梁静;邓廉夫;朱雅萍;周琦;魏立;

• 1:上海交通大学瑞金医院上海市伤骨科研究所

摘要(Abstract)：

[目的]观察检测低氧和常氧条件下原代和传代后软骨细胞各特异性基因及蛋白表达的改变。[方法]采用3～5日龄C57BL/6小鼠,取四肢关节软骨,经机械分离和酶消化法获得关节软骨细胞,将原代细胞P0和传代后细胞P1、P2分别在普通和低氧培养箱中培养2d,用RT-PCR检测II型胶原、可聚蛋白聚糖和sox9特异性基因及Ihh、PTHrP、bmp4、wnt5a分化相关基因的表达差异,原代软骨细胞用免疫组化和阿利新蓝染色检测II型胶原、可聚蛋白聚糖的蛋白表达。[结果]低氧条件下,免疫组化显示II型胶原和可聚蛋白聚糖的表达较普通培养增强,II型胶原、wnt5a的基因表达增强,Ihh的表达降低;传代后软骨细胞的蛋白、特异性基因和分化相关基因表达未见明显差异。[结论]低氧条件下,原代软骨细胞的II型胶原表达增强,且可能主要是受Ihh、wnt5a等分化相关基因的调控。短期单层培养方式不能明显改善、恢复特异性基因表达降低的传代后软骨细胞表型。

关键词(KeyWords)： 软骨细胞;低氧;Ⅱ型胶原;蛋白聚糖

Abstract:

Keywords:

基金项目(Foundation): 上海交通大学医学院科技基金项目(编号:05XJ21035)

作者(Author): 梁静;邓廉夫;朱雅萍;周琦;魏立;

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DOI:

参考文献(References)：

• [1]Lee RB,Urban JP.Evidence for a negative pasteur effect in articular cartiage[J].Biochem J,1997,1:95-102.
• [2]Hauselmann HJ,Fernandes RJ,Mok SS,et al.Phenotypic stability of bovine articular chondrocytes after long-term culture in alginate beads[J].J Cell Sci,1994,1:17-27.
• [3]Hirao M,Tamai N,Tsumaki N,et al.Oxygen tension regulates chon-drocyte differentiation and function during endochondral ossification[J].J Biol Chem,2006,41:31079-31092.
• [4]Murphyl CL,Polak JM.Control of human articular chondrocyte differ-entiation by reduced oxygen tention[J].J Cell Physiol,2004,3:451-459.
• [5]Mackie EJ,Ahmed YA,Tatarczuch L,et al.Endochondral ossification:how cartilage is converted into bone in the developing skeleton[J].Int J Biochem Cell Biol,2008,1:46-62.
• [6]ShumL,Wang X,Kane AA,et al.BMP4promotes chondrocyte prolif-eration and hypertrophy in the endochondral cranial base[J].Int J Dev Biol,2003,6:423-431.
• [7]Yingzi Y,Lilia T,Heuijung L,et al.Wnt5a and Wnt5b exhibit dis-tinct activities in coordinating chondrocyte proliferation and differentia-tion[J].Development,2003,5:1003-1015.