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Application of guanosine triphosphate cyclohydrolase 1 and drug

A technology of guanosine triphosphate and hydrolase, which is applied in the biological field to achieve the effect of enhancing cell proliferation ability

Active Publication Date: 2019-10-22
SUZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of GCH1 in the prevention and treatment of radiation-induced lung injury has not been seen so far at home and abroad, and its mitigating effect on radiation injury needs more research and exploration

Method used

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  • Application of guanosine triphosphate cyclohydrolase 1 and drug
  • Application of guanosine triphosphate cyclohydrolase 1 and drug
  • Application of guanosine triphosphate cyclohydrolase 1 and drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Construction of GCH1 overexpression adenovirus and expression verification after infection

[0035] In this example, the whole gene synthesis method (Shandong Weizhen Biotechnology Co., Ltd.) was used to synthesize the human GCH1 coding region gene (the nucleic acid sequence is shown in Table 1 below), and the target fragment was cloned into the AdTrack-CMV shuttle plasmid, and the adenovirus backbone was used to Plasmid pAdeasy-1 was subjected to homologous recombination in BJ5183 bacteria, and positive clones were selected to transfect 293A cells with small plasmids. The pAd-GCH1 (target adenovirus) and pAd-NC (control adenovirus) recombinant adenoviruses were obtained through packaging.

[0036] Table 1 The sequences of primers and genes

[0037]

[0038]

[0039] Human embryonic lung fibroblasts HELF in the exponential growth phase were mixed with 10.0×10 4 Cells / well were seeded in a 6-well plate, and 2 μL of 1×10 12 vp / ml GCH1 overexpression ade...

Embodiment 2

[0040] Example 2 Anti-radiation effect test of GCH1 overexpression adenovirus on lung cells

[0041] 1. GCH1 overexpression adenovirus reverses NO reduction induced by ionizing radiation

[0042] Human embryonic lung fibroblasts in exponential growth phase were seeded in 96-well plates and cultured for 12 hours, and Ad-NC or Ad-GCH1 adenovirus was added 24 hours before irradiation, respectively. Each group had 4 parallel samples, and the experiment was repeated 3 times. Afterwards, X-ray irradiation with an absorbed dose of 20Gy was given. Aspirate the medium 24 hours after exposure, add 100 μL of diluted DAF-FM DA to each well (dilute DAF-FM DA with DAF-FM DA diluent according to 1:1000, so that the final concentration is 5 μmol / L), and place in the incubator Incubate for 20min. Wash three times with PBS to fully remove DAF-FMDA that did not enter the cells. Detection by microplate reader.

[0043] Such as figure 2 As shown (P<0.05), the results showed that the GCH1 ov...

Embodiment 3

[0050] Example 3 GCH1 overexpression adenovirus anti-radiation effect test on mouse lung tissue

[0051] Male C57 mice aged 6 to 8 weeks were purchased from Shanghai Slack Experimental Animal Co., Ltd., and each mouse was anesthetized by intraperitoneal injection of 4% chloral hydrate according to 1% of its body weight, and small animal positioning was performed using a small animal precision irradiation device Afterwards, the right lung was precisely irradiated with an aperture of 5mm, so that 100% of the volume reached the prescribed dose of 20Gy, and a mouse radiation-induced lung injury model was established, and then the mice were randomly divided into 2 groups (5 in each group):

[0052] 1) Give mice a tail vein injection of 100 μL of 1×10 12 vp / ml GCH1 overexpression adenovirus; 2) inject 100 μL of 1×10 12 vp / ml GCH1 control adenovirus (Ad-NC).

[0053] After 7 days of irradiation, C57 mice were sacrificed by cervical dislocation. The right lung tissue was taken, rin...

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Abstract

The invention relates to application of guanosine triphosphate cyclohydrolase 1 (GCH1) and a drug. It is found that GCH1 overexpression has the prevention and treatment effect on radiation injuries ofhuman lung cells and animal model lung tissues, and can enhance oxidation resistance of the cells and cell proliferation capacity after radiation. A GCH1 overexpression adenovirus or the drug containing the GCH1 overexpression adenovirus can be used for preventing and treating human acute lung radiation injuries, and has the important significance and application value in coping with nuclear andradiation accidents and nuclear emergencies, reducing the lung radiation injury effects on tumor radiation patients and the like.

Description

technical field [0001] The invention relates to the application and medicine of guanosine triphosphate cyclohydrolase 1, belonging to the field of biotechnology. Background technique [0002] Radiation-induced lung injury is a common complication after radiotherapy for thoracic tumors and pretreatment for bone marrow transplantation. In addition, since the lung is a radiation-sensitive organ, occasional nuclear radiation accidents in normal times or nuclear radiation damage in wartime often involve the lungs, resulting in radiation-induced lung injury. In tumor radiotherapy, a considerable number of patients will have different degrees of radiation-induced lung injury. Radiation lung injury is more serious in patients with thoracic malignancies, such as lung, breast and esophageal cancer. Severe radiation-induced lung injury may affect the smooth progress of radiotherapy and the quality of life of patients. [0003] Radiation-induced lung injury refers to a series of path...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/50A61K48/00A61P11/00
CPCA61K38/50A61K48/0025A61K48/005A61P11/00C12Y305/04016
Inventor 曹建平张镇李旭刚冯阳高晶晶张舒羽聂继华
Owner SUZHOU UNIV
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