Objective To observe the effects of vitamin E, vitamin C, and taurine on energy metabolism damage of human neuroblastoma cells (SH-SY5Y) induced by benzoapyrene (BaP) exposure.
Methods SH-SY5Y cells were treated with DMSO, BaP (1 μmoL/L), BaP (1 μmoL/L) + vitamin E (50 μmol/L), BaP (1 μmol/L) + vitamin C (100 μmol/L), and BaP (1 μmol/L) + taurine (100 μmol/L), respectively. After 24 h of treatment, cell viability was measured by MTT assay, reactive oxygen species (ROS) level was detected with ROS detection kit, cellular malondialdehyde (MDA) content was assessed with MDA assay kit, superoxide dismutase (SOD) activity was evaluated with SOD typing test kit, and mitochondrial respiratory and glycolysis of the cells were detected with Seahorse XFp cell energy analyzer.
Results The cell viability (84.2%±1.2%) and SOD activity(60.19±1.02) U/mg were significantly lower, while the ROS level (60.73%±3.15%) and MDA content(0.837±0.104) μmol/g were significantly higher in the BaP-treated group than in the vehicle control group100%±3.74%, (67.37±0.78) U/mg, 31.27%±1.40%, and (0.382±0.083) μmol/g, respectively (P < 0.05). The basal oxygen consumption rate (OCR)(2.64±0.43) pmol/min, ATP-linked OCR (2.09±0.32) pmol/min, maximal respiration (3.42±0.03) pmol/min, and spare respiratory capacity (0.78±0.44) pmol/min in the BaP-treated group were lower than those in the vehicle control group (4.89±0.43, 4.16±0.24, 7.06±0.11, and 2.18±0.35) pmol/min, respectively (P < 0.05), while the proton leak OCR(0.55±0.16) pmol/min, glycolysis level (3.54±1.02) mpH/min, glycolytic capacity (5.94±0.47) mpH/min, and glycolytic reserve(2.40±0.61) mpH/min in the BaP-treated group were not statistically different from those in the vehicle control group (0.73±0.27) pmol/min, (2.71±0.66) mpH/min, (5.75±0.65) mpH/min, and (3.04±0.19) mpH/min, respectively (P > 0.05). The cell viabilities (90.2%±1.1%, 91.9%±3.1%, and 98.2%±2.1%) and SOD activities (67.28±0.43), (66.23±0.70), and (65.47±1.17) U/mg in the groups treated with vitamin E, vitamin C, and taurine in addition to BaP were significantly higher than those in the BaP-treated group84.2%±1.2% and (60.19±1.02) U/mg, while the ROS levels (34.53%±1.96%, 37.07%±2.42%, and 38.77%±2.31%) and MDA contents (0.477±0.095), (0.544±1.09), and (0.558±0.152) μmol/g were lower60.73%±3.15% and (0.837±0.104) μmol/g (P < 0.05). The basal OCR (4.30±0.36), (4.44±0.35), and (4.42±0.51) pmol/min, ATP-linked OCR (3.45±0.20), (3.70±0.19), and (3.60±0.34) pmol/min, and maximal respiration (5.53±0.03), (5.52±0.04), and (5.56±0.02) pmol/min in the groups treated with vitamin E, vitamin C, and taurine in addition to BaP were significantly higher than those in the BaP-treated group (2.64±0.43), (2.09±0.32), and (3.42±0.03) pmol/min, respectively (P < 0.05), and there was no significant difference in the spare respiratory capacity between the three groups (1.23±0.35), (1.08±0.31), and (1.14±0.50) pmol/min and the Bap-treated group (0.78±0.44) pmol/min (P > 0.05). The results of glycolysis stress test showed no differences in the three glycolysis function indicators among the five groups (P > 0.05).
Conclusion Exposure to BaP can cause oxidative damage in SH-SY5Y cells and reduce mitochondrial respiratory, which may be protected by vitamin E, vitamin C, and taurine.
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