摘要
目的观察氟通过胞外信号调控激酶（ERK）通路对成骨细胞（MC3T3-E1）增殖作用的影响。方法取小鼠成骨细胞（MC3T3-E1）进行体外培养，加入不同浓度的氟（F-，0、200、400、600、1000、2000、4000、8000、10000μmol／L）培养24、48h，甲基噻唑蓝（MTr）法筛选出促进细胞增殖的最适浓度。根据最适浓度，将成骨细胞单纯随机分为3组：对照组（F-，0μmol／L）、染氟组（F-，400／μmol／L）、染氟抑制组（F-，400μmol／L＋PD91805，10μmmol／L）。培养48h后应用流式细胞术检测各组细胞周期；Westernbolt法和免疫荧光法检测各组磷酸化ERK（P-ERK）蛋白表达。结果400Ixmol／L的氟是促进成骨细胞增殖的最适合浓度。与对照组比较[（76．12±10．08）％、（2．064-0．31）％]，染氟组G0／G1期细胞数[（63．044-8．12）％]明显减少，S期细胞数[（9．13±2．08）％]明显增多（P均〈0．05）；而染氟抑制组G0／G1期细胞数[（92．11±9．01）％]明显增多（P〈0．05）。Westernblot结果表明，与对照组[（100．00±0．00）％]比较，染氟组成骨细胞P-ERK蛋白表达水平[（131．244-13．88）％]明显增高（P〈0．05），染氟抑制组P-ERK蛋白表达[（91．33±9．68）％]未见明显改变（P〉0．05）；免疫荧光法检测结果与Westernblot法相似。结论400txmol／L氟可以促进成骨细胞增殖，ERK通路在氟促进成骨细胞的增殖作用中起到了关键的作用。
Objective To study the effect of fluorine on proliferation of osteoblast through extra cellular signal-regulated protein kinase （ERK） signaling pathway. Methods Mouse osteoblasts （MC3T3-E 1 ） were cultured in vitro with different concentrations of fluoride for 24 and 48 h （the concentrations of F- were 0, 200, 400, 600, 1000, 2000, 4000, 8000, 10 000 p, mol/L, respectively）. The optimum concentration for promotion of cell proliferation was determined by methyhhiophene tetrazolium （MTF） assay. According to the optimum concentration, the cells were randomly divided into three groups： control group （0 μmol/L F-）; fluorine group （400 μmol/L F-）; fluorine and MAPK inhibitor PD98059 group（400 μmol/L F± 10 p, mmol/L PD98059）. Cell cycle was detected by flow cytometry after 48 h culture. The expression of PERK protein was determined by Western blotting and immunofluorescence. Results The optimum concentration of fluorine for proliferation of osteoblasts was 400 txmol/L. Compared with the control group[ （76.12 ± 10.08）%, （2.06 ± 0.31）%], the number of cells in G0/G1 phase[ （63.04 ± 8.12）%] reduced and the number of cells in S phase [ （9.13 ± 2.08）% ] increased in fluorine group （all P 〈 0.05 ） ; but the number of cells in G0/G1 phase [ （92.11 ± 9.01）% ] in fluorine and mitogen-activated protein kinases（MAPK） inhibitor PD98059 group was significantly increased（P 〈 0.05）. Western blotting results showed that： compared with the control group[ （100.00 ± 0.00）%], the expression of P-ERK protein in fluorine group[ （131.24 ± 13.88）%] was significantly higher（P 〈 0.05 ）, but the expression of P-ERK protein in fluorine and MAPK inhibitor PD98059 group [ （91.33± 9.68 ）% ] was not significantly changed（P 〉 0.05 ）. The results of immunofluorescence were similar to that of Western blotting. Conclusions Fluorine at the concentration of 400 μmol/L can promote the proliferation ofosteoblasts. ERK signaling pathway has played a key role in the proliferation of osteoblasts.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2012年第2期140-143,共4页
Chinese Jouranl of Endemiology
基金
国家自然科学基金（81170808、30771813）
关键词
成骨细胞
氟
细胞增殖
细胞外信号调节MAP激酶类
Osteoblasts
Fluorine
Cell proliferation
Extracellular signal-regulated MAP kinases
