白介素-6与1,25(OH)_2维生素D_3联合作用下大鼠骨髓破骨细胞的体外诱导培养

目的观察联合应用白介素(IL)-6和1,25(OH)2D3对大鼠骨髓破骨细胞生成诱导的影响。方法采用4周龄SD大鼠无菌条件下取出股骨,剪去两骺端,以α-MEM培养液将骨髓细胞冲出,然后将细胞悬液接种于预置盖玻片或牙本质片的24孔培养板内,24 h后换液。试验分为4组,A组:不加任何诱导因子;B组:加入IL-6(10 U/ml);C组:加入1,25(OH)2D3(1×10-8mol/L);D组:加入IL-6(10 U/ml)和1,25(OH)2D3(1×10-8mol/L)。每组各6孔,于培养的第7天进行多核细胞计数。结果培养1周左右IL-6与1,25(OH)2D3在体外均可单独诱导骨髓破骨细胞的形成,但D组多核细胞数与B、C组相比差异有显著性(P<0.05)。结论IL-6与1,25(OH)2D3联合作用下对骨髓破骨细胞生成的诱导效果明显高于单因素诱导效果。

IL-6 and 1,25(OH)_2D_3 combined induces formation of osteoclast in rat bone marrow culture

Objective To investigate the effects of the combination of IL-6 and 1,25(OH)_2D_3 on the osteoclast formation in rat bone marrow culture.Methods Bone marrow cells were isolated from 4-week-old SD rats and suspended in a-MEM,then added to the wells of 24-well plates with cover-slips or dentin slices.After 24 hours,the media were replaced with IL-6 and/or 1,25(OH)_2D_3.TRAP(tartrate-resistant acid phosphatase) staining was used to indentify osteoclast.The bone resorption pit on dentin slices were examined with inverted phase contrast microscope.Results When bone marrow cells were cultured for up to 7 days,either in the presence of IL-6(10 U/ml)or 1,25(OH)_2D_3(1×10~(-8)mol/L),mononuclear cells were found to form TRAP-positive multinuclear cells.But in the presence of IL-6(10 U/ml)and 1,25(OH)_2D_3(1×10~(-8)mol/L),the number of TRAP-positive multinuclear cells increased.Conclusion The effects of the combination of IL-6 and 1,25(OH)_2D_3 on the osteoclast formation are more powerful than a single factor.