Reactive oxygen species (ROS) mediate vascular cell dysfunction and lead to atherosclerosis and other chronic cardiovascular diseases. The root of Pueraria lobata(Willd.) Ohwi, also known as kudzu or Gegen (Chinese name), is one of the most important herbs in traditional Chinese medicine and has been widely used in the treatment of cardiovascular diseases, diabetes, osteonecrosis and neurodegradation diseases. In this study, an ethanol extract from kudzu root was prepared and the in vitro protective effect of the kudzu root extract (KUD) on human umbilical vein endothelial cells (HUVECs) was investigated.
An ethanol extract of dried kudzu root was purified with an AB-8 resin column, and the concentrations of puerarin, daidzin and daidzein in the KUD were determined using UV spectroscopy. HUVECs were pretreated with various concentrations of the KUD with or without rotenone and the viability was assessed by AlamarBlue cell viability assay. Next, HUVECs were pretreated with the KUD and then treated with rotenone, and the levels of ROS generation, apoptosis, and changes of the mitochondrial membrane potential (ΔΨm) in HUVECs were measured using fluorescent staining assay and high-content analysis.
... 2.1. Reagents. Ferric chloride (FeCl 3 ), DMSO, and N-acetyl-cysteine (NAC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Puerarin, daidzin and daidzein standards at purities greater than 98% were purchased from Biopurify Phytochemicals Ltd. (Chengdu, China). ...
