FGF2对破骨细胞体外分化以及骨吸收功能的直接调控作用  

Fibroblast growth factor 2 directly regulates osteoclast differentiation and bone resorption function

作　　者：李晓刚[1] 苏楠[1] 唐玉彬[1] 温轩[1] 唐浚洲 陈思宇[1] 李灿[1] 杜晓兰[1] 陈林[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所创伤实验室,骨代谢与修复中心,创伤,烧伤与复合伤国家重点实验室,重庆400042

出　　处：《第三军医大学学报》2014年第7期636-639,共4页Journal of Third Military Medical University

基　　金：国家重点基础研究发展计划(973计划;2014CB942904);国家自然科学基金(81270012;81170809);国家重点实验室自主研究课题(SKLZZ201017)~~

摘　　要：目的研究成纤维细胞生长因子2(fibroblast growth factor 2,FGF2)对于破骨细胞(osteoclast,OC)体外分化与功能的直接调控作用。方法分离野生型C3H/HeJ小鼠骨髓单核细胞,利用核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)以及巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)诱导向破骨细胞分化,同时在实验组中加入FGF2,观察骨髓单核细胞向破骨细胞分化发育过程,鬼笔环肽(phalloidine)染色观察破骨细胞肌动蛋白环(actin ring)变化,Real-time PCR检测破骨细胞Trap、Mmp9、Ctsk以及Clcn7表达,破骨细胞与骨片共培养检测骨吸收功能变化,并用Western blot检测相关信号通路分子表达情况。结果①TRAP染色显示实验组与对照组破骨细胞数量无明显差异(P>0.05);②鬼笔环肽染色提示实验组与对照组肌动蛋白环均正常;③实验组Trap、Mmp9及Ctsk mRNA表达量均高于对照组(P<0.05),而Clcn7 mRNA表达2组无明显差异;④体外骨片吸收实验提示实验组骨吸收陷窝面积显著大于对照组(P<0.05);⑤Western blot检测实验组磷酸化细胞外信号调节激酶(ERK)较对照组表达明显增多。结论 FGF2不影响破骨细胞前体细胞向破骨细胞的分化以及肌动蛋白环的形成,但可以通过ERK信号通路正向调控成熟破骨细胞的骨吸收功能。Objective To determine the effect of fibroblast growth factor 2 （FGF2） on the differentia tion of bone marrow monocytes （BMMCs） to osteoclast and bone resorption function. Methods After BMMCs were isolated from the bone marrow of femurs and tibiae from C3H mice, the cells were treated with receptor activator of nuclear factor-kappa B ligand （RANKL） and macrophage colony stimulating factor （M-CSF）. Experimental and control groups were separated by addition of FGF2 or not. The differentiation and maturation of osteoclast precursors were observed by TRAP staining, and the osteoclast actin ring of both group were analyzed by phallodine staining. The expression of Trap, Mmp9, Ctsk and Clcn7 at mRNA level were detected by real-time PCR. The function of bone resorption was measured by bovine bone slice and osteoclast coculture. The effect of FGF2 on activity of ERK protein in the osteoclasts was analyzed by Western blotting. Results There was no significant difference in differentiation and maturation of osteoclast precursors between 2 groups influenced. Compared with control group, the bone resorption lacuna area of FGF2 treated group was larger （P 〈 0.05）. Finally, Western blot analysis showed that treatment with FGF2 induced enhanced activity of phosphorylated ERK in cultured osteoclasts. Conclusion FGF2 cannot influence the differentiation of osteoclast precursors or formation of actin ring, but can upreaulate the function of ostenclastic resorption via anERK-related pathway.

关 键 词：破骨细胞 成纤维细胞生长因子2 细胞外信号调节激酶 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]