Fas/FasL途径在氟致人神经母细胞瘤SH-SY5Y细胞凋亡中的作用

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、80 mg/L].SH-SY5Y细胞进行染毒,24 h后检测细胞存活率、凋亡率和Fas、FasL mRNA表达;选择40 mg/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、80 mg/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、80 mg/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(Jp<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)、80 mg/L组(0.99±0.12)]和FasL/β-actin[40 mg/L组(0.96±0.42)、80 mg/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(,值分别为32.89、18.46、14.69,P<0.01),NaF与ZB4之间存在拈抗作用(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.

Effect of Fas/FasL pathway on fluoride-induced apoptosis in human neuroblastoma SH-SY5Y cells

Objective To explore the effect of Fas/FasL pathway on fluoride.induced apoptosis in hurnan neumbla8toma SH-SY5Y 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 O(control),20,40,80 mg/L sodium nuoride(NaF)for 24 hours/n 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 activaling or neutralizing anti-Fas antibody(CH11 or ZB4)also observed respectively.Results Compared with the control group(100.00%), the cell surval rates in 40,80 mg/L NaF-treated groups[(84.63±2.57)%,(69.04±5.63)%]were significandy 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 thal 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 ms/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[Eas/β-actin(0.50±0.33),FasL/β-actin(0.58±0.23)],both the difference had 8tatistical significances (P<0.05).NaF and CH I 1 had a synergisfic effect on apoptosis and mRNA expression levels of Fas and FasLL(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-Y5Y cells,and the Fas/FasL pmhway may play an important role in NaF-induced apoptosis.