氟对大鼠成骨细胞骨保护蛋白表达的影响

目的 观察氟对体外培养大鼠乳鼠成骨细胞骨保护蛋白(osteoprotegerin,OPG)mRNA和蛋白表达的影响.方法 体外培养Wistar大鼠乳鼠成骨细胞,按染氟剂量分为0(对照)、1、2、4 mg/L组.分别在染氟48、72 h后提取RNA,采用反转录聚合酶链反应(RT-PCR)法检测OPG mRNA表达;采用酶连免疫吸附实验(ELISA)法检测培养液上清中的OPG蛋白表达.结果 大鼠乳鼠成骨细胞在体外染氟培养48、72 h,OPG mRNA表达组间比较差异均有统计学意义(F值分别为333.48、808.34,P<0.05).在染氟48 h,1、2、4 mg/L组OPGmRNA表达(0.810±0.043、0.819±0.031、0.870±0.044)与对照组(0.800±0.040)比较,差异均有统计学意义(P<0.05);在染氟72 h,1、2、4 mg/L组(0.933±0.047、1.031±0.051、1.240±0.062)高于对照组(0.805±0.020),差异均有统计学意义(P<0.05);OPGmRNA表达,72 h均高于48 h,染氟剂量和染氟时间有交互作用(F=204.16,P<0.05).在染氟培养48、72 h,OPG蛋白表达组间比较差异均有统计学意义(F值分别为7.49、41.31,P<0.05);组间两两比较,仅4 mg/L组(0.228±0.014、0.277±0.048)与对照组(0.205±0.012、0.229±0.010)比较,差异有统计学意义(P<0.05);OPG蛋白表达虽然72 h均高于48 h,但染氟剂量和染氟时间无交互作用(F=1.21,P>0.05).结论 氟可促进成骨细胞OPG mRNA和蛋白表达,这种作用与染氟的剂量以及作用时间有关.

Influence of fluoride on osteoprotegerin mRNA and protein expression in rat osteoblasts: an in vitro study

Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of dosesO(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.