Cocaine induces HIF1 (Hypoxia Inducible Factor1) in rat C6 glial cells (LB619)

Abstract

Investigations with astroglial cells carry more prominence in drug abuse studies. However, not many investigations have been conducted on the toxicity of various abused drugs, like cocaine. The present study was aimed to discern the effect of cocaine on rat astroglioma cells. Glial cells were grown in 10% FBS (Fetal Bovine Serum) in Dulbecco’s modified Eagle medium (DMEM) and penicillin/streptomycin (100 Units/0.1 mg per ml). Treatment of glial cells with cocaine for 24 and 48 hours caused significant loss of cell viability. We also used rat cytokine array membranes to determine the change in rat C6 glial cell cytokine expression upon cocaine exposure (2, 4 and 7 mM). We found after 24 hours cocaine exposure, expression of vascular endothelial growth factor (VEGF) was relatively high in each concentration compared to control and these results were confirmed with western blot analysis. Preliminary data from different studies indicated that hypoxia is a potent inducer of VEGF in vitro partly because of an increased transcription rate, mediated by binding of hypoxia‐inducible factor‐1 (HIF1) to a hypoxia responsive element, VEGF gene. Thus following cocaine treatment of various concentration and RNA isolation, real time PCR (Polymerase Chain Reaction ) was carried out using the IQ Sybr Green Supermix in conjunction with MyiQ software (Bio‐Rad, Hercules, CA). Expression of HIF1 followed by VEGF observed in each concentration compared to control. We concluded that cocaine may be involved in hypoxia in C6 glial cells.Grant Funding Source: This research was supported by National Institute on Minority Health Disparities of the National Institute of Health under award number G12MD007582, FAMU RCMI, Pharmaceutical Research Center