College of Chemistry and Life Science at Zhejiang Normal University, Jinhua, Zhejiang, 321004, PR China.
College of Sports and Health Science, Zhejiang Normal University, Jinhua, Zhejiang, 321004, PR China.
Environ Toxicol Pharmacol. 2017 Sep;54:74-79. doi: 10.1016/j.etap.2017.06.018. Epub 2017 Jun 30.
This study investigated the effects of fluoride exposure on the mRNA expression of Cav1.2 calcium signaling pathway and apoptosis regulatory molecules in PC12 cells. The viability of PC12 cell receiving high fluoride (5.0mM) and low fluoride (0.5mM) alone or fluoride combined with L-type calcium channel (LTCC) agonist/inhibitor (5umol/L FPL6417/2umol/L nifedipine) was detected using cell counting kit-8 at different time points (2, 4, 6, 8, 12, 10, and 24h). Changes in the cell configuration were observed after exposing the cells to fluoride for 24h. The expression levels of molecules related to the LTCC were examined, particularly, Cav1.2, c-fos, CAMK II, Bax, and Bcl-2. Fluoride poisoning induced severe cell injuries, such as decreased PC12 cell activity, enhanced cell apoptosis, high c-fos, CAMKII, and Bax mRNA expression levels. Bcl-2 expression level was also reduced. Meanwhile, high fluoride, high fluoride with FPL64176, and low fluoride with FPL64176 enhanced the Cav1.2 expression level. In contrast, low fluoride, high fluoride with nifedipine, and low fluoride with nifedipine reduced the Cav1.2 expression level. Thus, Cav1.2 may be an important molecular target for the fluorosis treatment, and the LTCC inhibitor nifedipine may be an effective drug for fluorosis.
本研究探讨了氟暴露对 PC12 细胞 Cav1.2 钙信号通路和细胞凋亡调控分子 mRNA 表达的影响。分别用细胞计数试剂盒-8 在不同时间点(2、4、6、8、12、10 和 24h)检测单独接受高氟(5.0mM)和低氟(0.5mM)以及氟化物与 L 型钙通道(LTCC)激动剂/抑制剂(5μmol/L FPL6417/2μmol/L 硝苯地平)的 PC12 细胞的活力。暴露于氟化物 24h 后观察细胞形态的变化。检测与 LTCC 相关的分子的表达水平,特别是 Cav1.2、c-fos、CAMK II、Bax 和 Bcl-2。氟中毒导致严重的细胞损伤,如 PC12 细胞活力下降,细胞凋亡增加,c-fos、CAMKII 和 Bax mRNA 表达水平升高,Bcl-2 表达水平降低。同时,高氟、高氟加 FPL64176 和低氟加 FPL64176 增强了 Cav1.2 的表达。相反,低氟、高氟加硝苯地平以及低氟加硝苯地平降低了 Cav1.2 的表达水平。因此,Cav1.2 可能是氟中毒治疗的重要分子靶点,LTCC 抑制剂硝苯地平可能是一种有效的氟中毒治疗药物。