Application of irisin in preparation of medicine for treating radiation-induced lung injury
By applying drugs prepared by irisin, the problem of poor efficacy of radioactive lung injury was solved, which significantly alleviated radiopneumonia and fibrosis, reduced inflammatory factors release and oxidative stress, enhanced lung tissue barrier function, and provided new therapeutic targets.
Patent Information
- Application Number
- CN202510619847.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-14
- Publication Date
- 2025-07-11
AI Technical Summary
The prior art has poor efficacy and great side effects in the treatment of radioactive lung injury. How to effectively alleviate symptoms such as radiopneumonia and pulmonary fibrosis is a major clinical issue.
Irisin is used to prepare drugs for treating radioactive lung injury, and the dose is 1μg/kg-10μg/kg, supplemented by pharmaceutically acceptable auxiliary materials, which are used to alleviate radiation-induced pulmonary edema, thickening of alveolar interstitial, congestion, alveolar collapse, thickening of the airway wall, narrowing of the airway cavity and collagen deposition around the airway, reduce the release of inflammatory factors and oxidative stress, and enhance the function of lung tissue barrier.
Irisin significantly reduces the damage to lung tissue caused by radiation, reduces the synthesis and release of inflammatory factors, inhibits oxidative stress, reduces the degree of lung fibrosis, restores lung tissue function, and provides new therapeutic targets.
Smart Images
Figure CN120285157A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to the application of irisin in the preparation of drugs for treating radiation-induced lung injury. Background Art
[0002] Radiation-induced lung injury is a serious complication of radiotherapy (RT), usually occurring after radiotherapy for chest tumors. This injury includes two main forms: early radiation pneumonitis (RP) and late radiation fibrosis (RF). Its clinical manifestations are cough, dyspnea, reduced lung function, and in severe cases, it can lead to respiratory failure and even death. The effect of radiation on body tissues is mainly reflected in the direct damage to DNA and the indirect damage to DNA caused by the ionization of water molecules. Cells in the G2 / M phase (G2 / M phase are two key stages in the cell cycle, corresponding to the last preparation before division (G2 phase) and the execution stage of cell division (M phase)) are more sensitive to damage. When radiation acts on lung tissue, it is manifested as damage to alveolar epithelial cells and endothelial cells, causing damage to the alveolar mucosal barrier, thereby triggering a series of inflammatory cascade reactions. Within a few days or weeks, inflammatory cells aggregate, vascular permeability increases, inflammatory cytokines are released, macrophages are activated and aggregated, resulting in local hypoxia, leading to the generation and release of reactive oxygen species and reactive nitrogen species, producing pro-inflammatory, pro-fibrotic, pro-angiogenic and other effects, causing lung tissue damage that persists and forms chronic radiation-induced lung injury. Commonly used drugs for treating radiation-induced lung injury clinically include glucocorticoids and cytotoxic drugs, etc., but long-term use has large side effects and poor efficacy. Therefore, how to effectively treat radiation-induced lung injury has become a major clinical issue. Summary of the Invention
[0003] In view of the deficiencies of the prior art, the present invention provides the application of irisin in the preparation of drugs for treating radiation-induced lung injury.
[0004] The technical solution of the present invention is as follows:
[0005] The application of irisin in the preparation of drugs for treating radiation-induced lung injury.
[0006] Further, the radiation-induced lung injury is caused by X-ray, or γ-ray, or heavy ion radiation.
[0007] Further, the irisin can relieve radiation-induced pulmonary edema.
[0008] Further, the irisin can relieve radiation-induced alveolar interstitial thickening, congestion, alveolar collapse, airway wall thickening, and airway lumen narrowing.
[0009] Further, the irisin can relieve radiation-induced pulmonary fibrosis.
[0010] Furthermore, the irisin can alleviate radiation-induced radioactive pneumonia.
[0011] Furthermore, the radioactive pneumonia can increase the release of IL-β, IL-6, and TNF-α.
[0012] Furthermore, the irisin can alleviate radiation-induced oxidative stress in lung tissue.
[0013] Furthermore, the irisin can alleviate the impairment of the lung tissue barrier function caused by radiation.
[0014] Furthermore, the administration method of the drug includes, but is not limited to, intraperitoneal injection.
[0015] Furthermore, the effective dose of the drug is 1 μg / kg - 10 μg / kg.
[0016] Furthermore, the drug also includes pharmaceutically acceptable excipients, and the excipients include conventional diluents, excipients, fillers, wetting agents, absorption promoters, surfactants, lubricants, or stabilizers in the pharmaceutical field.
[0017] Compared with the prior art, the present invention has at least the following advantages:
[0018] The present invention relates to the application of irisin in the preparation of drugs for treating radiation-induced lung injury. The present invention constructs a C57BL / 6 mouse model of radiation-induced lung injury to explore the effect of irisin on radiation-induced lung injury. The research results prove that exogenous irisin can reduce the thickening of alveolar interstitial tissue, congestion, pulmonary edema, alveolar collapse, airway wall thickening, airway lumen narrowing, and increased peribronchial collagen deposition caused by radiation; in addition, irisin significantly reduces the synthesis and release of inflammatory factors, inflammatory cell infiltration, and oxidative stress in lung tissue caused by radiation, and significantly reduces radiation-induced pulmonary fibrosis and the functional barrier of lung tissue, further demonstrating the role of irisin in resisting radiation-induced lung injury. This not only broadens the research and application of irisin, but also provides new targets and perspectives for the treatment of radiation-induced lung injury. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] In order to more clearly illustrate the specific embodiments of the present invention, the following will briefly introduce the drawings required for the specific embodiments or the description of the prior art.
[0020] Figure 1 This is the result of detecting the degree of pulmonary edema in lung tissue by the wet weight / dry weight method in Example 2 of the present invention;
[0021] Figure 2 This is the result of pathological analysis of lung tissue in Example 3 of the present invention;
[0022] Figure 3Results of the degree of lung tissue fibrosis in Example 4 of the present invention;
[0023] Figure 4 Results of the total protein content in alveolar lavage in Example 5 of the present invention;
[0024] Figure 5 Results of detecting the level of oxidative stress in lung tissue by immunohistochemical staining method in Example 6 of the present invention;
[0025] Figure 6 Results of detecting the content of inflammatory factors in alveolar lavage fluid by ELISA experiment in Example 6 of the present invention;
[0026] Figure 7 Results of the CCK8 viability detection of MLE-12 lung epithelial cells in Example 7 of the present invention. Detailed implementation manners
[0027] The present invention will be further described in detail below. It is necessary to point out here that the following specific implementation manners are only used to further illustrate the present invention and cannot be construed as limiting the protection scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention according to the above application content.
[0028] The present invention generally and / or specifically describes the materials and test methods used in the experiments. The test methods or testing methods involved, unless otherwise specified, are conventional methods; the reagents or instruments used, unless the manufacturer is specified, are commercially available conventional products and are prepared or used by conventional methods.
[0029] Source of materials:
[0030] C57BL / 6 mice: From the Experimental Animal Center of the Third Affiliated Hospital of the Third Military Medical University, 6-8 weeks old, male, weighing 20-23 g;
[0031] Irisin: Irisin recombinant (Human, Rat, Mouse, Canine), purchased from: phoenix pharmaceuticals.
[0032] The following examples use GraphPad Prism statistical analysis software to analyze the experimental data. All data are expressed as "mean ± standard deviation", and ANOVA (one-way analysis of variance) is used for pairwise comparison between groups to analyze the differences. * indicates P < 0.05, with significant statistical significance, and ns indicates P > 0.05, without significant difference; n = 6.
[0033] Example 1 Mouse pretreatment
[0034] C57BL / 6 mice were randomly divided into 3 groups, namely the control group, the irradiation (IR) group, and the irradiation + irisin group (IR + Irisin). The mice in the irradiation (IR) group and the irradiation + irisin group (IR + Irisin) were injected with 120 μl of 1% pentobarbital for anesthesia, fixed in the supine position, and the chest position was marked with white tape. Then they were irradiated with 25 Gy of γ-rays. After the irradiation, the mice in the IR group and the IR + Irisin group were immediately intraperitoneally injected with normal saline and 1 μg / kg irisin respectively, and injected continuously at the same time for 3 - 4 weeks. The body weight of the mice was recorded every day. After 3 - 4 weeks, blood, bronchoalveolar lavage fluid, and lung tissue were collected. Subsequently, the total protein content in the bronchoalveolar lavage fluid was measured and the lung tissue was pathologically analyzed to understand the effect and influence of irisin on radiation-induced lung injury.
[0035] Example 2: Effect of irisin on pulmonary edema
[0036] Using the lung wet weight / dry weight method, first, the lung tissue of C57BL / 6 mice in the experiment was taken and weighed on an electronic balance (wet weight). Then the lung tissue was wrapped in tin foil and placed in an incubator at 60 °C for 48 h, taken out and weighed (dry weight). The wet weight / dry weight (Wet / dry ratio) can be used to evaluate the degree of pulmonary edema in the lung tissue. The results are as Figure 1 shown. It can be seen from Figure 1 the figure that the degree of pulmonary edema in the IR + Irisin group was significantly lower than that in the IR group, indicating that irisin may have the effect of reducing radiation-induced pulmonary edema.
[0037] Example 3: Pathological analysis of lung tissue
[0038] Lung injury score: According to the Smith scoring system, the severity of lung injury was scored separately based on 5 indicators, namely pulmonary edema, infiltration of inflammatory cells in alveoli and interstitium, alveolar and interstitial hemorrhage, atelectasis, and formation of hyaline membranes. If the blood vessels, alveoli, interstitium, and bronchi are all normal, it is 0 point; if the lesion area is less than 25% of the entire field of view, it is 1 point; if the lesion area is 25% - 50% of the entire field of view, it is 2 point; if the lesion area is 50% - 75% of the entire field of view, it is 3 point; if the lesion area is greater than 75% of the entire field of view, it is 4 point; the total score of lung injury is the sum of the above items. The results are as Figure 2 shown. The lung tissue injury in the IR + Irisin group was significantly lower than that in the IR group, indicating that irisin has the effect of reducing radioactive lung injury.
[0039] Example 4: Evaluation of the degree of lung tissue fibrosis
[0040] Analysis was performed using the Masson staining method. The operation was carried out according to the reagent instruction manual, and the fibrotic area was measured by Image J. The results are asFigure 3 As shown, the degree of pulmonary fibrosis in the radiation + irisin group (IR + Irisin) was significantly lower than that in the radiation (IR) group, indicating that irisin may have the effect of reducing radiation-induced pulmonary fibrosis.
[0041] Example 5 Evaluation of the barrier function of lung tissue
[0042] The degree of impairment of the lung tissue barrier function was evaluated by detecting the total protein content (Total protein) in the bronchoalveolar lavage fluid. The results are as Figure 4 shown. The total protein content in the bronchoalveolar lavage fluid of the radiation + irisin group (IR + Irisin) was significantly lower than that in the radiation (IR) group, indicating that irisin can reduce the damage of the lung tissue barrier function caused by radiation.
[0043] Example 6 Irisin reduces radiation-induced inflammatory response and oxidative stress in lung tissue
[0044] Immunohistochemical staining was used to determine oxidative stress in lung tissue. Specifically: Using a rabbit two-step kit (rabbit polymer detection system) (purchased from Zhongshan Jinqiao Biotechnology Co., Ltd.), 4-μm lung tissue sections of experimental C57BL / 6 mice in the control group, radiation (IR) group, and radiation + irisin group (IR + Irisin) were prepared respectively. Then the tissue sections were heated and hydrated at 65 °C, and then dewaxed by washing and soaking in xylene. Subsequently, rehydration was carried out successively with alcohols with volume fractions of 100%, 95%, 80%, and 70%. Then, the tissue sections were treated in a boiling water bath with an immunohistochemical antigen repair buffer (citrate repair solution pH 6.0) for 20 min and rinsed with PBS buffer for 3 min × 3 times; then treated with reagent 1: endogenous peroxidase blocker at room temperature for 20 min and rinsed with PBS buffer for 3 min × 3 times; then blocked with a PBS solution containing 5% goat serum and 1% Triton X-100 at room temperature for 1 h, and 8-OHdG antibody and NOX4 antibody with a dilution concentration of 1:400 were added and incubated overnight at 4 °C. After rinsing with PBS buffer for 5 min × 3 times, reagent 2: enzyme-labeled goat anti-rabbit IgG polymer was added and treated at room temperature for 30 min, rinsed with PBS buffer for 5 min × 3 times, an appropriate amount of freshly prepared DAB chromogenic solution was added and incubated at room temperature for 4 min, rinsed with tap water, and incubated with hematoxylin staining solution (purchased from Beyotime Biotechnology Co., Ltd.) for 5 min, differentiated, rinsed, and blued; finally dehydrated, cleared, and mounted. The results are as Figure 5 shown. As Figure 5 can be seen, compared with the control group, the oxidative stress level measured in the radiation group in the 200× field of view was significantly increased, but it decreased very significantly after using irisin, indicating that irisin has the effect of inhibiting oxidative stress.
[0045] The content of inflammatory factors (IL-1β, IL-6, TNF-α) in bronchoalveolar lavage fluid after radiation was detected by ELISA assay. Specifically, bronchoalveolar lavage fluid was collected from C57BL / 6 mice in the control group, radiation group (IR), and radiation + irisin group (IR + Irisin). Then, the samples were centrifuged at 1500 r for 10 minutes at 4°C, and the supernatant was used to detect the content of IL-1β, IL-6, and TNF-α (the detection kit was purchased from Wuhan Eelite Biotechnology Co., Ltd.). The detection results are as Figure 6 shown. As Figure 6 can be seen, the levels of inflammatory factors in the radiation group were significantly higher than those in the control group, while the use of irisin effectively reduced the levels of oxidative stress indicators and inflammatory factors, indicating that irisin can inhibit inflammatory reactions.
[0046] Example 7 Irisin reduces radiation-induced damage to MLE-12 lung epithelial cells
[0047] The MLE-12 lung epithelial cell line was placed in DMEM solution containing 10% heat-inactivated FBS, 100 U / mL penicillin G, and 100 mg / mL streptomycin by volume. The cultured cells were divided into three groups: the control group without any treatment, the radiation (IR) group, and the radiation + irisin group (IR + Irisin). The MLE-12 cell culture plates in the radiation (IR) group and the radiation + irisin group (IR + Irisin) were treated with 10 Gy of γ-rays, and then 0.1 μg / ml, 0.25 μg / ml, and 0.5 μg / ml irisin were added to the radiation + irisin group (IR + Irisin) immediately, and the samples were collected after 24 hours of treatment.
[0048] The cell viability of the experimental MLE-12 lung epithelial cells in the above three groups was determined by the CCK8 method. The specific method was as follows: 10 μL of CCK8 solution was added to each well of a 96-well cell culture plate and cultured in a cell culture incubator for 1 hour. After the culture, the optical density value of the well plate was detected at a wavelength of 490 nm using an enzyme-linked immunosorbent assay reader. The results are as Figure 7 shown. The results showed that under normal culture conditions, irisin had no obvious effect on cell viability; while in the case of radiation, the use of irisin could restore the OD value decreased by radiation, indicating that irisin could enhance the viability of MLE-12 cells and resist the reduction of cell viability caused by radiation.
[0049] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and the description of the present invention.
Claims
1. Use of irisin in the preparation of a drug for treating radiation-induced lung injury.
2. The application according to claim 1, characterized in that, The radiation-induced lung injury is caused by X-rays, or γ-rays, or heavy ion radiation.
3. The application according to claim 1, characterized in that The irisin can relieve radiation-induced pulmonary edema.
4. The application according to claim 1, wherein The irisin can relieve radiation-induced thickening of alveolar interstitium, congestion, alveolar collapse, thickening of airway wall, and narrowing of airway lumen.
5. The application according to claim 1, wherein The irisin can relieve radiation-induced pulmonary fibrosis.
6. The application according to claim 1, wherein, The irisin can relieve radiation-induced radiation pneumonitis.
7. The application according to claim 1, wherein The irisin can relieve radiation-induced oxidative stress in lung tissue.
8. The application according to claim 1, characterized in that, The irisin can relieve radiation-induced impairment of lung tissue barrier function.
9. The application according to any one of claims 1-8, characterized in that, The administration mode of the drug includes but is not limited to intraperitoneal injection.
10. The application according to claim 9, wherein The effective dose of the drug is 1 μg / kg - 10 μg / kg.