降钙素对体外培养破骨细胞功能的影响  

In vitro effects of calcitonin on osteoclasts

作　　者：张秀珍[1] 韩峻峰[1] 钱国峰[1] 

机构地区：[1]同济大学附属同济医院内分泌科,上海200065

出　　处：《中华内分泌代谢杂志》2004年第2期158-160,共3页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金资助项目 (30 0 70 357)

摘　　要：目的　观察降钙素对体外培养破骨细胞功能以及成骨细胞的骨保护素 (OPG)、细胞核因子 κB受体活化因子配基 (RANKL)的表达的影响。方法　分别用酶消化法和机械分离获得新生SD大鼠成骨细胞、破骨细胞 ,在培养液中分别加入不同浓度的降钙素 ,以抗酒石酸酸性磷酸酶 (TRAP)染色观察破骨细胞数目、形态 ,Image ProPlus图像软件分析骨片上骨吸收陷窝的数目和面积。用RT PCR方法观察成骨细胞OPG、RANKL的表达。结果　各组破骨细胞数目随着降钙素浓度增加而减少 (P <0 .0 5 )。骨片培养5天 ,吸收陷窝计数的结果显示 ,10 -14 mol/L以上浓度降钙素对破骨细胞吸收功能均有明显抑制作用 ,并呈剂量相关性。 10 -12 mol/L组陷窝面积为对照组的 49% (P <0 .0 1) ,10 -8mol/L组为对照组的 3 0 % (P <0 .0 1)。降钙素组破骨细胞OPGmRNA表达较对照组升高 ,RANKL降低 (均P <0 .0 5 )。结论　除了直接作用于破骨细胞外 ,降钙素还通过影响成骨细胞上OPG、RANKL的分泌间接调控破骨细胞功能 ,抑制骨吸收。Objective To observe the effects of calcitonin on osteoclast activity and expressions of osteoprotegrin (OPG) and receptor activator of NF-κB ligand (RANKL) in vitro. Methods Osteoclasts and their precursors were mechanically isolated from long bones of SD rats and incubated on bone sheet in MEM with different concentration of calcitonin. Number, morphology of the osteoclasts were detected with tartrate-resistance acid phosphatase (TRAP) staining. Image-Pro Plus counted the resorption pits and areas on slices, stained by toludine blue. RT-PCR was used to examine the expressions of OPG and RANKL of osteoclast. Results Number of osteoclasts were decreased with increasing CT concentrations. After incubation for 5 days, resorption pits were decreased in a concentration-dependent manner (P<0.05). Areas of the resorption pits at 10 -12mol/L and 10 -8mol/L were 49% and 30% of those of control group respectively. OPG mRNA was increased, but RANKL mRNA was down-regulated in osteoclasts by calcitonin (P<0.05). Conclusion Except for inhibiting osteoclast resorption activity in vitro directly, calcitonin could also affected osteoclast activity by regulating expressions of OPG and RANKL.

关 键 词：降钙素 体外培养 破骨细胞 成骨细胞 细胞培养 

分 类 号：R96[医药卫生—药理学]