TNF-α诱导后的小鼠成骨细胞对破骨前体细胞增殖的影响

摘要

目的：通过将小鼠成骨细胞与破骨前体细胞间接共培养,研究经肿瘤坏死因子-α( tumour necrosis fac-tor-α,TNF-α)诱导后模拟炎症状态下的成骨细胞对破骨前体细胞增殖功能的影响。方法按照TNF-α10 ng/mL的浓度对鼠成骨细胞分别培养2、4、8、16 h,以诱导其进入炎症状态,并分别提取其上清液,与加入了核因子κB受体活化因子配体(receptor activator for nuclear factor-κ B ligand,RANKL)30μg/L 的鼠单核细胞共培养,利用激光共聚焦显微镜观察和细胞计数试剂盒(cell counting kit-8,CCK-8)法进行增殖活性检测。结果 TNF-α诱导2 h后的成骨细胞上清液与加入RANKL 的单核细胞共培养时,细胞增殖活性实验组与对照组之间差异无统计学意义(P>0.05),诱导4 h实验组破骨前体细胞的增殖活性高于对照组,8、16 h实验组破骨前体细胞的增殖活性低于对照组。结论短暂炎症刺激下,成骨细胞可能对破骨前体细胞增殖有促进作用；长时间炎症刺激下,成骨细胞对破骨前体细胞的增殖可能有抑制作用。%Objective To study the influence of the TNF-α induced mouse osteoblast under the inflammation condi-tion on the function of the proliferation of osteoclast precursor cells by the indirect co-culture of the mouse osteoblast and osteoclast precursor cells. Methods The mouse osteoblast were cultured for 2, 4, 8, 16 h respectively according to the TNF-α10 ng/mL concentration in order to be induced into inflammatory state. Its supernatant was extracted respectively and co-cultured with mouse monocyte with the addition of 30 μg/L activator for nuclear factor- receptor nuclear factor kappa ligand ( RANKL) . The proliferation activity was detected by laser scanning confocal microscopy and cell counting kit-8 cell ( CCK-8) method. Results When the supernatant induced by TNF-α for 2 h was co-cultured with monocyte with RANKL, there was no significant difference between the experimental group and the control group. The proliferation of primary osteoclast in experimental group was significant higher than control group when the osteoblast was induced at 4 h. However, the proliferation of primary osteoclast in experimental group was significant lower than control group when the osteoblast was induced at 8, 16 h. Conclusion The osteoblasts may promote the proliferation of osteoclast precursor cells under transient inflammation stimulation. However, the osteoblast may have inhibitory effects on the proliferation of osteoclast precursor cells under the prolonged inflammatory stimulation.