Objective
To explore the toxicity of acrylamide on motor neuron VSC4.1 cells and relevant mechanism.
Methods
Acrylamide (20, 60, 100 g/L) was added into the culture medium of VSC4.1 cells, and the cells without acrylamide exposure were treated as controls. After 48 h, cell viability, staining, and apoptosis were evaluated. C/EBP-homologous proteins (CHOP), cysteine-containing aspartate-specific protease 12 (Caspase 12) and the unfolded protein response related gene (GRP78, ATF6, IRE1) were also examined. Meanwhile, the effects of endoplasmic reticulum stress inhibitor, 4-phenylbutyric acid (4-PBA), and antioxidant, taurine, on the VSC4.1 cells intoxicated with acrylamide were observed.
Results
The cell viability was decreased with higher acrylamide concentrations, and the survival rates of the 20, 60, and 100 g/L acrylamide groups were (88.5±6.2)%, (59.3±4.1)%, and (37.6±3.4)%, respectively (χtrend2=13.04, P < 0.05), compared with the control group (P < 0.05). Nucleus pycnosis, strong blue fluorescence, and characteristic morphology of cell apoptosis were observed using Hochest 33258 staining after acrylamide exposure. The apoptosis rates of the 20, 60, and 100g/L acrylamide groups were (9.8±0.9)%, (27.4±2.3)%, and (53.1±4.8)%, respectively, compared with the control group (χtrend2=6.98, P < 0.05). The protein expression levels of CHOP and Caspase 12 were decreased markedly in each treated group compared with the control group (P < 0.05) except the Caspase 12 of the 20 g/L acrylamide group. After 20, 60, and 100 mg/L 4-PBA were added into the VSC4.1 cells intoxicated with 60 g/L acrylamide, the cell viabilities were (125±9)%, (154±12)%, and (222±20)%, respectively, of the cells without 4-PBA treatment (P < 0.05). After 20, 60, and 100mg/L taurine were added into the VSC4.1 cells intoxicated with 60 g/L acrylamide, the cell apoptosis rates were (75±8)%, (60±6)%, (35±4)%, respectively, of the cells without taurine treatment (P < 0.05). The expression levels of 3 CHOP (GRP78, ATF6, IRE1) were also decreased with higher levels of taurine (P < 0.05).
Conclusion
Acrylamide could induce apoptosis in VSC4.1 cells and the relevant mechanism may be associated with endoplasmic reticulum stress.
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