Cardioprotection of Atractylodes lancea against Hypoxia/ Reoxygenation-Injured H9c2 Cardiomyoblasts

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.

To evaluate the cardioprotective effects of Atractylodes lancea extracts against hypoxia/reoxygenation (HR)-injured H9c2 cardiomyoblasts.

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 timecourse 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.

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.

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.