Identification of cytotoxic markers in methamphetamine treated rat C6 astroglia-like cells

Abstract

Methamphetamine (METH) is a powerfully addictive psychostimulant that has a pronounced effect on the central nervous system (CNS). The present study aimed to assess METH toxicity in differentiated C6 astroglia-like cells through biochemical and toxicity markers with acute (1 h) and chronic (48 h) treatments. In the absence of external stimulants, cellular differentiation of neuronal morphology was achieved through reduced serum (2.5%) in the medium. The cells displayed branched neurite-like processes with extensive intercellular connections. Results indicated that acute METH treatment neither altered the cell morphology nor killed the cells, which echoed with lack of consequence on reactive oxygen species (ROS), nitric oxide (NO) or inhibition of any cell cycle phases except induction of cytoplasmic vacuoles. On the other hand, chronic treatment at 1 mM or above destroyed the neurite-like processors and decreased the cell viability that paralleled with increased levels of ROS, lipid peroxidation and lactate, depletion in glutathione (GSH) level and inhibition at G0/G1 phase of cell cycle, leading to apoptosis. Pre-treatment of cells with N-acetyl cysteine (NAC, 2.5 mM for 1 h) followed by METH co-treatment for 48 h rescued the cells completely from toxicity by decreasing ROS through increased GSH. Our results provide evidence that increased ROS and GSH depletion underlie the cytotoxic effects of METH in the cells. Since loss in neurite connections and intracellular changes can lead to psychiatric illnesses in drug users, the evidence that we show in our study suggests that these are also contributing factors for psychiatric-illnesses in METH addicts.

Toxic Potentials of Methamphetamine in Neuroblastoma Cell Line and its Effects on the Mitochondrial Activity

“…ROS productions are potent initiator of cell death. as suggested by Badisa et al [18], lack of cell death following acute METH treatment infer lack of ROS production and nitric oxide generation or cell cycle inhibition [18]. Only the viability of cells treated with 10 µg/mL after 48 hours, was below the benchmark, suggesting that METH has a neurotoxic effect that affects viability since 93% suggests that cells are not well viable.…”

Central Nervous System Stimulants Limit Caffeine Transport at the Blood–Cerebrospinal Fluid Barrier

“…In human autopsy cases, the stimulants are used as neurostimulants; however, in culture experiments, 4-AP is used instead of stimulants. Studies have shown that some cells do not respond to the administration of stimulants into cultured cells unless a large amount of the stimulant is used in the cultured cells, and thus, 4-AP was used [20][21][22]. Although 4-AP is a potassium blocker [23,24], as mentioned earlier, caffeine is a highly lipophilic substance [13] and can easily pass through cell membranes, so we thought that the BCSFB would not be susceptible to the effects of 4-AP.…”

The Role of HSP90α in Methamphetamine/Hyperthermia-Induced Necroptosis in Rat Striatal Neurons

“…In humans, the common dose of a well-adopted abuser is 1 g or more METH per day ( Simon et al, 2002 ; Melega et al, 2007 ). In these cases, the blood concentration of METH may increase to the millimolar range ( Badisa et al, 2019 ). Meanwhile, systemic responses, such as immune responses and HT, might play crucial roles in METH-induced toxicity in vivo ( Papageorgiou et al, 2019 ; Marco et al, 2021 ).…”