CAJ | 학술논문

目的观察腺相关病毒(AAV)介导的成骨蛋白-1(OP-1)和性别决定区Y框蛋白9(SOX9)双基因体内转染对兔退变椎间盘的影响.方法针刺损伤法制作兔退变椎间盘模型,于4周后,分别于造模椎间盘髓核内注射20μl双基因混合液(1∶1)、重组AAV(rAAV)-OP-1、rAAV-SOX9、rAAV-增强型绿色荧光蛋白(EGFP)、磷酸盐缓冲液(PBS),于转染后9周行MRI观察椎间盘变化,逆转录-聚合酶链反应(RT-PCR)检测Ⅱ型胶原mRNA及蛋白多糖mRNA的变化.结果 MRI显示双基因组T2加权像信号强度明显恢复,与SOX9、OP-1组比差异有统计学意义(P＜0.05)；RT-PCR证实双基因组Ⅱ型胶原及蛋白多糖mRNA表达量均明显高于单一SOX9及OP-1组(P＜0.05).结论 SOX9及OP-1双基因联合转染治疗退变椎间盘组织具有明显作用；双基因治疗椎间盘退变具有协同效用.
목적 관찰선상관병독(AAV)개도적성골단백-1(OP-1)화성별결정구Y광단백9(SOX9)쌍기인체내전염대토퇴변추간반적영향.방법 침자손상법제작토퇴변추간반모형,우4주후,분별우조모추간반수핵내주사20μl쌍기인혼합액(1∶1)、중조AAV(rAAV)-OP-1、rAAV-SOX9、rAAV-증강형록색형광단백(EGFP)、린산염완충액(PBS),우전염후9주행MRI관찰추간반변화,역전록-취합매련반응(RT-PCR)검측Ⅱ형효원mRNA급단백다당mRNA적변화.결과 MRI현시쌍기인조T2가권상신호강도명현회복,여SOX9、OP-1조비차이유통계학의의(P＜0.05)；RT-PCR증실쌍기인조Ⅱ형효원급단백다당mRNA표체량균명현고우단일SOX9급OP-1조(P＜0.05).결론 SOX9급OP-1쌍기인연합전염치료퇴변추간반조직구유명현작용；쌍기인치료추간반퇴변구유협동효용.
Objective To investigate the biological effect of the recombinant adeno-associated virus (rAAV)-mediated osteogenic protein-1 (OP-1) gene and sex determining region Y-box 9 (SOX9) gene transfer in rabbit degenerative intervertebral discs on MRI,collagen type Ⅱ and proteoglycan levels.Methods The lumbar disc degeneration model was established in the rabbits using percutaneous needle puncture.Twenty-five mature New Zealand white rabbits (13 males and 12 females,weighing 1.7-2.8 kg)were randomly divided into 5 groups:group A receiving 20 μl rAAV-OP-1 and rAAV-SOX9 (1 × 105 PFU); group B receiving 20 μl rAAV-OP-1; group C receiving 20 μl rAAV-SOX9; group D receiving 20 μl rAAV-enhanced green fluorescent protein(EGFP) ; and group E receiving 20 μl PBS.Nine weeks later,the changes of the intervertebral generative discs were observed by MRI,and the changes in type Ⅱ collagen mRNA and proteoglycan mRNA were examined by using reverse transcriptase-polymerase chain reaction (RT-PCR).Results The intervertebral generative discs recovered in a great measure on MRI,more significantly in double gene group than in single gene group (P ＜ 0.05).The nucleus pulposus tissues from double gene group exhibited an increased expression of proteoglycan mRNA and Col2al mRNA as compared with single gene group (P ＜ 0.05).Conclusion The rAAV-mediated OP-1 and SOX9 transfer is effective in the treatment of rabbit degenerative intervertebral discs with synergetic effects.