Deng Huidan, Kuang Ping, Cui Hengmin, Chen Lian, Fang Jing, Zuo Zhicai, Deng Junliang, Wang Xun, Zhao Ling
College of Veterinary Medicine, Sichuan Agricultural University, Ya'an, China.
Key Laboratory of Animal Diseases and Environmental Hazards of Sichuan Province, Ya'an, China.
Oncotarget. 2016 Oct 18;7(42):67880-67900. doi: 10.18632/oncotarget.12081.
Though fluorine has been shown to induce apoptosis in immune organs in vivo, there has no report on fluoride-induced apoptosis in the cultured lymphocytes. Therefore, this study was conducted with objective of investigating apoptosis induced by sodium fluoride (NaF) and the mechanism behind that in the cultured splenic lymphocytes by flow cytometry, western blot and Hoechst 33258 staining. The splenic lymphocytes were isolated from 3 weeks old male ICR mice and exposed to NaF (0, 100, 200, and 400 μmol/L) in vitro for 24 and 48 h. When compared to control group, flow cytometry assay and Hoechst 33258 staining showed that NaF induced lymphocytes apoptosis, which was promoted by decrease of mitochondria transmembrane potential, up-regulation of Bax, Bak, Fas, FasL, caspase 9, caspase 8, caspase 7, caspase 6 and caspase 3 protein expression (P < 0.05 or P <0.01), and down-regulation of Bcl-2 and Bcl-xL protein expression (P <0.05 or P <0.01). The above-mentioned data suggested that NaF-induced apoptosis in splenic lymphocytes could be mediated by mitochondrial and death receptor pathways.
尽管已有研究表明氟在体内可诱导免疫器官细胞凋亡,但关于氟化物诱导培养淋巴细胞凋亡的报道尚未见。因此,本研究采用流式细胞术、蛋白质免疫印迹法和Hoechst 33258染色法,旨在探讨氟化钠(NaF)诱导培养的脾淋巴细胞凋亡及其机制。从3周龄雄性ICR小鼠分离脾淋巴细胞,体外分别用0、100、200和400 μmol/L NaF处理24 h和48 h。与对照组相比,流式细胞术检测和Hoechst 33258染色结果显示,NaF可诱导淋巴细胞凋亡,其机制可能与线粒体跨膜电位降低、Bax、Bak、Fas、FasL、caspase 9、caspase 8、caspase 7、caspase 6和caspase 3蛋白表达上调(P < 0.05或P <0.01)以及Bcl-2和Bcl-xL蛋白表达下调(P < 0.05或P <0.01)有关。上述结果提示,NaF诱导的脾淋巴细胞凋亡可能由线粒体途径和死亡受体途径介导。