Target for treating silicosis and application thereof

A technology of therapeutic target and lung epithelial cells, which is applied in the field of biomedicine to improve the effect of lung damage

Pending Publication Date: 2022-04-15
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, no research has focused on the application of glycolysis in alleviating the development of silicosis

Method used

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  • Target for treating silicosis and application thereof
  • Target for treating silicosis and application thereof
  • Target for treating silicosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Establishment of Pulmonary Fibrosis Model Induced by Particulate Dust (CS) in Mice

[0052] Healthy 6-8 week old male C57BL / 6 mice were anesthetized by intraperitoneally injecting 3% pentobarbital sodium 50 mg / kg. Perfuse normal saline or SiO through the non-exposed trachea 2 (3 mg / only) to construct a pulmonary fibrosis model, and recover tissues and alveolar lavage fluid after 7, 28, and 56 days.

[0053] (1) Preparation of pathological slices of lung tissue

[0054] After the mouse lung tissue was fixed in 4% paraformaldehyde solution for 1 week, the tissue was removed for trimming and the residual paraformaldehyde in the tissue was washed away, and gradient dehydration (85% ethanol solution for 2 hours, 95% ethanol solution for 1 After 1 hour, change to a new 95% ethanol solution for another 1 hour, and after 0.5 hours for an absolute ethanol solution, change to a new absolute ethanol solution for an additional 0.5 hour). After dehydration, treat with x...

Embodiment 2

[0080] After HBE cells were treated with CS or ferroptosis inducer Erastin, the cells were collected for QPCR, C11 BODIPY581 / 591 lipid peroxidation fluorescent probe staining, indirect immunofluorescence staining, western and GSH, MDA, Fe 2+ Level determination, the specific steps are as follows:

[0081] For QPCR results, see Figure 10 , After CS treatment, the glycolysis-related genes in HBE cells were up-regulated. The above results indicated that the glycolysis process of lung epithelial cells increased in CS-induced lung fibrosis model in vitro.

[0082] (1) C11 BODIPY 581 / 591 lipid peroxidation fluorescent probe staining: Take out the cells from the incubator, add an appropriate volume of C11 BODIPY 581 / 591 to the cells cultured to a suitable density, incubate in the dark for 30 minutes, discard Add 1ml of 1×PBS to the culture medium to wash the cells twice to remove excess dye, and observe the results with a fluorescence microscope.

[0083] C11 BODIPY 581 / 591 lipid ...

Embodiment 3

[0089] After treatment with ferroptosis inhibitor Ferr-1 and DFO, HBE cells were collected for C11 BODIPY581 / 591 lipid peroxidation fluorescent probe staining, indirect immunofluorescence staining, western and GSH, MDA, Fe 2+ Level determination, the specific steps are as follows:

[0090] C11 BODIPY 581 / 591 lipid peroxidation fluorescent probe staining results see Figure 15 , showed that after administration of ferroptosis inhibitors, the green fluorescence signal intensity in cells was significantly weakened, indicating that CS-induced ferroptosis in HBE cells was inhibited, and the level of lipid peroxidation in cells was down-regulated.

[0091] Fe 2+ , GSH, MDA level determination results see Figure 16 , compared with the CS group, after treatment with ferroptosis inhibitors, the level of GSH in HBE cells increased, while MDA production was associated with Fe 2+ Savings decrease.

[0092] The results of indirect immunofluorescence staining are shown in Figure 17 ,...

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Abstract

The invention relates to a silicosis treatment target and application thereof. The treatment target is glycolysis. By regulating and controlling abnormal glycolysis, inhibiting ferroptosis and EMT of epithelial cells and relieving occurrence and development of silicosis, a new direction is provided for medicine development and treatment means related to the silicosis, it is found that abnormal glycolysis occurs in the occurrence and development process of the silicosis, and a theoretical basis is also provided for a clinical auxiliary diagnosis technology of the silicosis.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a silicosis treatment target and its application. Background technique [0002] Silicosis is a type of occupational disease characterized by extensive pulmonary nodules and pulmonary interstitial fibrosis caused by long-term inhalation of a large amount of dust with a high content of free silica. It is the most common and serious type of occupational disease in my country. Silicosis In the later stage, lung function is seriously affected, and various complications occur, which seriously affects the normal life of patients. At present, the development of silicosis is mainly based on symptomatic treatment, and there are no specific drugs and effective treatment methods in clinical practice, mainly relying on adjuvant treatment, such as the use of targeted drugs to protect macrophages from silica dust damage. Drugs, bronchoalveolar lavage, etc. Currently used drugs such as tetr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7004A61P11/00C12Q1/6883G01N33/68
Inventor 陶莎莎李倩敏杨友静薛莲褚开淼
Owner SUZHOU UNIV
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