Ca^(2+)信号对1α,25-(OH)_2D_3调控破骨细胞形成及骨吸收活性的影响  

EFFECTS OF CA^(2+) SIGNAL ON OSTEOCLASTS FORMATION AND BONE RESORPTION INDUCED BY 1α, 25-(OH)_2D_3

作　　者：仝锡帅[1,2,3] 顾建红[1,3] 王东[1,3] 陈阳[1,3] 卞建春 刘宗平[1,3] 

机构地区：[1]扬州大学兽医学院,扬州225009 [2]江苏省高邮市高邮镇农业服务中心,扬州225601 [3]江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009

出　　处：《营养学报》2015年第5期470-474,共5页Acta Nutrimenta Sinica

基　　金：国家自然科学基金资助项目(No.31172373;31302154);江苏高校优势学科建设工程资助项目;江苏省高校自然科学基金项目(No.13KJB230002);教育部博士点基金新教师类项目(No.20133250120002)

摘　　要：目的探讨Ca2+信号对1α,25-二羟维生素D3(1α,25-(OH)2D3)调控破骨细胞(osteoclast,OC)形成及骨吸收活性的影响。方法在巨噬细胞集落刺激因子(macrophage colony-stimulating factor,M-CSF)及核因子-κB受体活化因子配体(receptor activator of nuclear factor-κB ligand,RANKL)联合诱导RAW264.7细胞分化为OC的基础上,添加10-8 mol/L 1α,25-(OH)2D3,并设Ca2+螯合剂1,2-双(2-氨基苯氧基)乙烷-N,N,N`,N`-四乙酸四乙酸甲酯(BAPTA-AM)干预组,通过抗酒石酸酸性磷酸酶(TRAP)染色鉴定OC的形成,环境扫描电镜观察牛皮质骨片吸收陷窝。结果添加10-8 mol/L 1α,25-(OH)2D3可显著增加OC数量,促进骨吸收活性。然而,与添加10-8 mol/L 1α,25-(OH)2D3比较,5μmol/L BAPTA-AM干预可显著降低OC数量,抑制骨吸收活性。结论 10-8 mol/L 1α,25-(OH)2D3能够促进OC形成和骨吸收活性,此过程涉及Ca2+信号机制。Objective To study the effects of Ca2＋ signal on osteoclasts （OC） formation and bone resorption induced by 1α, 25-dihydroxyvitamin D3 [1α, 25-（OH）2D3] in vitro. Methods RAW264.7 cells were induced to form osteoclasts by 25μg/L macrophage colony-stimulating factor （M-CSF） and 30 p.g/L receptor activator of nuclear factor-kB ligand （RANKL） in this study. The RAW264.7 cells were divided into 6 groups： A group without any treatment, B group treated with M-CSF and RANKL, C group treated with M-CSF, RANKL and 10-8 mol/L 1α, 25-（OH）2D3, D group treated with M-CSF, RANKL, 10.8 mol/L 1α, 25-（OH）2D3 and 5gmol/L 1,2-Bis（2-aminophenoxy）-ethane-N,N,N＇N＇-tetraacetic acid tetrakis （BAPTA-AM）, E group treated with M-CSF, RANKL and 5gmol/L BAPTA-AM, F group treated with M-CSF, RANKL and 0.01% ethanol. Then, the formation and differentiation of osteoclasts were confirmed via tartrate resistant acid phosphatase （TRAP） staining and bone absorption analysis was performed by environmental scanning electron microscope. Results 10.8 mol/L 1α, 25-（OH）2D3 increased the number of OC and improved bone resorption of TRAP-positive osteoclasts significantly. However, compared to the group only treated with 1α, 25-（OH）2D3, the group with 1α, 25-（OH）2D3 plus BAPTA-AM showed reduced formation of OC and bone resorption of TRAP-positive osteoclasts significantly. Conclusion 10-8 mol/L let, 25-（OH）2D3 can enhance the formation and bone resorption of osteocalsts, and Ca2＋ signal transduction mechanism was involved in this process.

关 键 词：Ca2+信号 1α 25-(OH)2D3 破骨细胞 骨吸收 RAW264.7细胞 

分 类 号：Q25[生物学—细胞生物学]