010) and Pliva inc.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti, Biotehnologija

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