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ORAL PRESENTATION / TAM METİN SÖZLÜ SUNUM

Therefore, we hypothesized that European Blueberry (Vaccinium myrtillus L.) seed oil
(BSO) exerts cardioprotective effects via potential anti-inflammatory and anti-apoptotic

mechanisms in an H9c2 cell model of doxorubicin-induced cardiotoxicity.

MATERIAL AND METHODS

Essential Oils: European Blueberry (Vaccinium myrtillus L.) seed oil and its
compositional analysis were supplied by Art de Huile (Istanbul, Turkey).

H9c2 Cell Culture: The H9c2 (Rat, Heart, myoblast) cell line was obtained from the
American Type Culture Collection (ATCC, USA). The cryotubes containing the cell lines stored

in liquid nitrogen were taken out of the tank and briefly thawed in a 37°C water bath. The
cells were then transferred to T75 cm flasks. After 48 h, H9c2 cells were counted at a
2
5
concentration of 2 × 10 cells per well in DMEM with 10% FBS, seeded into a 96-well plate,
and maintained at 37°C in a humidified environment with 5% CO2. After 24 h, the cells were
exposed to varying concentrations of BSO (1–640 µg/ml) dissolved in 0.1% dimethyl

sulfoxide (DMSO).
Cell viability assay: The anti-proliferative effect of BSO on H9c2 cells was evaluated

using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay after
5
culturing at a density of 2 × 10 cells/well in 96-well plates for 24 h. The absorbance values
at 570 nm were measured three times using a microplate reader spectrophotometer (Epoch
Microplate Spectrophotometer, BioTek, USA), following the manufacturer’s instructions

(Roche, Germany). A total of 10 µl (5 mg/ml) of MTT solution was added to each well, and
which the plate was incubated at 37 °C for 4 h. To each well of the plate, 100 µL of

solubilization buffer (DMSO) was carefully added and the plate was then incubated at 37°C

for 10 min. This formula was used to determine the suppression of cell proliferation.
( ) − ( )
% Growth inhibition = 100 − × 100
OD (control) − OD (blank)

Treatment Groups: The experimental groups were designed as follows.
▪ Control group: no treatment.

▪ DOX group: H9c2 cells exposed to 5 µM doxorubicin for 24 h
▪ BSO 10 + DOX: Cells were pretreated with 10 µg/ml BSO for 30 min and then treated

with 5 µM doxorubicin for 24 h.

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