HMGB1对大鼠骨髓基质干细胞增殖和分化的影响

The effects of HMGB1 on the proliferation and differentiation of bone marrow stromal cells from rats

目的:探讨高迁移率族蛋白B1(HMGB1)对于大鼠骨髓基质干细胞BMSCs增殖、成骨/破骨的影响。方法:原代培养的大鼠BMSCs,分别以浓度为0,100,500 ng/mL HMGB1作用于BMSCs,MTT法检测细胞数目;以浓度为0,100,500 ng/mL HMGB1作用于BMSCs,7d后行ALP染色和RT-PCR检测成骨/破骨基因的表达(骨钙素OCN,骨保护素OPG,核因子κB受体活化因子配体RANKL)。结果:HMGB1在100、500 ng/mL浓度时,第3 d和第5 d BMSCs的OD值有明显升高,在第7 d对BMSCs的ALP染色没有明显的改变,OCN和OPG基因的表达也没有明显变化,HMGB1明显提高了BMSCs的RANKL及RANKL/OPG基因表达。结论:HMGB1对大鼠BMSCs的增殖和破骨基因表达有明显的促进作用,为HMGB1应用于临床提供实验依据。

Objective：To explore the efl＇ect of High mobility group box 1 （HMGBI） to bone marrow stromal cells （BM- SCs） from rats on prolit^ration,osteogenic/osteoclastic differentiation, nethod：BMSCs were obtained from wistar rats. After 14 days osteogenic inducing cuhure,sufficient cells were expanded for the following experiments. MTF analysis was condueted to BMSCs in osteogenic inducing medium containing 0,100,500 ng/mL HMGB1. Osteogenic differentiation of BMSCs cuhured in osteogenic medium supplemented with or without HMGB1 were measured by alkaline phosphatase（ALP） staining and RT- PCR analysis on genes including osteocalcin（OCN）,osteoprotegerin（OPG） and receptor activator of NF-kB ligand（RANKL） on day 7. Result：BMSCs treated with HMGB1 （100 and 500 ng/mL） proliferated faster than the cells cultured in osteogenic medium on day 3 and 5. And the genes expression including RANKL and RANKL/OPG was significantly enhanced. Howev- er, ALP staining and genes expression including OCN and OPG were not significantly enhanced by HMGB1 in the concentra- tion of 100 and 500 ng/mL. Conclusion： The use of proper concentrations of HMGB1 enhanced the proliferation and os- teoclastic differentiation abilities of BMSCs, which provided experimental basis for the clinical application of HMGB1.