摘要
目的研究唑来膦酸(ZOL)对破骨细胞黏附以及整合素αv和β3基因表达的影响。方法体外诱导小鼠RAW264.7细胞向破骨细胞分化,通过抗酒石酸酸性磷酸酶(TRAP)染色及牙本质吸收陷窝检测以评价破骨细胞生成情况。将细胞分为对照组及ZOL处理组两组,后者用1×10-6 mol·L-1的ZOL处理2 d,用结晶紫染色法检测细胞黏附情况,用实时荧光定量聚合酶链反应、Western blot和免疫荧光化学法检测整合素αv、β3 m RNA及蛋白表达水平。结果 TRAP染色及牙本质吸收陷窝检测提示有多核破骨细胞生成。ZOL处理组破骨细胞黏附能力较对照组显著降低(P<0.01)。ZOL处理组整合素αv、β3 m RNA水平分别为0.66±0.05、0.59±0.08,显著低于对照组的1.01±0.01和1.01±0.02(P<0.01);蛋白表达水平分别为31 934.84±112.91、18 812.79±194.13,较对照组(52 517.81±211.72、31 441.93±456.87)分别下降了39.19%和40.17%(P<0.01)。免疫荧光化学检测显示,ZOL处理使整合素αv、β3荧光强度(9.491±0.748、4.744±0.759)较对照组(15.159±1.143、11.418±1.095)分别降低了37.39%和58.45%(P<0.01)。结论 ZOL可抑制破骨细胞黏附并下调整合素αv、β3表达;ZOL的上述作用可能参与对破骨细胞性骨吸收的抑制。
Objective To explore the effect of zoledronate（ZOL） on the osteoclast adhesion and expression of integrin αv and β3 in vitro.Methods Mice RAW264.7 cells were used for osteoclast differentiation in vitro,and osteoclastogenesis was examined by tartrate-resistant acid phosphatase（TRAP） staining and dentin resorption lacunae examination.The cells were then divided into 2 groups,the control group and ZOL treatment group（treated with 1×10-6 mol·L-1 ZOL for 2 d）.The adhesion ability of osteoclasts and m RNA and the protein expressions of integrin αv and β3 were examined by crystal violet staining,real-time fluorescence quantitative polymerase chain reaction,Western blot analysis,and immunofluorescent chemistry.Results TRAP staining and dentin resorption lacunae examination revealed the formation of multi-nuclear osteoclasts.ZOL treatment significantly decreased the adhesion ability of osteoclasts（P〈0.01）.In the ZOL-treated group,the m RNA levels of integrin αv and β3 were 0.66±0.05 and 0.59±0.08,respectively.In the control group,the m RNA levels of integrin αv and β3 were 1.01±0.01 and 1.01±0.02,respectively;these values were higher than those in the ZOL-treated group（P〈0.01）.The protein level of integrin αv and β3 in the ZOL-treated group（31 934.84±112.91 and 18 812.79±194.13） was downregulated by approximately 39.19% and 40.17%,respectively,compared with those in the control group（52 517.81±211.72 and 31 441.93±456.87）（P〈0.01）.Immunofluorescent examination showed that the fluorescent intensities of integrin αv and β3 in the ZOL-treated group（9.491±0.748 and 4.744±0.759） were also significantly decreased compared with those in the control group（15.159±1.143 and 11.418±1.095）（P〈0.01）.Conclusion ZOL significantly inhibits osteoclast adhesion and downregulates integrin αv and β3 expression,thus contributing to the ZOL-induced inhibition of osteoclastmediated bone resorption.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2014年第6期547-551,共5页
West China Journal of Stomatology
基金
国家自然科学基金资助项目(81270965,81101448)
河北省自然科学基金资助项目(C2011401044)
