目的探讨Fas/FasL途径在氟致人神经母细胞瘤SH—SY5Y细胞凋亡中的作用。方法用不同剂量氟化钠[NaF,0(对照)、20、40、80mg/L]对SH—SY5Y细胞进行染毒,24h后检测细胞存活率、凋亡率和Fas、FasL mRNA表达;选择40mg/L NaF组,观察在Fas受体激动剂(CH11)或拮抗剂(ZB4)作用下细胞凋亡率及Fas、FasL mRNA表达水平的改变。结果40、80mg/L组细胞存活率[(84.63±2.57)%、(69.04±5.63)%]明显低于对照组(100.00%),组间比较差异有统计学意义(P〈0.01);细胞凋亡率随染毒剂量的升高呈上升趋势,40、80mg/L组细胞凋亡率[(8.54±1.95)%、(17.94±2.71)%]明显高于对照组[(3.32±1.33)%],组间比较差异有统计学意义(P〈0.05);NaF能不同程度地上调Fas、FasL mRNA表达,使Fas/β-actin[40mg/L组(0.94±0.51)、80mg/L组(0.99±0,12)]和Fast/β-actin[40mg/L组(0.96±0.42)、80mg/L组(0.99±0.24)]比值增加,与对照组[Fas/β-actin(0.50±0.33)、FasL/β-actin(0.58±0.23)]比较,差异均有统计学意义(P〈0.05)。在对细胞凋亡率和Fas、FasL mRNA表达水平的影响中,NaF与CH11之间存在协同作用(F值分别为32.89、18.46、14.69,P〈0.01),NaF与ZB4之间存在拮抗作用(F值分别为5.73、24.26、10.17,P〈0.05或〈0.01)。结论NaF可诱导SH—SY5Y细胞凋亡,Fas/FasL途径在NaF诱导SH—SY5Y细胞凋亡中起重要作用。
Objective To explore the effect of Fas/FasL pathway on fluoride-induced apoptosis in human neuroblastoma SH-SYSY cells. Methods The cell survival rate, percentage of apoptosis, and mRNA expression levels of Fas and FasL were measured respectively after the SH-SY5Y cells were exposed to 0 (control),20,40,80 mg/L sodium fluoride (NaF) for 24 hours in vitro. Furthermore, the changes of the percentage of apoptosis and mRNA expression levels of Fas and FasL in 40 mg/L NaF-treated groups incubated with activating or neutralizing anti-Fas antibody(CHll or ZB4) also observed respectively. Results Compared with the control group(100.00%), the cell survival rates in 40,80 mg/L NaF-treated groups[ (84.63 ± 2.57)%, (69.04 ± 5.63)%] were significantly lower(P 〈 0.01 ). The percentage of apoptosis in 40,80 mg/L NaF-treated groups[(8.54 ± 1.95)%, (17.94 ± 2.71 )%] were higher(P 〈 0.05) than that in the control group[ (3.32 ± 1.33)%], and increased with the dose of NaF. NaF could up-regulate Fas and FasL mRNA expression, and increased the Fas/β-actin [40 mg/L group (0.94 ± 0.51 ), 80 mg/L group(0.99 ± 0.12)] and FasL/β-actin[40 mg/L group(0.96 ± 0.42), 80 mg/L group(0.99 ± 0.24)] ratio, compared with the control [Fas/β-actin (0.50 ± 0.33), FasL/β-actin(0.58 ± 0.23)], both the difference had statistical significances (P 〈 0.05). NaF and CH11 had a synergistic effect on apoptosis and mRNA expression levels of Fas and FasL(F = 32.89,18.46,14.69, P 〈 0.01 ) while NaF and ZB4 had an antagonistic effect (F = 5.73,24.26, 10.17, P 〈 0.05 or 〈 0.01 ). Conclusion NaF exposure can cause apoptosis in SH-SY5Y cells, and the Fas/FasL pathway may play an important role in NaF-induced apoptosis.