Cocaine induces alterations in mitochondrial membrane potential and dual cell cycle arrest in rat c6 astroglioma cells.
Investigations with astroglial cells carry more prominence in drug abuse studies. However, due to earlier perception that astroglial cells were only passive bystanders in neural signal transmission, not many investigations were 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 and analyzed qualitatively for morphological features as well as vacuolation by phase contrast microscope, quantitatively for cytotoxicity, mitochondrial membrane potential by rhodamine- 123 fluorometric assay, and cell cycle analysis by flow cytometry. Based on population cell doubling time studies, glial cells were grown in 10% FBS in RPMI 1640 medium and treated with cocaine for 24 or 48 h. Microscopic assessments clearly demonstrated massive vacuolation and significant disruption at general architecture of glial cell morphology with cocaine. Chronic cocaine treatment (24 or 48 h) caused significant loss of cell viability. The sublethal dose of cocaine was found to be 4.307 and 3.794 mM at 24 and 48 h, respectively. Cocaine reduced the mitochondrial membrane potential in a dose dependent manner with ED(50) of 4 mM after 24 h. Cell cycle analysis suggested dual inhibition at G0/G1 and G2/M phases after 24 and 48 h, respectively. In summary, our findings suggest that cocaine toxicity was due to loss of mitochondrial membrane potential, vacuolation, and dual inhibition of cell cycle phases. These results may shed light in understanding the onset of some early key events in cocaine-induced toxicity in glial cells.
Badisa
Ramesh B
RB
Science Research Center, College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Room # 308E, Tallahassee, FL 32307, USA.
Darling-Reed
Selina F
SF
Goodman
Carl B
CB
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001254:Astrocytoma; D002453:Cell Cycle / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003042:Cocaine / Q000494:pharmacology*; D016195:G2 Phase / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects; D051381:Rats; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D014617:Vacuoles / Q000187:drug effects
D003042:Cocaine