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目的探讨结核分枝杆菌超声裂解产物(Mt sonicate)体外诱导小鼠破骨细胞的形成及OPG/RANKL表达变化与裂解物的浓度、时间的依赖性,研究结核杆菌引起骨破坏的相关机制。方法原代培养的BALB/c小鼠骨髓基质细胞以高、中、低浓度的Mt sonicate进行干预,在1、3、7、14d进行TRAP染色,观察破骨细胞的形成,并采用Realtime RT-PCR检测各浓度、时间的骨保护素(OPG)/核因子Kappa B受体活化因子配基(RANKL)mRNA表达。结果 Mt sonicate在体外可诱导小鼠骨髓基质细胞向破骨细胞的转化;破骨细胞数量与Mt sonicate浓度在3、7d时呈显著的正相关(3d:r=0.417,P<0.05;7d:r=0.463,P<0.05);3个浓度干预下,破骨细胞数量在1～7d逐渐增多,7～14d则逐渐减少。7d时,高、中、低浓度Mt sonicate干预下OPG/RANKL mRNA表达比分别为(1.35±0.11)、(7.38±1.15)、(9.85±1.26)。结论小鼠骨髓基质细胞向破骨细胞转化的实验中,存在结核分枝杆菌裂解物的浓度和时间依赖性,且与OPG/RANKL mRNA的表达比呈负相关,破骨细胞形成时的数量与活性受到结核分枝杆菌的调控,其分子机制与OPG/RANKL系统的变化密切相关。
Abstract：Objective This paper aims to explore the formation of broken bone cell in vitro mouse bone marrow stromal cells by Mycobacterium tuberculosis ultrasound pyrolysis products(Mt sonicate)induction,the lysate concentration dependence of the time,and the mechanisms of bone destruction caused by TB bacilli.Methods Primary cultures of BALB/c mice bone marrow stromal cells were intervened with high,medium and low concentrations of Mt sonicate on 1,3,7,14d;osteoclast formation was observed with TRAP staining and Realtime RT-PCR to detect the concentration and time of the OPG/RANKL mRNA expression.Results Mt sonicate can induce in vitro transformation of mouse bone marrow stromal cells into osteoclasts;the number of osteoclasts and Mt sonicate concentration on 3d and 7d showed a significant positive correlation(3d:r=0.417 P<0.05;7d:r=0.463 P<0.05);with three concentrations intervention,the number of osteoclasts gradually increased from 1d to 7d,gradually reduced from 7to14d;on 7d,OPG/RANKL mRNA expression ratios in each group of Mt sonicate intervened cells in high medium and low concentrations were 1.35±0.11,7.38±1.15,9.85±1.26.Conclusion There exist the dependence of Mycobacterium tuberculosis lysate concentration and time in the experiments on mouse bone marrow stromal cells into osteoclasts transformation,and the ratio of OPG/RANKL mRNA expression showed a negative relevance;the quantity and activity of osteoclastogenesis was regulated by the Mycobacterium tuberculosis;the molecular mechanisms of changes are closely related with OPG/RANKL system.
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基本信息:
中图分类号:R580
引用信息:
[1]张元豫,张浩沙强,李坤.结核分枝杆菌裂解物诱导破骨细胞及OPG/RANKL表达变化实验研究[J].新疆医科大学学报,2013,36(10):1437-1441.
基金信息:
新疆维吾尔自治区自然科学基金(2011211A054)
2013-10-15
2013-10-15