Application of azelaic acid in preparation of medicine for preventing and treating radiation-induced lung injury

Abstract

The invention provides application of azelaic acid in preparation of a medicine for preventing and treating radiation-induced lung injury, and belongs to the technical field of biological medicine. The invention provides an application of azelaic acid in preparation of a medicine for preventing and / or treating radiation-induced pulmonary injury, metabonomics analysis finds that the content of azelaic acid in serum of a mouse subjected to 20Gy gamma-ray single total chest irradiation for 1 month and 4 months is remarkably reduced; cell experiments find that after irradiation, expression of inflammatory factors in lung epithelial cells is increased, epithelial-mesenchymal transition occurs to the cells, and consequences caused by radiation after azelaic acid is given are relieved, and animal experiments find that expression of inflammatory factors in serum of a mouse after irradiation is increased, lung functions are damaged, and consequences caused by radiation after azelaic acid is given are relieved. Azelaic acid is proved to have a potential effect of improving radiation-induced lung injury.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of azelaic acid in preparing drugs for preventing and treating radiation lung injury. Background Art

[0002] Radiation-induced lung injury (RILI) is a common complication of thoracic tumor radiotherapy, which is clinically divided into early radiation-induced pneumonia (Radiation-induced pneumonia) and late radiation-induced pulmonary fibrosis (Radiation-induced pulmonary fibrosis, RIPF). Clinical symptoms include retching, shortness of breath, chest pain, fever, and even respiratory failure and death. Due to the lack of effective treatment for RILI, finding drug intervention targets with diagnostic potential has very important clinical application value for the prevention and treatment of radiation-induced lung injury. Summary of the invention

[0003] The present invention provides the use of azelaic acid in the preparation of a drug for preventing and treating radiation-induced lung injury. After irradiation with gamma rays, azelaic acid has a potential effect in preventing and treating radiation-induced lung injury.

[0004] The purpose of the present invention is to provide the use of azelaic acid in preparing a medicine for preventing and / or treating radiation lung injury.

[0005] In one embodiment of the present invention, the administration of the drug includes cell administration and body administration.

[0006] In one embodiment of the present invention, when cells are administered, the dosage of azelaic acid is not less than 10 μM.

[0007] In one embodiment of the present invention, the azelaic acid is administered in the form of a solution, the solvent is DMSO, and the solution is diluted with culture medium.

[0008] In one embodiment of the present invention, when the drug is administered to the body, the dosage of azelaic acid is not less than 10 mg / kg mouse.

[0009] In one embodiment of the present invention, the azelaic acid is administered in the form of a solution, and the solvent is a mixture of DMSO, PEG300, Tween-80 and ddH2O.

[0010] In one embodiment of the present invention, the solvent comprises, by volume percentage, 3% DMSO, 40% PEG300, 5% Tween-80 and 52% ddH2O.

[0011] Another object of the present invention is to provide a drug for preventing and / or treating radiation lung injury, the active ingredient of which includes azelaic acid and pharmaceutically acceptable excipients.

[0012] Beneficial effects: The present invention provides the use of azelaic acid in the preparation of drugs for preventing and / or treating radiation lung injury. Animal experiments have shown that after a single whole-chest local irradiation with 20Gy γ rays, azelaic acid is administered by intraperitoneal injection at a rate of 10 mg / kg body weight once every three days. Metabolomics analysis has shown that the azelaic acid content in the serum of mice 1 month and 4 months after a single whole-chest irradiation with 20Gy γ rays was significantly reduced. Cell experiments have shown that 10μM and 20μM cells are administered before 20Gy γ ray irradiation and cultured for 48h after 20Gy γ ray irradiation. Studies have shown that the expression of inflammatory factors in cells increases after irradiation, and the cells undergo epithelial-mesenchymal transition. After the administration of azelaic acid, the consequences of radiation are alleviated, confirming that azelaic acid has the potential to improve radiation lung injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is a graph showing the changes in coat color in mice;

[0014] Figure 2 This is a graph showing the weight changes of mice;

[0015] Figure 3 This is the graph of lung coefficient changes;

[0016] Figure 4 This is the respiratory function index change chart;

[0017] Figure 5 This is a graph of physiological changes in mice when samples were taken on day 120;

[0018] Figure 6 HE staining, Masson staining and quantitative results when sampling at different times;

[0019] Figure 7 This is the change chart of serum inflammatory factors IL-1β and IL-6;

[0020] Figure 8 This is a graph showing the results of inflammatory factors in lung epithelial cells caused by ionizing radiation;

[0021] Fig. 9 This is a diagram showing the epithelial-mesenchymal transition of lung epithelial cells caused by ionizing radiation;

[0022] Fig.10 A diagram showing the effect of azelaic acid on improving ionizing radiation-induced lung fibrosis in mice. DETAILED DESCRIPTION

[0023] The purpose of the present invention is to provide the use of azelaic acid in preparing a medicine for preventing and / or treating radiation lung injury.

[0024] The azelaic acid of the present invention has a CAS number of 123-99-9 and a molecular formula of C9H 16 O4; In this field, azelaic acid is a non-toxic, non-teratogenic, pure natural nine-carbon straight-chain saturated acid. In 1978, dermatologist Marcella Nazzaro-Porro first reported that it was extracted from a culture medium of Malassezia. Since the 21st century, research on azelaic acid has mainly focused on its chemical structure and research on skin diseases such as acne and rhinitis. The present invention uses the azelaic acid to prevent and treat radiation lung injury.

[0025] In the examples of the present invention, the preventive and therapeutic effects of azelaic acid on radiation-induced lung injury were verified by cell experiments and animal experiments.

[0026] In one embodiment of the present invention, especially in cell experiments, the dosage of azelaic acid is not less than 10 μM, and the administration is in the form of a solution, the solvent is DMSO, and the solution is diluted with culture medium. In this embodiment of the present invention, first, 11.2932 mg of azelaic acid solid is dissolved in 1 mL of DMSO to prepare a 60 mM azelaic acid mother solution, and 10 μL of the 60 mM mother solution is dissolved in 50 μL of DMSO to obtain a 10 mM working solution, and 10 μL of the 60 mM mother solution is dissolved in 20 μL of DMSO to obtain a 20 mM working solution. During the experiment, 10 μM and 20 μM azelaic acid solutions are prepared according to the volume ratio of working solution: culture medium = 1:1000.

[0027] In one embodiment of the present invention, especially in animal experiments, the dosage of azelaic acid is not less than 10 mg / kg mouse. And the azelaic acid is administered in the form of a solution, and the solvent (Vehicle) is a mixture containing DMSO, PEG300, Tween-80 and ddH2O. In a specific embodiment, the solvent contains, by volume percentage: 3% DMSO, 40% PEG300, 5% Tween-80 and 52% ddH2O. In the specific configuration, first take 0.04 mg of azelaic acid (AZA) and dissolve it in 1 μL DMSO to obtain a mother solution of 0.04 mg / μL; then configure it according to the following volume ratio: 3% azelaic acid mother solution, 40% PEG300, 5% Tween-80 and 52% ddH2O.

[0028] Another object of the present invention is to provide a drug for preventing and / or treating radiation lung injury, the active ingredient of which includes azelaic acid and pharmaceutically acceptable excipients.

[0029] In order to further illustrate the present invention, the application of azelaic acid provided by the present invention in the preparation of drugs for preventing and treating radiation lung injury is described in detail below in combination with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0030] Example 1

[0031] The experiment was divided into three groups:

[0032] NC+Vehicle group: 7-week-old male C57BL / 6J mice were purchased and bred in an SPF environment for one week, and then injected intraperitoneally with the solvent at 10 mg / kg body weight every three days;

[0033] IR+Vehicle group: 7-week-old male C57BL / 6J mice were purchased and adapted to SPF environment for one week, then irradiated with 20Gy gamma rays to the whole chest once, and then injected with solvent intraperitoneally at 10mg / kg body weight once every three days;

[0034] IR+AZA group: Seven-week-old male C57BL / 6J mice were purchased and adaptively raised in an SPF environment for one week. After that, they were irradiated with 20Gy γ-rays on the whole chest once. Then, azelaic acid was intraperitoneally injected at 10mg / kg body weight once every three days.

[0035] Solvent (Vehicle): 3% DMSO + 40% PEG300 + 5% Tween-80 + 52% ddH2O;

[0036] Azelaic acid reagent (AZA): 3% azelaic acid stock solution + 40% PEG300 + 5% Tween-80 + 52% ddH2O;

[0037] Azelaic acid mother solution: Dissolve 0.04 mg of AZA in 1 μL of DMSO to obtain a 0.04 mg / μL mother solution.

[0038] Experimental process:

[0039] 1. Sample collection in 15 days

[0040] NC+Vehicle group: after one week of adaptive feeding, the solvent was intraperitoneally injected five times every three days;

[0041] IR+Vehicle group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and the solvent was given immediately after irradiation, once every three days, for a total of 5 intraperitoneal injections;

[0042] IR+AZA group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and AZA was given immediately after irradiation, once every three days, for a total of 5 intraperitoneal injections;

[0043] Samples were collected every 14 days during the experiment to observe the coat color, body weight, lung coefficient, and HE and Masson staining results of lung tissue.

[0044] 2. 30 days to collect samples

[0045] NC+Vehicle group: after one week of adaptive feeding, the solvent was intraperitoneally injected 10 times every three days;

[0046] IR+Vehicle group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and the solvent was given immediately after irradiation, once every three days, for a total of 10 intraperitoneal injections;

[0047] IR+AZA group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and AZA was given immediately after irradiation, once every three days, for a total of 10 intraperitoneal injections;

[0048] Samples were collected every 30 days during the experiment to observe the coat color, body weight, lung coefficient, and HE and Masson staining results of the lung tissues of the mice.

[0049] Weight changes such as Figure 2 As shown, the weight of mice decreased after irradiation, and azelaic acid had no effect on improving the weight loss caused by ionizing radiation;

[0050] Lung coefficient changes as Figure 3 As shown, the lung coefficient of mice increased after irradiation, which was relieved after administration of azelaic acid;

[0051] Respiratory function index changes such as Figure 4 As shown, PIF decreased and Penh increased in mice after irradiation, which was relieved after administration of azelaic acid.

[0052] 3. 120 days to collect samples

[0053] NC+Vehicle group: After one week of adaptive feeding, the mice were intraperitoneally injected with solvent 10 times for a total of 30 days, and then fed normally for 90 days;

[0054] IR+Vehicle group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and the solvent was immediately given intraperitoneally 10 times for a total of 30 days, and then the mice were fed normally for 90 days;

[0055] IR+AZA group: After one week of adaptive feeding, the whole chest was irradiated with 20Gy gamma rays once, and AZA was given intraperitoneally 10 times immediately after irradiation, for a total of 30 days, and then fed normally for 90 days;

[0056] Samples were collected after 120 days of the experiment to observe the coat color, body weight, lung coefficient, and HE and Masson staining results of lung tissue.

[0057] The coat color of the 120-day-old mouse model is as follows Figure 1 As shown, the chest hair of mice turned white and fell off after irradiation, and hair loss in the area was alleviated after administration of azelaic acid;

[0058] like Figure 5 and Figure 6 As shown in the figure, the mice lost weight after irradiation; the lung coefficient of mice increased after irradiation, which was relieved after administration of azelaic acid; the PIF of mice decreased after irradiation, and the Penh increased, which was relieved after administration of azelaic acid; after 120 days of observation, it was found that azelaic acid had no significant effect on the weight loss caused by irradiation, but the administration of azelaic acid at this time showed an improvement trend for the decrease in the lung coefficient. Changes in serum inflammatory factors IL-1β and IL-6 Figure 7 As shown, both increased after irradiation and improved after administration of azelaic acid.

[0059] Mouse lung tissue protein was extracted for Western Blot experiment. The results are as follows Fig.10 As shown, the same results were found as in the cell experiments, namely, azelaic acid improved the mesenchymal transformation of lung epithelial cells induced by ionizing radiation.

[0060] The above results showed that mice developed radiation pneumonia 14 days and 30 days after irradiation, and radiation pulmonary fibrosis 120 days after irradiation, and the damage was improved after administration of azelaic acid reagent.

[0061] Example 2

[0062] Cell experiment: Azelaic acid stock solution (60 mM): 11.2932 mg of azelaic acid solid was dissolved in 1 mL of DMSO;

[0063] Azelaic acid working solution (10 mM, 20 mM): Dissolve 10 μL of 60 mM stock solution in 50 μL DMSO to obtain a 10 mM working solution. Dissolve 10 μL of 60 mM stock solution in 20 μL DMSO to obtain a 20 mM working solution.

[0064] During the experiment, 10μM and 20μM azelaic acid solutions were prepared at a ratio of working solution: culture medium = 1:1000. 10μM and 20μM azelaic acid were given to lung epithelial cells (A549, MLE12) immediately before 10Gy γ-ray irradiation. After irradiation, samples were collected and cultured for 48 hours to observe the changes in cellular inflammatory factors IL-1β and IL-6 and the cell epithelial-mesenchymal transition.

[0065] Azelaic acid can improve the increase of inflammatory factors in lung epithelial cells caused by ionizing radiation ( Figure 8 ); Azelaic acid can improve the epithelial-mesenchymal transition of lung epithelial cells induced by ionizing radiation ( Fig. 9 ).

[0066] The above results indicate that the expression of inflammatory factors in cells increased after irradiation, and the cells underwent epithelial-mesenchymal transition, while the above radiation consequences were alleviated after the administration of azelaic acid.

[0067] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Use of azelaic acid in the preparation of drugs for preventing and / or treating radiation lung injury.

2. The application according to claim 1, characterized in that: The administration methods of the drug include cell administration and body administration.

3. The application according to claim 2, characterized in that: When the cell is administered, the dosage is not less than 10 μM in terms of azelaic acid dosage.

4. The use according to claim 3, characterized in that: The azelaic acid was administered in the form of a solution, the solvent was DMSO, and the solution was diluted with culture medium.

5. The use according to claim 1, characterized in that: When administered to the body, the dosage of azelaic acid is not less than 10 mg / kg mouse.

6. The use according to claim 5, characterized in that: The azelaic acid is administered in the form of a solution, wherein the solvent is a mixed solution containing DMSO, PEG300, Tween-80 and ddH2O.

7. The use according to claim 6, characterized in that: The solvent contains, by volume percentage, 3% DMSO, 40% PEG300, 5% Tween-80 and 52% ddH2O.

8. A drug for preventing and / or treating radiation-induced lung injury, characterized in that: The active ingredient comprises azelaic acid and also comprises pharmaceutically acceptable excipients.

Images