The effect of fluoride on the expression of GRP-78 and caspase-12 in rat ameloblast

Abstract

Abstract: PURPOSE: To study the effect of GRP-78 and caspase-12 on fluoride-induced endoplasmic reticulum stress and apoptosis in rat ameloblast, and explore whether fluoride-induced endoplasmic reticulum stress results in the occurrence of apoptosis. METHODS: The cell activity of ameloblast cultured in various concentrations of fluoride was measured by CCK8 and flow cytometry; Real-time RT-PCR and Western blot were used to analyze the endoplasmic reticulum chaperone GRP-78 and caspase-12 genes and the expression of related proteins. The data was analyzed with SPSS13.0 software package. RESULTS: With the increasing concentration of fluoride, the cell activity of rat ameloblasts decreased gradually, and flow cytometry also showed that the number of apoptosis gradually increased; Real-time RT-PCR and Western blot showed the expression of GRP-78 and caspase-12 increased while the fluoride concentration increased. CONCLUSIONS: Excessive fluoride induces endoplasmic reticulum stress of rat ameloblast, and leads to cell apoptosis.

Key words: Fluoride, Ameloblast, Endoplasmic reticulum stress, Apoptosis

CLC Number:

R780.1

LI Jian,ZHANG Ying,MA Lin,ZHANG Kai-qiang,LEI Shuang,GU He-feng. The effect of fluoride on the expression of GRP-78 and caspase-12 in rat ameloblast[J]. Shanghai Journal of Stomatology, 2015, 24(1): 1-5.

References

[1] Wang X, Eno CO, Altman BJ, et al. ER stress modulates cellular metabolism [J]. Biochem J, 2011, 435(1): 285-296.
[2] Kubota K, Lee DH, Tsuchiya M, et al. Fluoride induces endoplasmic reticulum stress in ameloblasts responsible for dental enamel formation [J]. Biochem J, 2005, 280(24): 23197-23202.
[3] Holland RI. Fluoride inhibition of protein and DNA synthesis in cells in vitro [J]. Acta Pharmacol Toxicol (Copenh), 1979, 45(2): 96-101.
[4] Zhou R, Zaki AE, Eisenmann DR. Morphometry and autoradiography of altered rat enamel protein processing due to chronic exposure to fluoride [J]. Arch Oral Biol, 1996, 41(8-9): 739-747.
[5] He P, Zhang Y, Kim SO, et al. Ameloblast differentiation in the human developing tooth: effects of extracellular matrices [J]. Matrix Biol, 2010, 29(5): 411-419.
[6] Adamek E, Pawowska-Góral K, Bober K. In vitro and in vivo effects of fluoride ions on enzyme activity [J]. Ann Acad Med Stetin, 2005, 51(2): 69-85.
[7] Whitford GM. Determinants and mechanisms of enamel fluorosis[J]. Ciba Found Symp, 1997, 205: 226-241.
[8] 张凯强, 张颖, 刘璐, 等. 氟对大鼠切牙成釉细胞内质网伴侣分子表达的影响 [J]. 上海口腔医学, 2013, 22(5): 481-486.
[9] Yan Q, Zhang Y, Li W, et al. Mcromolar fluoride alters ameloblast lineage cells in vitro [J]. J Dent Res, 2007, 86(4): 336-340.
[10] Pfaffenbach KT, Lee AS. The critical role of GRP-78 in physiologic and pathologic stress [J]. Curr Opin Cell Biol, 2011, 23(2): 150-156.
[11] 马林, 张颖, 张凯强, 等. 内质网应激诱导细胞凋亡的机制及其在氟斑牙形成中的作用 [J]. 中国实用口腔科杂志, 2013, 6(6): 379-382.
[12] Zhang Y, Liu R, Ni M, et al. Cell surface relocalization of the endoplasmic reticulum chaperone and unfolded protein response regu-lator GRP-78/BIP [J]. J Biol Chem, 2010, 285(20): 150-165.
[13] Zwekh JE, Morris AC, Padelino PK, et al. Fluoride rapidly and transiently raises intracellular calcium in human osteoblasts [J]. J Bone Miner Res, 1990, 5(Suppl 1): S131-136.
[14] Nakagawa T, Zhu H, Morishima N, et al. Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta [J]. Nature, 2000, 403(6765): 98-103.
[15] Kim EM, Shin EJ, Choi JH, et al. Matrix metalloproteinase-3 is increased and participates in neuronal apoptotic signaling downstream of caspase-12 during endoplasmic reticulum stress [J]. J Biol Chem, 2010, 285(22): 16444-16452.