Department of Spine Surgery, Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong 266003, P.R. China.
Int J Mol Med. 2013 Nov;32(5):1063-8. doi: 10.3892/ijmm.2013.1497. Epub 2013 Sep 17.
Degeneration of the lumbar intervertebral disc is a common cause of low back pain and leg pain that affects the physical and mental health of the patient and increases the social burden. This study was performed to observe the biological effects of adeno-associated virus (AAV)-mediated osteogenic protein-1 (OP1) and SOX9 double gene co-transfection in rabbit intervertebral disc degeneration in vivo. The animals were randomly grouped into models of disc degeneration. After injecting 20 µl of double-gene mixed solution, OP1, SOX9, enhanced green fluorescent protein (EGFP) and PBS buffer into the disc of each group, X-ray analysis, magnetic resonance imaging (MRI), reverse transcription PCR (RT-PCR) and western blotting were performed on the 3rd, 6th and 9th week of surgery. On the 3rd, 6th and 9th week of the transfection, X-ray and MRI showed that the intervertebral height and T2-weighted signal intensity were restored significantly in groups A, B and C, whereas significant differences in intervertebral space and T2-weighted signal intensity were observed between group A and groups B and C (P<0.05). RT-PCR and western blotting showed that the expression of type II collagen and proteoglycan mRNA was upregulated in groups A, B and C. The expression in group A was significantly higher than that in the other groups (P<0.05). Recombinant AAV-mediated SOX9 and OP1 double-gene transfection significantly ameliorated the height of the degenerative intervertebral disc and significantly promoted the high expression of degenerative disc proteoglycan and type II collagen. It can therefore be concluded that dual-gene therapy has a synergistic effect.
腰椎间盘退变是引起腰痛和腿痛的常见原因,影响患者的身心健康,增加社会负担。本研究旨在观察腺相关病毒(AAV)介导的骨形成蛋白-1(OP1)和 SOX9 双基因共转染对兔椎间盘退变的生物学效应。将动物随机分为椎间盘退变模型组。每组椎间盘内注射 20 μl 双基因混合液、OP1、SOX9、增强型绿色荧光蛋白(EGFP)和 PBS 缓冲液后,分别于术后第 3、6、9 周进行 X 线分析、磁共振成像(MRI)、反转录 PCR(RT-PCR)和 Western blot 检测。转染后第 3、6、9 周,X 线和 MRI 显示 A、B、C 组的椎间盘高度和 T2 加权信号强度显著恢复,而 A 组与 B、C 组的椎间间隙和 T2 加权信号强度差异有统计学意义(P<0.05)。RT-PCR 和 Western blot 显示 A、B、C 组 II 型胶原和蛋白聚糖 mRNA 表达上调,A 组表达明显高于其他组(P<0.05)。重组 AAV 介导的 SOX9 和 OP1 双基因转染显著改善退变椎间盘的高度,显著促进退变椎间盘蛋白聚糖和 II 型胶原的高表达,表明双基因治疗具有协同作用。