腺相关病毒载体介导结缔组织生长因子对体外转染恒河猴和人腰椎间盘细胞蛋白多糖和Ⅱ型胶原影响的比较  

Comparison of the biological effe cts of adeno-associated virus vectors of connective tissue growth factor on the proteoglycan level and type Ⅱ collagen between disc cells of rhesus monkey and human cultured in vitro

作　　者：刘勇[1] 孔杰[1] 魏见伟[1] 胡有谷[1] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,青岛266003

出　　处：《中国骨与关节外科》2009年第1期50-56,共7页Chinese Journal of Bone and Joint Surgery

基　　金：国家自然科学基金资助(30271318)

摘　　要：目的比较腺相关病毒载体介导结缔组织生长因子(connective tissue growth factor,CTGF)对体外培养的恒河猴和人腰椎间盘细胞转染后蛋白多糖含量和Ⅱ型胶原的影响。方法将恒河猴及成人腰椎间盘髓核细胞进行体外培养,应用rAAV2-CTGF体外转染细胞,分别通过^(35)S标记蛋白多糖的方法和Ⅱ型胶原的SP-ABC免疫组化法检测腺相关病毒载体介导生长因子对椎间盘细胞蛋白多糖合成和Ⅱ型胶原的影响。结果恒河猴椎间盘细胞能进行体外培养并传代,其形态学特性与培养的成人椎间盘细胞相似,rAAV2-CTGF与对照组相比可促进恒河猴和人椎间盘髓核细胞的蛋白多糖生物合成(P<0.01),恒河猴和成人相比生长因子对于蛋白多糖的促进作用无明显差异(P>0.05)。结论成功培养恒河猴及成人椎间盘细胞,腺相关病毒载体介导CTGF能显著促进髓核细胞蛋白多糖的合成,生长因子对恒河猴和人椎间盘细胞蛋白多糖含量的影响具有很大的相似性。Objective To investigate the different effects of adeno-associated vires vectors of connective tissue growth factor （CTGF） on the content of proteoglycan and type Ⅱ collagen of disc cells of monkey and human. Methods Both monkey and human nucleus pulpouses were isolated and cultured. The adeno-associated virus vectors of CTGF were transfected into the disc cells respectively. The biological effects of CTGF were detected by 35S detection and immunohistochemiscal method. Results The disc cells of rhesus monkey and human were cultured and subcultured. The morphological characteristicsof cultured disc cells of monkey were similar to those of human. Compared with control group, the different groups which transfected with CTGF showed significantly increased levels of proteoglyean content and type Ⅱ collagen （P〈0.05）. There was no significant difference between rhesus monkey and human on the contents of proteopglycan and type Ⅱ collagen （ P 〉 0. 05 ）. Conciusion Disc cells of rhesus monkey as well as human were cultured successfully. The adeno-associated virus vectors of CTGF-TIMP1 can remarkably increase the level of proteoglycan and type Ⅱ collagen of disc cells. The effects of growth factor on proteoglycan incorporation and content of type Ⅱ collagen of disc cells of rhesus monkey are similar to those of human.

关 键 词：结缔组织 生长因子 椎间盘 胶原Ⅱ型 

分 类 号：R681.5[医药卫生—骨科学]