Cardioprotection of Atractylodes lancea against Hypoxia/ Reoxygenation-Injured H9c2 Cardiomyoblasts
Punnee Nusuetrong PhD*, Orapin Gerdprasert DVM, PhD**
Affiliation : * Department of Physiology, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand ** Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand
Background : Atractylodes lancea (Thunb) DC has been widely used as traditional medicine in many countries including
Thailand for the treatment of fever, common cold and sore throat.
Objective : To evaluate the cardioprotective effects of Atractylodes lancea extracts against hypoxia/reoxygenation (HR)-
injured H9c2 cardiomyoblasts.
Material and Method: For cytotoxic determination, the H9c2 cells were incubated with the ethanolic extract of Atractylodes
lancea (AL-E) and water extract (AL-W) at the concentrations between 0.01-1 mg/mL in normoxia for 6, 18, 24, and 48 h. Cell
viability were determined by MTT assay and observed cellular morphology under phase contrast microscopy. In a time-
course study of HR model on H9c2 cardiomyoblasts, the cells were exposed to hypoxic condition at various time points (0.5,
1, 2, 4, 6, and 8 h) before 24 h reoxygenation. According to more than 90% of cell death, 6 h exposure to hypoxia was used
for cardioprotective evaluation throughout the present study. Cell viability, DNA condensation by Hoechst 33342, and protein
expression of ERK1/2, p-ERK1/2 and HO-1 by western blot analysis were investigated.
Results : Incubation of AL-E and AL-W at concentrations up to 1 mg/mL for 24 h showed no toxicity on H9c2 cells. Exposure
of the H9c2 cells to HR showed a time-dependent decrease in cell viability. Treatment of both AL-E and AL-W (0.05 and 0.1
mg/mL) showed protection against cell death and cellular shrinkage as well as inhibited DNA condensation. AL at the same
concentrations increased the expression of ERK1/2, p-ERK1/2 and HO-1, when compared to HR-injured cells.
Conclusion : AL protected HR-damaged H9c2 cells by inhibiting cell death, cellular shrinkage and DNA condensation, which
was partially through restoring ERK1/2, p-ERK1/2, and HO-1 expression. The present results are beneficial use of AL as
alternative medicine.
Keywords : Atractylodes lancea, Cardioprotection, Hypoxia/reoxygenation, DNA condensation, ERK, HO-1
