碳酸酐酶1对骨形成作用的研究  

Carbonic Anhydrase 1 Is Involved in the Process of Bone Formation

作　　者：郑亚冰[1] 王林[1] 常建芳[2] 王月俭[1] 韩金祥[1] 常晓天[1] 

机构地区：[1]山东省医学科学院山东省医药生物技术研究中心,济南250062 [2]济南军区总医院生殖医学中心,济南250031

出　　处：《中国生物工程杂志》2012年第4期22-27,共6页China Biotechnology

基　　金：国家自然科学基金(30671949;30972720);国家基础研究发展计划(2010CB529105);山东泰山学者;山东省科学和科技(2009ZHZX1A1004)资助项目

摘　　要：目的:为了研究碳酸酐酶1(carbonic anhydrase I,CA1)在生物矿化和骨形成过程中的作用。方法:选择HeLa细胞为研究对象,用地塞米松(dexamethasone,DEX)、β-磷酸甘油(β-glycerophosphate,β-GP)和维他命C(ascorbic acid,AA)诱导其钙化,茜素红染色法观察矿化结节的形成,荧光定量PCR检测细胞骨化标志蛋白Runx2/cbfa1的表达。CA1表达水平的变化采用Western blot和荧光定量PCR的方法检测。同时用碳酸酐酶抑制剂乙酰唑胺处理HeLa细胞,观察CA1表达被抑制后对HeLa细胞钙化的影响。结果:HeLa细胞在诱导后形成大量矿化结节,Runx2/cbfa1转录水平明显升高,显示DEX、β-GP和AA可诱导HeLa细胞钙化和骨化,HeLa细胞钙化后CA1表达明显增高。HeLa细胞经乙酰唑胺处理后,CA1的表达量减少,矿化结节形成明显减少,说明抑制CA1表达可以抑制HeLa细胞矿化和骨化过程。结论:CA1在生物矿化和新骨形成过程中起着重要作用。Objective： Carbonic anhydrase I（CA1） not only enhances the hydration reaction but also promotes the formation of CaCO3,which is an essential step for new bone formation.However,there is no direct data to demonstrate the involvement of CA1 in bio-mineralization in cells and tissues.The aim is to determine the role of CA1 involvement in bio-mineralization and ossification with cultured cells.Methods： HeLa cells were incubated with dexamethasone（DEX）,β-glycerophosphate（β-GP） and ascorbic acid（AA） that can induce bio-mineralization under physiological condition.The cellular calcification was determined by Alizarin Red-S staining.The expression of Runx2/cbfa1,a protein marker of ossification,was detected in the process of bone formation by real-time PCR.The CA1 expression was determined by Western blot.Acetazolamide,an anti-carbonic anhydrase drug was also added to the culture to determine the role of CA1 in bio-mineralization and new bone formation.Results： Following the stimulation,HeLa cells produced a great amount of calcium-rich deposits.Increased transcription of Runx2/cbfa1 was also detected in the HeLa cells,indicating that the DEX,β-GP and AA mixture launched the process of bio-mineralization and bone formation in the cells.CA1 has a significantly increased expression during this process.Following treatment of acetazolamide,the expression of CA1 considerably declined,and calcium nodule formation was significantly decreased.Conclusion： The above findings suggested that CA1 participates in bio-mineralization and ossification,and plays an important roles in the process.

关 键 词：生物矿化 碳酸酐酶1 成骨诱导 

分 类 号：Q819[生物学—生物工程]