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腺相关病毒介导的骨形态发生蛋白-7和SOX9对人退变椎间盘细胞的体外共转染

Adeno-associated virus-mediated BMP-7 and SOX9 in vitro co-transfection of human degenerative intervertebral disc cells.

作者信息

Ren X F, Diao Z Z, Xi Y M, Qi Z H, Ren S, Liu Y J, Yang D L, Zhang X, Yuan S L

机构信息

Department of Orthopaedics, Affiliated Hospital of Qingdao University, Qingdao, China.

Department of Orthopaedics, Affiliated Hospital of Qingdao University, Qingdao, China

出版信息

Genet Mol Res. 2015 Apr 22;14(2):3736-44. doi: 10.4238/2015.April.22.1.

Abstract

Bone morphogenetic protein-7 (BMP-7) and SOX9 are important transcription factors in chondrogenesis. In this study, we examined the biological function of the adeno-associated virus (AAV)-mediated BMP-7 and SOX9 double gene in vitro co-transfection of nucleus pulposus cells of human degenerative intervertebral disc. Human intervertebral disc nucleus pulposus cells were cultured in vitro and subcultured for 5 generations. Using rAAV-BMP-7 and rAAV-SOX9 AAV2-type AAV viruses, the cells were divided into 4 groups: blank normal, BMP-7 transfection, SOX9 transfection, and BMP-7 and SOX9 co-transfection. After 48 h, expression of type II collagen and its mRNA in nucleus pulposus cells was determined. The expression of type II collagen in BMP-7 transfection, SOX9 transfection, and co-transfection groups was up-regulated to varying degrees compared to the blank control group. The type II collagen mRNA level expression in the co-transfection group was significantly higher than in other groups (P < 0.05). AAV-mediated BMP-7 and SOX9 in vitro co-transfection can promote the synthesis of type II collagen in nucleus pulposus cells in the human degenerative intervertebral disc. Double-gene therapy has a synergistic effect in the treatment of intervertebral disc degeneration.

摘要

骨形态发生蛋白-7(BMP-7)和SOX9是软骨形成过程中的重要转录因子。在本研究中,我们检测了腺相关病毒(AAV)介导的BMP-7和SOX9双基因在人退变椎间盘髓核细胞体外共转染中的生物学功能。将人椎间盘髓核细胞进行体外培养并传代5次。使用rAAV-BMP-7和rAAV-SOX9 AAV2型腺相关病毒,将细胞分为4组:空白正常组、BMP-7转染组、SOX9转染组以及BMP-7和SOX9共转染组。48小时后,检测髓核细胞中Ⅱ型胶原蛋白及其mRNA的表达。与空白对照组相比,BMP-7转染组、SOX9转染组和共转染组中Ⅱ型胶原蛋白的表达均有不同程度上调。共转染组中Ⅱ型胶原蛋白mRNA水平表达显著高于其他组(P<0.05)。AAV介导的BMP-7和SOX9体外共转染可促进人退变椎间盘髓核细胞中Ⅱ型胶原蛋白的合成。双基因治疗在椎间盘退变的治疗中具有协同作用。

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