环孢素A对成骨细胞分化的影响  

Effect of cyclosporin A on the osteoblastic differentiation

作　　者：宋丽华[1] 肖洲生[1] 潘玮[1] 余艳辉[1] 周宏灏[1] 

机构地区：[1]中南大学湘雅医学院临床药理学研究所分子药理学研究室,长沙410078

出　　处：《中华器官移植杂志》2003年第4期236-238,共3页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金资助项目 ( 30 1710 85)

摘　　要：目的　探讨环孢素A(CsA)对原代培养小鼠骨髓间质干细胞 (BMSCs)增殖及向成骨细胞分化的影响。方法　采用原代培养小鼠BMSCs模型 ,在培养液中加入不同浓度的CsA以及单独或联合使用左旋精氨酸 (L arginine)后 ,测定 [3 H] 甲基胸腺嘧啶 ([3 H] TdR)掺入率与细胞数 ,反映细胞增殖情况 ;测定碱性磷酸酶活性与钙沉积量 ,反映细胞向成骨细胞分化状态 ;用一氧化氮 (NO)试剂盒测定细胞培养基中NO释放量。结果　CsA呈剂量依赖性 (10 -6、3× 10 -6、6× 10 -6、10 -5mol/L)减少 [3 H] TdR掺入率 ;抑制细胞增殖 ;降低细胞碱性磷酸酶活性与钙沉积量 ;并伴随NO释放量减少。如果同时给予L arginine(10 -4mol/L) ,可完全逆转CsA(10 -6mol/L)的以上作用。结论　CsA可能通过抑制L arginine/NO通路而阻碍BMSCs增殖及向成骨细胞分化。Objective To study the in vitro effect of cyclosporin A (CsA) on the proliferation and osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells (BMSCs). Methods By using primary culture rat BMSCs model, [ 3H]-thymidine incorporation and cell number were measured after addition of CsA alone at different concentration or in combination with L-arginine in the culture medium. The osteoblastic differentiation in BMSCs was assessed by measuring alkaline phosphatase activity and calcium deposition. The effect of CsA on nitric oxide (NO) production was determined by measuring the stable end product of NO, nitrite, in the culture medium using commercial NO kit. Results CsA (10 -6 , 3×10 -6 , 6×10 -6 , 10 -5 mol/L) could decrease [ 3H]-thymidine incorporation and inhibit cell proliferation, reduce alkaline phosphatase activity and calcium deposition, and lower the NO production in the conditioned media in a dose-dependent manner. Concurrent treatment with L-arginine (10 -4 ?mol/L) could reverse the CsA (10 -6 ?mol/L)-mediated decrease in NO production, restore the proliferation and differentiation potential of BMSCs. Conclusion CsA blocked the proliferation and osteoblastic differentiation of mouse BMSCs through the inhibition of L-arginine/NO pathway.

关 键 词：环孢素A 免疫抑制剂 不良反应 骨质疏松症 成骨细胞 骨髓间质干细胞 一氧化氮 

分 类 号：R96[医药卫生—药理学]