Use of recombined adenovirus carrying hepatocyte growth factor gene

A technology of hepatocyte growth factor and recombinant adenovirus, which is applied in the field of biomedicine, can solve medical problems, and there are no effective means for the prevention and treatment of fibrous diseases, etc., and achieve a clear effect

Inactive Publication Date: 2004-03-17
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] To sum up, there is no effective method for the prevention and tre

Method used

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  • Use of recombined adenovirus carrying hepatocyte growth factor gene
  • Use of recombined adenovirus carrying hepatocyte growth factor gene
  • Use of recombined adenovirus carrying hepatocyte growth factor gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Construction, amplification and purification of recombinant adenovirus 1. Construction of recombinant adenovirus

[0033] The construction process of the shuttle plasmid (pXCJL1-CMV / HGF / polyA) carrying the human hepatocyte growth factor gene is as follows: figure 1 shown. 293 cells were co-transfected with pXCJL1-CMV / HGF / polyA and the recombinant plasmid GT4050 carrying the genomic DNA of the adenovirus, and homologous recombination occurred in the cells to form a recombinant adenovirus carrying the human hepatocyte growth factor gene (named as Ad -HGF), the schematic diagram of its structure is shown in figure 2 shown. 2. Preparation and purification of recombinant adenovirus

[0034] 293 cells were inoculated in a 650ml cell culture flask. When the cells grew to 90% confluence, the virus was added in an amount of 10 plaque-forming units (pfu) / cell. After 36-48 hours, the cells showed a complete pathological effect ( CPE), collect the cells and fre...

Embodiment 2

[0035] Example 2 Recombinant adenovirus mediates human hepatocyte growth factor gene

[0036]Evaluation of rat liver fibrosis treatment 1. Preparation of rat liver fibrosis model

[0037] Thirty Wistar male rats, weighing 200-250 grams. 26 rats were randomly selected to prepare a fibrosis model: 1 g of dinitronitrosamine (DMN) was dissolved in 100 ml of sterile saline to make the final concentration 1%. Rats were administered subcutaneously, 1ml / kg body weight each time, twice a week, for 4 consecutive weeks; the other 4 rats were used as the normal group, and the other conditions were the same as the model group except that no drug was administered. Ten days after the last administration, two rats in each of the model group and the normal group were randomly selected, and the liver tissues were taken for HE section staining to observe the pathological changes in the liver of the rats with liver fibrosis; liver pathological examination: the rats in the model group The...

Embodiment 3

[0039] Example 3 Recombinant adenovirus-mediated human hepatocyte growth factor gene

[0040] Preventive effect on scar formation 1. Scar animal model

[0041] New Zealand female white rabbits, weighing 2.5-3.5kg. After shaving and disinfecting the avascular area on the ventral side of the ear, a trephine was used to microsurgically remove the full-thickness skin until the surface of the cartilage. The diameter of the circular incision was 0.6 cm, and there were 4-6 incisions on each side. 2. Effect evaluation

[0042] Apply the recombinant adenovirus to the wound surface while excising the skin, 8ul each time (8.6×10 7 pfu / μl) (Ad-HGF group), once a day for 3 consecutive days; the wounds of the control group were smeared with the same dose of Ad-GFP. On the 22nd day after the operation, the wound and the adjacent non-wound skin were excised, paraffin sections were made, and HE staining was performed. The results showed that the Ad-HGF application ...

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Abstract

The present invention relates to biomedicine. Human hepatocyte growth factor full-length code area cDNA is inserted to defect duplicated adenovirus vector GT4050 to obtain recombinant adenovirus with carried human hepatocyte growth factor. When it is injected to rat body, the recombinant adenovirus has treating effect on rat's liver fibrillation. When it is applied to wound, the recombinant adenovirus can prevent scar.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of a recombinant adenovirus carrying human hepatocyte growth factor gene in the preparation of drugs for preventing and treating fibrous diseases. Background technique [0002] Fibrotic diseases, such as liver fibrosis, pulmonary fibrosis, renal fibrosis and scar formation, are one of the diseases that seriously affect human life and health. Liver fibrosis is one of the most important pathological features of chronic liver diseases, and it is also one of the main reasons for the further development of diseases such as chronic viral hepatitis, metabolic disorders or chronic alcoholism. At present, the treatment of liver fibrosis in China only stays in delaying and blocking the process of fibrosis, and for the already formed liver fibrosis, there is no effective means to reverse the process of fibrosis and achieve the effect of treating liver fibrosis. Although there are...

Claims

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Application Information

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IPC IPC(8): C07K14/475C12N7/01C12N15/861
CPCC07K14/4753C12N15/86C12N2710/10343C12N2800/30A61P1/16
Inventor 哈小琴吴祖泽郭树华
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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