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

Ramesh B Badisa,Kesa Randell,Carl B Goodman,Maryam Agharahimi,Selina F Darling-Reed,Chantel Wiley,Karam F A Soliman,Lekan M Latinwo

doi:10.1038/s41598-019-45845-1

Ramesh B Badisa, Kesa Randell + Show 6 more

Open Access

https://doi.org/10.1038/s41598-019-45845-1

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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.

Highlights

METH, an amphetamine derivative, is one of the popular synthetic illegal psycho-stimulants abused in the US1 and worldwide[2,3]
The cytotoxic markers we focused on include vacuolation, viability, reactive oxygen species (ROS), nitric oxide (NO) release, morphology, lipid peroxidation, lactate release, GSH level and apoptosis at acute and chronic treatments
METH did cause an induction of cytoplasmic vacuoles (n = 12, F = 4.5, P < 0.05), which is an indication of cell injury, in the treated cells compared to the control (Fig. 1); METH did not trigger significant www.nature.com/scientificreports alteration in any cellular parameters, such as cell viability (Fig. 2A) or ROS production (Fig. 2B) or NO release (Fig. 2C) or cell cycle progression (Fig. 2D)

Summary

Introduction

METH, an amphetamine derivative, is one of the popular synthetic illegal psycho-stimulants abused in the US1 and worldwide[2,3]. Ice and speed are some popular club names for METH, and it is portrayed as a poor man’s cocaine and the drug of choice for economically poor addicts Though both METH and cocaine work as CNS stimulants, METH triggers much stronger pharmacological effect[9,10] than cocaine because the latter is metabolized most rapidly and disposed of the body in contrast to the former[9]. The primary aim of our study is to discern the cytotoxic markers for METH using rat C6 astroglia-like cells. We tested these cells at acute (1 h) and chronic (48 h) time points. The inhibitory role of METH on cell cycle phases was assessed

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