摘要
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、4mg／L组。分别在染氟48、72h后提取RNA，采用反转录聚合酶链反应（RT—PCR）法检测OPGmRNA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、72h，OPGmRNA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48h，1、2、4mg／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72h，1、2、4mg／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，72h均高于48h，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、72h，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4mg／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72h均高于48h，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OPGmRNA和蛋白表达，这种作用与染氟的剂量以及作用时间有关。
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 doses[0（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 ± 0.043, 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 rag/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 = 204.16, 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.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2009年第2期134-137,共4页
Chinese Jouranl of Endemiology
基金
国家自然科学基金（30471498）
关键词
骨保护蛋白
成骨细胞
氟化物
反转录聚合酶链反应
酶联免疫吸附测定
Osteoprotegerin
Osteoblast
Fluorides
Reverse transcription polymerase chain reaction
Enzyme-linked immunosorbent assay
