PI3K/AKT signaling pathway involvement in fluoride-induced apoptosis in C2C12 cells.
PI3K/AKT signaling pathway involvement in fluoride-induced apoptosis in C2C12 cells.
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DOI：
10.1016/j.chemosphere.2018.02.057
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发表时间：
2018-05
期刊：
影响因子：
8.8
通讯作者：
Bian-hua Zhou;Pan-pan Tan;L. Jia;Wen-peng Zhao;Ji-cang Wang;Hong-wei Wang
中科院分区：
文献类型：
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作者：
Bian-hua Zhou;Pan-pan Tan;L. Jia;Wen-peng Zhao;Ji-cang Wang;Hong-wei Wang
为探讨氟化物诱导细胞凋亡的机制，本研究建立氟化物诱导的C2C12骨骼肌细胞（C2C12 细胞）模型，采用MTT法测定C2C12 细胞的活力。通过苏木精、伊红染色和透射电镜观察细胞形态学变化。Hoechst染色检测细胞凋亡。采用real-time PCR和western blotting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，C2C12 细胞存活率随氟剂量的增加而逐渐降低。氟化物严重破坏C2C12 细胞结构，表现为固缩、线粒体脊断裂、内质网肿胀。氟组PI3K、BAD、Bcl-2、Bax、caspase-9 mRNA表达量显著升高（P< 0.01），PDK1 mRNA表达量显著降低（P< 0.01）。氟组BAD、Bcl-2、Bax蛋白表达量显著升高（P< 0.01），PDK1、P- akt1表达量显著降低（P< 0.01）。综上所述，氟化物诱导C2C12 细胞凋亡与PI3K/AKT信号通路有关。
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.