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Application of excited GSTP1 gene in medicine for preventing radioactive lung injury

A radiation-induced lung injury and drug technology, applied in the direction of drug combinations, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the problems of no one reporting, etc., to reduce the damage and improve the effect of radiation-induced lung injury

Pending Publication Date: 2021-06-04
THE FIRST MEDICAL CENT CHINESE PLA GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the application of reagents that enhance GSTP1 gene expression in the preparation of drugs for the prevention and treatment of ionizing radiation-induced radiation-induced lung injury has not been reported at home and abroad.

Method used

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  • Application of excited GSTP1 gene in medicine for preventing radioactive lung injury
  • Application of excited GSTP1 gene in medicine for preventing radioactive lung injury
  • Application of excited GSTP1 gene in medicine for preventing radioactive lung injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] GSTP1 overexpression plasmid can significantly enhance GSTP1 protein in normal human lung epithelial cells:

[0025] experiment procedure:

[0026] (1) Cell culture: Culture BEAS-2B cells in LHC-8 medium and add penicillin at a final concentration of 100 U / mL and streptomycin (double antibody) at 100 μg / mL; place the cells at 37°C for 5 Culture in % CO 2 incubator, subculture once every 2-3 days, and take logarithmic growth phase cells for experiments.

[0027] (2) Cell transfection: BEAS-2B was inoculated in a 35 mm diameter petri dish, each dish was inoculated with 3×10 5 cells, and cultured for 24 hours. Avoid antibiotics during plating and transfection. Preparation of transfection solution: The dosage ratio of plasmid: Lipofectamine2000 is 4 μg / well: 10 μl / well. Add the prepared transfection solution into the BEAS-2B cell culture dish to be transfected, and place the culture dish in an incubator for incubation. After 6 hours, the culture medium containing the tr...

Embodiment 2

[0032] The expression of GSTP1 had no effect on cell proliferation. Under radiation stress, the cell proliferation activity of the overexpression group was higher than that of the empty plasmid transfection group (P<0.05):

[0033] experiment procedure:

[0034] (1) Cell culture and cell transfection are the same as in Example 1;

[0035] (2) MTT experiment method: Digest and count BEAS-2B cells in the logarithmic growth phase, resuspend the cells to make the cell concentration 6x104 cells / ml, absorb 200 μl of the cell suspension, and inoculate the cells into a 96-well plate, 37 ℃ incubator incubation. Add 20 μl of working concentration MTT solution (0.5 mg / ml) to each well of the 96-well plate, incubate in the incubator for 4 hours, and then take it out. Discard the culture medium, add 150 μl DMSO to each well, and use a microplate reader (490 nm wavelength) to detect after 10 min.

[0036] Experimental results:

[0037] by right figure 2The growth and proliferation cha...

Embodiment 3

[0039] Enhance the expression of GSTP1 gene, inhibit the occurrence of cell EMT process caused by radiation and weaken the cell damage caused by radiation:

[0040] experiment procedure:

[0041] (1) Cell culture and cell transfection are the same as in Example 1;

[0042] (2) Take BEAS-2B cells transfected with GSTP1 overexpression plasmid and negative control plasmid for 48 hours in the logarithmic growth phase, extract protein, and perform Western blot electrophoresis. Expose the bands of GSTP1, N-cadherin, Vimentin, E-cadherin and β-ACTIN using an exposure instrument.

[0043] Among them, the irradiation conditions: 60Coγ-ray irradiation of the Academy of Military Sciences. Cells were treated with 6Gy of γ-ray irradiation at a dose rate of 1Gy / min.

[0044] Experimental results:

[0045] Statistical processing: All the experiments in the above examples were repeated more than 3 times, and SAS statistical software was used to perform t-test on the relevant data, and P<0...

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Abstract

The invention relates to the field of gene medicines, and particularly relates to application of an excited GSTP1 gene in a medicine for preventing radioactive lung injury. Firstly, GSTP1 over-expression plasmids are verified to be capable of remarkably enhancing GSTP1 protein in human normal lung epithelial cells. Furthermore, an MTT experiment is used for finding that over-expression GSTP1 has no influence on cell proliferation, and under radiation pressure, the cell proliferation activity of an over-expression group is higher than that of a no-load plasmid transfection group. Finally, a Westernblot experiment is used for obtaining enhanced GSTP1 gene expression, the occurrence of a cell EMT process caused by radiation is inhibited, and cell injury caused by radiation is weakened. The invention provides the GSTP1 gene and novel application of a reagent for enhancing GSTP1 gene expression. By enhancing GSTP1 gene expression, the injury of radiotherapy to normal lung tissue can be weakened, and therefore the radioactive lung injury caused by ionizing radiation can be effectively prevented.

Description

technical field [0001] The invention relates to the field of gene medicine, in particular to the application of activating GSTP1 gene in medicine for protecting radiation-induced lung injury. Background technique [0002] Studies in recent years have shown that lung cancer is the malignant tumor with the highest morbidity and mortality in my country. With the development of radiotherapy technology and equipment, radiotherapy has become an indispensable key method for the treatment of lung cancer. The basic principle of radiation therapy is to fully irradiate the lesion with high-energy rays, thereby directly damaging DNA or mediating intracellular water molecules to generate a large amount of reactive oxygen species to damage DNA, causing DNA breaks that cannot be repaired by the cells themselves, and promoting tumor cell death. However, studies have shown that due to relevant limitations such as irradiation techniques, most patients will experience varying degrees of norma...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61P11/00A61P39/00A61P17/16
CPCA61K45/06A61P11/00A61P39/00A61P17/16
Inventor 曲宝林杜乐辉雷霄贺琦多俞伟马娜刘芳黄祥饶乐王瑶梁延杰
Owner THE FIRST MEDICAL CENT CHINESE PLA GENERAL HOSPITAL
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