ABSTRACT
Human mesenchymal stem cells (MSCs) may be used in cell-based therapy to promote neovascularization for the treatment of ischemic diseases. However, high levels of reactive oxygen species (ROS) derived from the pathophysiological ischemic environment induce senescence and apoptosis of MSCs, resulting in reduced functionality and defective neovascularization. Therefore, the present study aimed to determine the protective effects of Cirsium setidens, a natural product, on oxidative stressinduced apoptosis in MSCs. The present study investigated for the change of ROS levels in MSCs using ROS assays. In addition, cell viability determined by MTT and TUNEL assays. Western blot analysis was performed to investigate the change of apoptosisassociated proteins in MSCs. Treatment of MSCs with hydrogen peroxide (H2O2; 200 µM) significantly increased intracellular ROS levels and cell death; however, pretreatment with C. setidens (100 µg/ml) suppressed H2O2induced ROS generation and increased the survival of MSCs. H2O2induced ROS production increased the levels of phosphorylatedp38 mitogen activated protein kinase, cJun Nterminal kinase, ataxia telangiectasia mutated and p53; these increases were inhibited by pretreatment with C. setidens. In addition, C. setidens inhibited ROSinduced apoptosis of MSCs by increasing the expression levels of the antiapoptotic protein Bcell lymphoma 2 (BCL2), and decreasing the expression levels of the proapoptotic protein BCL2associated X protein. These findings indicated that pretreatment of MSCs with C. setidens may prevent ROSinduced oxidative injury by regulating the oxidative stressassociated signaling pathway, and suppressing the apoptosisassociated signal pathway. Therefore, C. setidens may be developed as a beneficial broadspectrum agent for enhancing the effectiveness of MSC transplantation in the treatment of ischemic diseases.