铝暴露对大鼠成骨细胞Toll样受体4表达的影响  

Effects of aluminum exposure on the expression of toll-like receptor 4 in osteoblast

作　　者：陈玉楼[1] 李兴海[1] 陈铁[1] 周毅[1] 裴梓峰[1] 

机构地区：[1]沈阳医学院沈洲医院骨外科,辽宁沈阳110032

出　　处：《中国现代医学杂志》2014年第5期28-32,共5页China Journal of Modern Medicine

基　　金：辽宁省教育厅科学技术研究项目(No:2008848)

摘　　要：目的通过观察toll样受体4(TLR4)在铝暴露大鼠成骨细胞中的表达,探讨铝的骨毒性分子机制。方法原代培养新生SD大鼠颅盖骨成骨细胞,分为对照组和铝暴露组,铝暴露组细胞用含有AlCl3(100μmol)的DMEM培养基培养,两组细胞分别培养0、12、24和48 h。Real-time RT-PCR方法检测各时间点成骨细胞中TLR4 mRNA表达;Western blotting检测各时间点成骨细胞TLR4蛋白的表达情况;MTT检测各组各时间点的细胞生存率情况。结果在12、24和48 h时间点,铝暴露各组细胞的TLR4 mRNA和蛋白表达明显高于对照组(P<0.05);在铝暴露各组中,与0 h组相比,TLR4 mRNA和蛋白表达随着时间的增加而增强(P<0.05);在12、24和48 h时间点,铝暴露各组细胞的生存率明显低于对照组;在铝暴露各组中,与0 h比较,成骨细胞的生存率随时间的增加而下降,呈时间依赖性(P<0.05)。结论铝暴露抑制成骨细胞生存率,可能与上调TLR4表达相关。[ Objective] To explore the molecular mechanisms of aluminum bone toxicity by observing the changes of toll-like receptor 4 expression in rat osteoblasts exposed to aluminum. [Methods] The calvarial osteoblast cells were obtained from the neonatal SD rats and cultured in DMEM containing 10% Fetal Bovine serum. The ceils were divided into two groups： the control and the aluminum-treated. The aluminum-treated was cultured in DMEM media containing A1C13 （100 t^mol） for 0 h, 12 h, 24 h and 48 h, respectively. Real- time RT-PCR and Western blotting methods were used to determine the expressions of TLR4 mRNA and protein. MTr assay was used to determine the survival rate of osteoblast cells at each time point. [Results] Compared with control groups, the expression levels of both TLR4 mRNA and protein in osteoblast cells were significantly increased at time points of 12 h, 24 h and 48 h （P 〈0.05）; and among the aluminum exposure groups, the expression levels of both TLR4 mRNA and protein were gradually increased, compared with 0 h, in a time dependence manner （P〈0.05）; At time points of 12 h, 24 h and 48 h, the survival rate of osteoblast cells exposed to aluminum was significantly decreased compare with that of control groups; and among the a- luminum exposure groups, the survival rate of the osteoblast cells was gradually decreased, compared with 0 h, in a time-dependent manner （P〈0.05）. [Conclusion] Aluminum exposure plays an inhibitory role on the osteoblast survival rate, which may be related to the enhancement in the expression of TLR4.

关 键 词：铝暴露 TOLL样受体4 成骨细胞 

分 类 号：Q26[生物学—细胞生物学]