Effects of uwhangchungsimwon on cell viability, proliferation, and gene expression of human neuronal cell line IMR32

Uwhangchungsimwon (pill, UC) is one of the traditional Korean medical prescriptions that has been most frequently used for stroke. To characterize the effects of UC on human neuronal cells, the human neuroblastoma cell line IMR32 was treated with UC, and cell viability, cell proliferation, apoptosis, and gene expression were analyzed. The effect of UC on recovery of cell viability was analyzed following stress induction by nutrient depletion or cold shock. Flow cytometric analysis of the cell cycle showed that UC inhibits cell cycle progression of IMR32 in a dose- and time-dependent manner. UC was also identified to increase cell viability and suppress apoptosis induction by a DNAdamaging agent, etoposide. Quantitative RT-PCR analysis revealed that expressions of thep53 tumor suppressor gene and its downstream effect, Wafl, are stimulated whereas expressions of positive cell cycle regulators, c-Myc, c-Fos, and Cyclin DJ were repressed by UC treatment. Moreover, while expression levels of apoptosis inhibitors, Bcl-2 ana Bel-XL were increased following UC treatment, that of an apoptosis promoter, Box, was decreased. In addition, expression of BMP-7, which has been recently demonstrated to improve the motor neuron recovery from stroke, was induced by UC while it was not detected in untreated cells. Taken together, our data suggest that the pharmacoclinical effects of UC might be derived in part from its negative regulation of cell proliferation and apoptosis through the transcriptional control of related genes.