Medicine for preventing and controlling radiation skin injuries and application thereof
A radioactive skin and drug technology, applied in the prevention and treatment of radioactive skin damage, in the field of fatty acid binding protein 4 overexpression adenovirus and/or FABP4 drug, can solve the problems affecting the quality of life of radiotherapy patients, microvascular and small blood vessel endothelial cell damage, Necrosis and other problems, achieve the effect of enhancing anti-oxidation and DNA damage repair ability, maintaining skin barrier function, and inhibiting free radical oxidation
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Embodiment 1
[0037] Example 1, construction of FABP4 overexpression adenovirus and expression verification after infection
[0038]The whole gene synthesis method (synthesized by Shanghai Sangon Bioengineering Co., Ltd.) was used to synthesize the human FABP4 coding region gene (the nucleic acid sequence is shown in SEQ ID NO.1), and the target fragment was cloned into the AdTrack-CMV shuttle plasmid, and the adenovirus was used to The backbone plasmid pAdeasy-1 was subjected to homologous recombination in BJ5183 bacteria, and the positive clones were selected to transfect 293A cells with small plasmids. The pAd-FABP4 (target adenovirus) and pAd-NC (control adenovirus) recombinant adenoviruses were obtained through packaging.
[0039] Skin fibroblast WS1 cells in exponential growth phase were treated with 2.0×10 4 Cells / well were seeded in a 24-well plate, and 1×10 5 FABP4 overexpression adenovirus or control adenovirus (Ad-NC), at 37°C, 5% CO 2 Continue culturing for 24 h in the incuba...
Embodiment 2
[0041] Example 2, FABP4 overexpression adenovirus anti-radiation effect test on skin cells
[0042] 1. FABP4 overexpression adenovirus reduces radiation-induced free radicals
[0043] Skin fibroblast WS1 cells in exponential growth phase were treated with 2.0×10 4 The density of cells / well was inoculated in 24-well plates, and after the cells adhered to the wall, 1×105 FABP4 overexpressed adenovirus or control adenovirus (Ad-NC) was added to each well, and the cells were incubated at 37°C and 5% CO 2 Continue culturing for 24 h in the incubator. Each group had 4 parallel samples, and the experiment was repeated 3 times. Then give the absorbed dose of 5GyX-ray irradiation. Aspirate the medium 1 hour after exposure, add 1 mL of diluted DCFH-DA to each well (dilute DCFH-DA with serum-free medium according to 1:1000, so that the final concentration is 10 μmol / L), and incubate in the incubator for 20 minutes. Wash three times with serum-free medium to fully remove DCFH-DA that ...
Embodiment 3
[0048] Embodiment 3, FABP4 overexpression adenovirus anti-radiation effect test on animal skin tissue
[0049] Male SD rats were purchased from Shanghai Slack Experimental Animal Co., Ltd., and each rat was anesthetized by intraperitoneal injection of 4% chloral hydrate according to 1% of its body weight. Unintentional movement during irradiation. The buttock skin of rats was irradiated with 6MeV electron beams produced by the 23EX linear accelerator of Varian Corporation of the United States. The surface was covered with a 1cm-thick tissue compensation film, and the non-irradiated parts were shielded with lead plates. The dose was 750cGy / min, the dose was 45Gy, and the irradiation area was outlined with a marker pen before irradiation, and then the rats were randomly divided into 2 groups (5 rats in each group): 1) subcutaneous injection of 1ml of 1×109FABP4 to the rats Expression of adenovirus; 1) subcutaneous injection of 1ml of 1×10 9 FABP4 control adenovirus (Ad-NC).
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