Relationship between intracellular calcium and reactive oxygen species in sodium fluoride-induced injury in human neuroblastoma SH-SY5Y cells
10.3760/cma.j.issn.1000-4955.2009.02.003
- VernacularTitle:细胞内钙离子和活性氧在氟化钠致人神经母细胞瘤SH-SY5Y细胞损伤中的关系
- Author:
Zhi-xia, XU
;
Ba-yi, XU
;
Tao, XIA
;
Ping, HE
;
Ping, GAO
;
Li-juan, GUO
;
Qiang, NIU
;
Nan, HUNAG
;
Ai-guo, WANG
- Publication Type:Journal Article
- Keywords:
Sodium fluoride;
Neuroblastoma;
Cell line;
Reactive oxygen species;
Calcium ion
- From:Chinese Journal of Endemiology
2009;28(2):126-129
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the relationship between intracellular calcium levels ([Ca2+]1) and reactive oxygen species (ROS) in sodium fluoride (NaF)-induced injury in human neuroblastoma SH-SY5Y cells. Methods The levels of [Ca2+]1 and ROS were measured in different exposed times(0,3,6,12,18,24 h) respectively after SH-SY5Y cells were exposed to 40 mg/L NaF in vitro, and the optimal expose time was selected. Furthermore, the changes of [Ca2+]1, ROS and LDH levels in 40 mg/L NaF-treated groups incubated with 38.23 mg/L BAPTA-AM or 380.40 mg/L ethylene glycol-bis-(beta-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA) or 16.32 mg/L N-acetyl-L-cysteine(NAC) also observed at the optimal expose time(12 h), respectively. Results At 3,6,12,18 and 24 h, [Ca2+]1 level(5620.0±226.3,4775.5±85.6,3312.3±87.5, 3047.0±75.0,2717.0±66.5) was significantly increased, and so was the ROS level(4449.53±324.61,7463.07±117.43,20 227.33±178.04,8817.56±200.13, 7975.61±92.90) except at 3 h, compared with 0 h(2115.0±24.0,4098.01±21.22, all P<0.05). The levels of [Ca2+]1 and ROS reached the peak at 3 h and 12 h, respectively. [Ca2+]1 and LDH levels in NaF-treated group [3279.5±94.0, (1057.50±64.35)U/L], NaF+NAC treated group[ 3583.0±350.7, (561.02±85.50)U/L], NaF+EGTA treated groups[3701.5±157.7, (1074.50±86.97)U/L], and BAPTA-AM treated group[2766.5±38.9, (521.43±40.80)U/L] had increased, compared with the control[2022.5±118.1, (186.97±8.73)U/L], the difference being statistically significant (P<0.05). ROS levels in NaF-treated group (19 003.04±332.34), and NaF+EGTA treated group(19 170.12±95.46) was higher than that in the controls(4060.98±145.66), the difference being statistically significant (P<0.05). NaF and NAC had antagonistic effect on ROS and LDH levels (F=976.11,43.54,P<0.05). And NaF and BAPTA-AM had antagonistic effect on [Ca2+]1, ROS and LDH levels (F=15.65,1515.53,115.00, P<0.05). Conclusions NaF-related calcium is released from the site of intracellular calcium storage, which induces ROS production, both of them caused cytotoxicity and the increase of LDH level in human neuroblastoma SH-SY5Y cells.
