腺病毒介导的LacZ基因在大鼠坐骨神经的表达

Adenoviral Vector-mediated Transfer of LacZ Genes into Tat Sciatic Nerve

目的 :观察腺病毒介导的LacZ基因在大鼠坐骨神经雪旺细胞 (Schwanncells ,SCs)的表达。方法 :大鼠坐骨神经内直接注射报告基因LacZ重组腺病毒 (Ad LacZ) ,X gal组织化学染色检测LacZ基因的表达 ,采用LEICAM 5 5 0型图像分析仪对坐骨神经切片X gal染色强弱进行定量评价。结果 :将滴度为 1× 10 8PFU /ml的Ad LacZ病毒液 10 μl注入坐骨神经 2d后即可检测到LacZ的表达 ,7～ 14d表达显著增加 (与 2d组比较P <0 .0 1) ,7d组与 14d组表达量无显著差异 (P >0 .0 5 ) ,2 8d时SCs蓝染又减弱 (与 7d组和 14d组比较P <0 .0 1)。注射生理盐水的对照组X gal染色阴性。结论 :腺病毒介导的LacZ基因可转入活体动物坐骨神经内并高效表达 ,表明腺病毒介导神经营养因子等基因治疗有促进周围神经损伤再生的作用。

Objective: To investigate the expression of LacZ gene in rat sciatic nerve introduced by adenoviral vector. Methods: The efficiency of transfection of Ad-LacZ was determined by X-gal histochemical staining in rat sciatic nerve. Results: 10μl of 1×108 PFU/ml Ad-LacZ were injected into rat sciatic nerve. LacZ positive cells were observed 2 days after injection and were increased in 7～14 days following injection of Ad-LacZ(vs.2 days group P<0.01) . The expression of LacZ gene decreased significantly 28 days following injection of Ad-LacZ(vs.7 and 14 days group P>0.01) . The expression of LacZ gene in nerves injected with physiological saline was not observed. Conclusions: It was concluded that an adenoviral vector could be used to efficiently direct the expression of a foreign gene in SCs of intact rat peripheral nerves. The results suggested that neurotrophic factors could be introduced into SCs by adenoviral vector to promote peripheral nerve regeneratin.