| LIU Chunjie,WANG Dewen,ZHANG Zhaoshan.Preliminary studies on gene therapy with TGF β1 antisense gene/liposome complexes and adenoviral transfer vector in RPF rats[J].Chinese Journal of Radiological Medicine and Protection,2001,21(4):282-285 |
| Preliminary studies on gene therapy with TGF β1 antisense gene/liposome complexes and adenoviral transfer vector in RPF rats |
| Received:August 20, 2000 |
| DOI： |
| KeyWords:Rat Transforming growth factor-beta l Lung fibrosis Gene therapy |
| FundProject:全军"九·五"指令性基金资助项目(96L011) |
| Author Name | Affiliation | | LIU Chunjie | Beijing Institute of Radiation Medicine, Beijing 100850, China | | WANG Dewen | Beijing Institute of Radiation Medicine, Beijing 100850, China | | ZHANG Zhaoshan | 北京生物工程研究所 | | 高亚兵 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 熊呈琦 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 龙建银 | 北京基础医学研究所 | | 王会信 | 北京基础医学研究所 | | 彭瑞云 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 崔雪梅 | Beijing Institute of Radiation Medicine, Beijing 100850, China |
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| Abstract:: |
| Objective To observed the efficiency of gene therapy with TGFβ1 antisense gene/liposome complexes and adenoviral transfer vector in RPF rats. Methods TGFβ1 sense and antisense gene expression vectors and adenovirus transfer vector were introduced into rat bronchus by way of intratracheal instillation. Results At day 1.5 after TGFβ1 sense and antisense gene transfer,PCR amplification using neo gene-specific primer from lung tissue DNA was all positive.After day 5.5,67%(2/3) of lung tissue DNA was positive.RNA dot blot hybridization indicated that TGFβ1 mRNA content of lung tissue transfected with pMAMneo-AntiTGFβ1 gene decreased.Detection of lung hydroxyproline (Hyp) content after day 35 of gene transfer showed that even in lung of rats received pMAMneo-AntiTGFβ1 lipid complexes it raised remarkably (P <0.01) compared with that of normal animals;it was lower singnificantly (P <0.01) than that of the rats received pMAMneo-TGFβ1 lipid complexes and of the rats received saline treatment.The replication deficient recombinant adenovirus bearing firefly luciferase gene were purified by cesium chloride density centrifugation.The virus of 2.5×109 pfu/ml were instilled into bronchus at 0.5 ml per rat.After day 2 and day 6,the lung tissues of all six rats (three per each group) expressed the transfected luciferase gene by luminometer. Conclusion Cationic lipid mediated TGFβ1 antisense gene therapy was a simple and easy method.It can slow down the course of pathogenesis of lung fibrosis.Replication deficient recombinant adenovirus mediated gene therapy of lung diseases is a good and efficient method. |
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