塞来昔布抑制骨髓间充质干细胞的增殖和成骨诱导分化

The effects of celecoxib on proliferation of human bone mesenchymal stem cells and differentiation into osteoblasts

目的探讨非甾体类抗炎药塞来昔布对人骨髓间充质干细胞（hBMSCs）增殖和分化成骨的影响方法骨髓取自1名健康男性，采用Ficoll—Paque密度梯度离心分离hBMSCs，并进行体外扩增。在细胞增殖和诱导过程中加入不同浓度（12．5、25．0、50．0、100．0μmol／L）的塞来昔布，采用细胞计数试剂盒（CCK8）方法测定塞来昔布对hBMSCs增殖的影响。使用地塞米松、维生素C等试剂诱导hBMSCs分化成骨，在hBMSCs成骨过程中加入100μmol／L塞来昔布，通过碱性磷酸酶（ALP）活性分析、钙定硅分析等疗法测定塞来占布对hBMSCs成骨的影响，通过Westernblot方法检测塞来昔布埘hBMSCs分化成骨关键转录因子Runx2表达的影响。结果塞来昔布的作用呈现剂量依赖性。12．5～100．0μmol／L摩来昔布对hBMSCs细胞增殖均有抑制作用，该作用随着药物浓度的增加而增强。大剂量的塞来昔布抑制hBMSCs分化成骨中的碱性磷酸酶和钙沉积，并抑制转录因子Runx2的表达。结论塞来昔布抑制hBMSCs增殖和成骨分化，并与剂最成正相关。长时间使用，尤其是大剂量使用塞来昔布的患者应考虑到其对成骨分化的抑制作用。

Objective To investigate tile effects of ceiecuxib, a kind of nonsteroidal anti-inflammatory drugs, on the proliferation of human bone mesenehymal stem cells （hBMSCs） and their differentiatiml into usteoblasts. Methods The bone marrow was harvested from a healthy male vohmteer. hBMSCs were isolated by density gradient eentrifugation with Fieoll-Paque and expanded in vitro. Celecoxib was added at the concentrations ranging from 10 μmol/L to 100 μmnl/L in the process of cellular proliferaticm and induction. The effect of celecoxib on the proliferation of hBMSCs was evaluated by CCK8 assay, hBMSCs were induced to differentiate into osteoblasts by dexamethasone aseorbie acid anti [3-glyeerophosphate. hBMSCs were treated with 100 μmol/L of celeeoxib in the process of osteogenesis before alkaline phosphatase （ALP） activities and cahium deposition were analyzed to determine the effect of celeeoxib on the differentiation into osteoblasts. The expression of Runx2, a critical transcription factor of osteogenesis, was analyzed by Western blot. Results Celeeoxib inhibited the hBMSCs proliferation in a dose-dependent manner. Such an inhibiting effeet of eelecoxib was observed at concentrations of 12.5μmol/l, to 100 μmol/L, intensified with the increased concentration. The ALP activities and calcium deposition were reduced by a high dose of Celeeoxib and the expression level of Runx2 was also suppressed. Conclusions Celeeoxib can suppress the proliferation and nsteogenesis of hBMSCs in a dose-dependent manoer. Long term administration of eelecnxib, particularly high dose usage, shouhl arouse concerns from both the patients and medical staff about the effect of celeeoxib on tile ostengenesis of hBMSCs.