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

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 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.

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.