氟化钠诱导成骨细胞内质网应激特征

Fluoride-Induced Endoplasmic Reticulum Stress in Osteoblasts

目的观察内质网应激在染氟成骨细胞中的特征,探索氟能否直接引起成骨细胞内质网应激。方法 2.5、5、10、20、40、80mg/L氟化钠染毒原代培养成骨细胞,并设立空白对照组和衣霉素阳性对照组。24h后Western blot检测成骨细胞内重链结合蛋白(heavy-chain binding protein,BIP)、X盒结合蛋白-1(X box binging protein1,XBP-1)、生长抑制DNA损伤基因(CCAAT/enhancer-binding protein-homologous protein,CHOP,也称GADD153)、蛋白质二硫键异构酶(protein disulfideismoerase,PDI)表达水平。结果各染毒组PDI蛋白表达量与空白对照与比较差异无统计学意义,XBP-1从10mg/L氟化钠组起表达明显高于空白对照组,并随氟剂量表达量增加。从2.5mg/L氟化钠组起骨细胞中的BIP表达高于对照组(P<0.05);从10mg/L氟化钠组起,各氟染毒组的CHOP与β-actin的比值比对照组高(P<0.05)。结论氟能够引起成骨细胞内质网应激。

Objective To observe endoplasmic reticulum stress in the osteobtasts directly treated with fluoride. Methods Primary cultured osteoblasts were treated with NaF at the doses of 0, 2.5, 5, 10, 20, 40 and 80 mg/L respectively for 24 hours, the positive control group was treated with tunicamycin. With Western blot, the expressions of heavy-chain binding protein （BIP）, X box hinging protein 1 （XBP-1）, CCAAT/enhancer-binding protein-homologous protein （CHOP） and protein disulfide isomerase （PDI） in the osteoblasts was determined. Results Compared with the control, no significant difference （P〉0.05） was observed in the expression of PDI in fluoride treated groups, and the expression of XBP-1 in 10-80 mg/L groups was significantly higher with a dose-dependent manner, and in 2.5-80 mg/L groups, the expression of BIP/β-actin was up-regulated （P〈0.05）, and in 10- 80 mg/L groups, the expression of CHOP/B-ACTIN was up-regulated （P〈0.05）. Conclusion Fluoride can indirectly induce endoplasmic reticulum stress in the osteoblasts.