微创医学

目的探索将具有报告基因的重组腺病毒载体从下肢效应器沿其支配的脊神经转移至脊髓的途径。方法将携带半乳糖苷酶报告基因(LacZ)的5型重组腺病毒载体(Ad5CMVLacZ)注入到成年Wistar大鼠的腓肠肌(病毒滴度为1.01×105pfu/mL),分别在注射后第3天、7天、14天、21天及28天灌注固定动物,选取腓肠肌、与腰骶神经丛相连的脊髓节段以及对应的脊神经节作为观察对象,用β-半乳糖苷酶免疫组织化学反应鉴定被转染和表达蛋白质的细胞。结果在注射腓肠肌3d后,该肌肉内有被转染及表达产物的细胞;7d后在脊神经节内出现被转染和表达的细胞;14d后在脊髓白质的后索及外侧索内出现上述细胞;21～28d后在脊髓灰质的后角、中间带及前角出现被转染和表达的细胞。结论随注射时间推移,腺病毒表达载体可从下肢骨骼肌沿着受支配的脊神经转移到脊髓,所携带的报告基因也成功地表达产物,故这是一条具有潜在应用价值的微创性基因治疗脊髓疾病的途径。

Objective To explore the possibility of delivery of adenoviral vector carrying Lac Z gene from skeletal muscle to the spinal cord.Methods A dose of 20 μl(1.01×105 pfu/ml)of Ad5 CMV Lac z gene was injected into gastrocnemius of the adult rat,then after 7,10,14,21,and 28 days of the infections,the rats were sacrificed to perform β-galactosidase(β-gal)immunohistochemistry for detecting the transfected and expressed cells in the gastrocnemius,spinal ganglia,and the lumbar part of the spinal cord.Results After 3 days of injection of Ad5 CMV LacZ,the cells in the gastrocnemius were transfected and expressed,then after 7 days and 14 days of injections,the cells in related spinal ganglia were transfected and expressed.Finally,after 28 days of injection,the cells in the white matter and grey matter in the lumbar segments of the spinal cord were transfected and expressed.Conclusion The above datum indicate that the lower limb effector-spinal cord is a reliable pathway for transferring foreign gene carried by adenoviral vector to the spinal cord.

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