腺病毒介导的hNT-3基因转染对兔前交叉韧带重建术后本体感觉恢复的影响

Effects of Adenovirus-mediated hNT-3 Gene Transfection on Proprioception Recovery after ACL Reconstruction

目的:探讨腺病毒介导的人神经营养素3(h NT-3)基因转染对兔前交叉韧带损伤重建术后本体感觉恢复的影响。方法:42只日本大耳白兔随机分成A、B、C三组,单侧后肢膝关节制造前交叉韧带损伤后自体半腱肌肌腱移植的动物模型。A、B两组自体半腱肌肌腱移植物内分别用微量注射器多点注射腺病毒载体Ad h NT-3与腺病毒空载体Ad GFP,C组注射生理盐水作为对照。术后48 h,分别从A、B、C三组中取自体肌腱移植物做冰冻切片,荧光显微镜观察绿色荧光蛋白(GFP)表达情况。术后8周、16周、24周,分别从A、B、C三组中取标本做电生理学指标检测、免疫组织化学染色和HE染色,观测体感诱发电位(SEPs)、肌电图(EMG)和本体感受器。结果:荧光显微镜下观察,A、B两组中可见绿色荧光蛋白表达,C组中未见。相同的时间点,A组本体感受器的数目多于B、C两组,体感诱发电位和肌电图的潜伏期短于B、C两组,波幅高于B、C两组,差异有统计学意义(P<0.05),而B、C两组中各观测指标相比较无明显统计学差异(P>0.05)。结论:腺病毒介导的h NT-3基因转染能够促进前交叉韧带损伤重建术后自体肌腱移植物内本体感受器的再生和本体感觉的恢复。

Objective To investigate the effect of adenovirus-mediated human neurotrophin 3（Adh NT-3） gene transfection on the proprioception recovery after the reconstruction of the anterior cruci-ate ligament（ACL） in rabbits. Methods Forty-two Japanese white rabbits were introduced the anteriorcruciate ligament reconstruction using the autologous semitendinosus tendon, and randomly divided intogroup A, B and C. Group A and B were respectively injected Ad h NT-3 and adenovirus-mediatedgreen fluorescent protein（Ad GFP）, while group C were injected saline as the control. Forty-eighthours after the operation, frozen sections were made from autologous tendon grafts to observe the ex-pression of green fluorescent protein（GFP） using the fluorescence microscopy. At 8, 16 and 24 weeksafter the surgery, specimens were measured somatosensory evoked potential（SEPs） and electromyography（EMG） using the electrophysiological method, as well as proprioceptors using the immunohistochemicaland haematoxylin-eosin（HE） staining. Results The GFP expression was found in group A and B underthe fluorescence microscope, but not in group C. At the same time points, the number of propriocep-tors, the incubation period of SEPs and EMG as well as the wave amplitude of group A was significantly bigger, shorter and higher than group B and C. However, no significant differences were ob-served in all the measurements between group B and C. Conclusion Adenovirus-mediated h NT-3 genetransfection can promote the regeneration of proprioceptors in the autologous tendon graft and the recov-ery of the proprioception after the anterior cruciate ligament reconstruction.