摘要
目的 观察以腺病毒为载体的外源性治疗基因Ad CMV hTGF β1转染兔椎间盘髓核细胞后的表达。 方法 采用本实验室构建的腺病毒载体基因Ad CMV hTGF β12 0 μl(6× 10 6 pfu)注射于纯种成年新西兰大白兔腰椎间盘髓核内 ;手术后不同时间分别取出椎间盘 ,用免疫组织化学方法对椎间盘髓核细胞进行基因表达产物测定 ;用VIDAS图像分析系统进行半定量观察。 结果 免疫组织化学染色显示注射Ad CMV hTGF β1椎间盘 ,4d组可见髓核细胞内呈现阳性染色颗粒 ;1周组即显示强阳性染色 ,阳性表达持续至 12周组。实验对照组及自身椎间盘对照组呈阴性或弱阳性反应。 结论 椎间盘髓核作为靶细胞能够被外源性基因转染 ;腺病毒载体基因Ad CMV hTGF β1转染兔椎间盘髓核细胞后能在相当长时间内表达TGF
Objective To deliver an exogenous therapeutic gene—hTGF β1 to rabbit lumbar intervertebral disc in vivo by adenovirus vector and observe the expression of the exogenous therapeutic gene. Methods 20μl of 0 01?mol/L PBS, with or without adenovirus (6×10 6pfu) carrying TGF β1 gene (Ad/CMV hTGF β1) which were constructed by our laboratory, was injected directly into nucleus pulposus tissues of lumber discs of mature New Zealand white rabbits. Immunohistochemical staining for human transforming growth factor β1 was performed on the rabbit disc tissues in different periods after operation. A half quantitation for immunohistochemical staining was performed by VIDAS analysis system. Results Discs injected with Ad/CMV hTGF β1 exhibited extensive and intense positive immunohistochemical staining for transforming growth factor β1 from 1 week to 12 weeks after operation, and positive pellets in nucleus pulposus cells in 4th day group, while the control groups(intact group and PBS group) showed negative or weakly positive immunohistochemical staining. The OPTDM was significantly increased in the discs injected with Ad/CMV hTGFβ1 compared to the contact discs or the discs injected with only PBS( P <0 05).Conclusion The nucleus pulposus tissues of the rabbit discs can be transferred by the adenovirus mediated vector carrying the human transforming growth factor β1 enconding gene, and can effectively express the hTGF β1 protein in a long period.
出处
《解剖学报》
CAS
CSCD
北大核心
2003年第1期63-66,共4页
Acta Anatomica Sinica
基金
山东省卫生厅"九五"攻关资助项目 [鲁卫科教字 (1996)第 12号 ]
