ABSTRACT
BACKGROUND: Chrysophanic acid, also known as chrysophanol, has a number of biological activities. It enhances memory and learning abilities, raises superoxide dismutase activity, and has anti-cancer effects in several model systems. According to previous reports, chrysophanic acid-induced cell death shares features of necrotic cell death. However, the molecular and cellular processes underlying chrysophanic acid-induced cell death remain poorly understood. METHODS: Chrysophanic acid-induced cell death was monitored by cell viability assay and Annexin V-propidium iodide (PI) staining of renal cell carcinoma Caki-2 cells. The induction of intracellular reactive oxygen species (ROS) by chrysophanic acid and the suppression of ROS by anti-oxidants were evaluated by 2',7'-dichlorofluorescin diacetate staining. The expression and phosphorylation of proteins that are involved in apoptosis and necroptosis were detected by immunoblotting. RESULTS: The extent of chrysophanic acid-induced cell death was concentration and time dependent, and dead cells mainly appeared in the PI-positive population, which is a major feature of necrosis, upon fluorescence-activated cell sorting analysis. Chrysophanic acid-induced cell death was associated with the generation of intracellular ROS, and this effect was reversed by pretreatment with N-acetyl cysteine. Chrysophanic acid-induced cell death was not associated with changes in apoptotic or necroptotic marker proteins. CONCLUSIONS: The cell death induced by chrysophanic acid resembled neither apoptotic nor necroptotic cell death in human renal cell carcinoma Caki-2 cells.
Subject(s)
Humans , Apoptosis , Carcinoma, Renal Cell , Cell Death , Cell Survival , Cysteine , Flow Cytometry , Immunoblotting , Learning , Memory , Necrosis , Phosphorylation , Reactive Oxygen Species , Superoxide DismutaseABSTRACT
Objective To establish a method for the determi nation of emodin and chrysophanic acid in Compound Rhubarb Spray by HPLC.Methods The ODS column was applied.methanol∶water(87∶13)served as mobile phase.The detectio n wavelength was at 254nm and the flow r ate was 0.65ml /min.Results The average recovery of emodin was 99.48%with relative standard deviation being 1.75%(n=9)and the average recovery of chrysoph anic acid was 101.46%with rela-tive standard deviation being 2.85%(n=9).Conclusion The method was simple,rapid and reli able.It is suitable for the quality control of Compound Rhubarb Spray.