过量氟对体外培养LS8细胞中炎症因子表达的影响

Effect of high fluoride on inflammation cytokines in LS8 cells

目的观察过量氟对小鼠成釉细胞样细胞(LS8细胞)中炎症因子表达的影响。方法取对数生长期LS8细胞进行培养,待细胞密度达到70%-80%时,加入含有2 mmol/L NaF的培养基分别培养0,12,24,48 h,收集细胞,qRT-PCR检测白细胞介素1β(interleukin 1β,IL-1β)、IL-6、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)及核因子κB(nuclear factor-kappa B,NF-κB)mRNA水平。另外,加入含有2 mmol/L NaF的培养基分别培养0,8,15,30,60,120 min,收集细胞,Western blotting检测p-P38、P38蛋白表达。结果NaF干预体外培养的LS8细胞后,IL-1β、TNF-α在培养24,48 h时,IL-6在培养48 h时,及NF-κB在培养12,24 h时mRNA表达均显著增加(P<0.05)。NaF干预LS8细胞30,60,120 min后,P38磷酸化显著增加(P<0.05)。结论过量氟可导致LS8细胞炎症反应发生,NF-κB及p38信号通路可能参与其中。

Objective To investigate the effect of high fluoride on the inflammation of ameloblast-like cells(LS8 cells).Methods LS8 cells were chosen and cultured.When the cells reached 70%-80%,they were incubated with medium containing 2 mmol/L NaF for 0,12,24,48 h,and then the mRNA levels of IL-1β,IL-6,TNF-αand NF-κB in LS8 cells were determined by qRT-PCR.Meanwhile,LS8 cells were incubated with medium containing 2 mmol/L NaF for 0,8,15,30,60,120 min,and then the protein levels of p-P38 and P38 in LS8 cells were assessed by Western blotting.Results The mRNA levels of IL-1βand TNF-αat 24,48 h,IL-6 at 48 h and NF-κB at 12,24 h were increased in LS8 cells after treated with NaF in vitro(P<0.05).The expression of p-P38 was significantly increased in LS8 cells with 2 mmol/L NaF for 30,60,120 min(P<0.05).Conclusion High fluoride exposure may cause the inflammation in LS8 cells in vitro via NF-κB and p38 signal pathway.