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首页> 外文期刊>International journal of molecular medicine >Treatment of rabbit intervertebral disc degeneration with co-transfection by adeno-associated virus-mediated SOX9 and osteogenic protein-1 double genes in vivo
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Treatment of rabbit intervertebral disc degeneration with co-transfection by adeno-associated virus-mediated SOX9 and osteogenic protein-1 double genes in vivo

机译:腺相关病毒介导的SOX9和成骨蛋白-1双基因联合转染体内治疗兔椎间盘退变

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摘要

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.
机译:腰椎间盘退变是腰痛和腿痛的常见原因,它会影响患者的身心健康并增加社会负担。进行这项研究以观察腺相关病毒(AAV)介导的成骨蛋白1(OP1)和SOX9双基因共转染在体内兔椎间盘退变中的生物学作用。将动物随机分组为椎间盘退变模型。将20μl双基因混合溶液,OP1,SOX9,增强型绿色荧光蛋白(EGFP)和PBS缓冲液注入各组椎间盘后,进行X射线分析,磁共振成像(MRI),逆转录PCR(RT-在手术的第3、6和9周进行PCR和Western blotting。在转染的第3、6和9周,X射线和MRI显示A,B和C组的椎体高度和T2加权信号强度显着恢复,而椎间隙和T2加权信号显着差异A组与B,C组之间的差异有统计学意义(P <0.05)。 RT-PCR和western blotting结果显示,A,B,C组Ⅱ型胶原蛋白和蛋白聚糖mRNA表达上调。A组明显高于其他各组(P <0.05)。重组AAV介导的SOX9和OP1双基因转染显着改善了变性椎间盘的高度,并显着促进了变性椎间盘蛋白聚糖和II型胶原的高表达。因此可以得出结论,双基因疗法具有协同作用。

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