Henan Provincial Open Laboratory of Key Disciplines, Environment and Animal Products Safety, College of Animal Science and Technology, Henan University of Science and Technology, Kaiyuan Avenue 263, Luoyang, Henan, 471000, PR China.
Henan Provincial Open Laboratory of Key Disciplines, Environment and Animal Products Safety, College of Animal Science and Technology, Henan University of Science and Technology, Kaiyuan Avenue 263, Luoyang, Henan, 471000, PR China.
Chemosphere. 2018 May;199:297-302. doi: 10.1016/j.chemosphere.2018.02.057. Epub 2018 Feb 9.
To investigate the mechanisms of fluoride-induced apoptosis, a fluoride-induced C2C12 skeletal muscle cell (C2C12 cell) model was established in this study, and the viability of the C2C12 cells was measured using an MTT assay. Cell morphological changes were observed via haematoxylin and eosin staining and transmission electron microscopy. Apoptosis was monitored through Hoechst staining. The mRNA and protein expression of PI3K, PDK1, AKT1, BAD, Bcl-2, Bax and caspase-9 were detected through real-time PCR and western blotting, respectively. The results showed that the survival rates of C2C12 cells decreased gradually with an increasing fluoride doses. The C2C12 cell structure was seriously damaged by fluoride, presenting with pyknosis, mitochondrial ridge disruption and swollen endoplasmic reticulum. Furthermore, the expression of mRNA in PI3K, BAD, Bcl-2, Bax and caspase-9 were significantly increased in the fluoride group (P < 0.01), while the expression of PDK1 was markedly decreased (P < 0.01). The expression of protein in BAD, Bcl-2 and Bax were significantly increased in the fluoride group (P < 0.01), while the expression of PDK1 and P-AKT1 was markedly decreased (P < 0.01). In conclusion, fluoride-induced apoptosis in C2C12 cells is related to the PI3K/AKT signaling pathway.
为了研究氟化物诱导细胞凋亡的机制，本研究建立了氟化物诱导的 C2C12 骨骼肌细胞（C2C12 细胞）模型，并通过 MTT 测定法测量 C2C12 细胞的活力。通过苏木精-伊红染色和透射电子显微镜观察细胞形态变化。通过 Hoechst 染色监测细胞凋亡。通过实时 PCR 和 Western blot 分别检测 PI3K、PDK1、AKT1、BAD、Bcl-2、Bax 和 caspase-9 的 mRNA 和蛋白表达。结果表明，随着氟化物剂量的增加，C2C12 细胞的存活率逐渐下降。氟化物严重破坏 C2C12 细胞结构，出现核固缩、线粒体嵴断裂和内质网肿胀。此外，氟化物组中 PI3K、BAD、Bcl-2、Bax 和 caspase-9 的 mRNA 表达显著增加（P<0.01），而 PDK1 的表达明显降低（P<0.01）。氟化物组中 BAD、Bcl-2 和 Bax 的蛋白表达明显增加（P<0.01），而 PDK1 和 P-AKT1 的表达明显降低（P<0.01）。总之，氟化物诱导 C2C12 细胞凋亡与 PI3K/AKT 信号通路有关。
