Use of palmitic acid and preparation thereof in preparation of a medicine for preventing and treating acute exacerbation of chronic obstructive pulmonary disease

Abstract

The application relates to the field of biological medicine, and particularly relates to application of palmitic acid and a preparation thereof in preparation of a medicine for preventing and treating acute exacerbation of chronic obstructive pulmonary disease. The application provides application of palmitic acid and the preparation thereof in preparation of the medicine for preventing and treating acute exacerbation of chronic obstructive pulmonary disease. It is found by the application that supplement of palmitic acid in an acute infection period can improve IFITM3 protein expression of alveolar epithelial cells, and further promotes downstream secreted TGFbeta by up-regulating the expression level of an anti-inflammatory cytokine, so that lung tissue inflammation subsides and tissue repair is promoted, and lung function of a patient with chronic obstructive pulmonary disease is improved.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the application of palmitic acid and its preparations in the preparation of drugs for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease. Background Technology

[0002] Chronic obstructive pulmonary disease (COPD) is characterized by persistent respiratory symptoms and irreversible airflow limitation. COPD patients often have congenital and adaptive compromised immune responses. Acute infections in COPD patients are often accompanied by decreased respiratory flora diversity and immune tolerance imbalance. Infection is the primary cause of acute exacerbations of COPD, with viral infections being a significant factor. Studies indicate that COPD patients infected with influenza A virus have an approximately 10% increased risk of respiratory failure.

[0003] Patients with COPD suffer from chronic inflammation, resulting in impaired innate and adaptive immunity, widespread alterations in the state of alveolar epithelial cells, and decreased immune defense function. Consequently, their ability to clear pathogens is significantly reduced when facing infection. COPD patients experience continuous changes in lipid metabolism, with lipid metabolism undergoing reprogramming, affecting the activation and function of immune cells, leading to immune dysregulation and promoting disease progression. Changes in pulmonary lipid metabolism are closely related to decreased lung function, immune dysregulation, and inflammatory responses. The levels of saturated fatty acids such as palmitic acid and stearic acid, and unsaturated fatty acids such as linoleic acid and alpha-linolenic acid in the plasma of COPD patients are significantly lower than in healthy controls. Some studies have shown a positive correlation between fatty acid levels and fatty acid consumption and lung function indicators such as FEV1 and FVC.

[0004] In recent years, in-depth research into the pathophysiological mechanisms of COPD acute exacerbations has revealed that the core involves an abnormally amplified and difficult-to-resolve inflammatory response. Various inflammatory cells (such as neutrophils, macrophages, and T lymphocytes) are activated, releasing large amounts of inflammatory mediators (such as IL-6, IL-8, and TNF-α) and proteases, leading to airway damage, increased oxidative stress, and hypersecretion of mucus. Therefore, developing novel anti-inflammatory drugs targeting this key inflammatory pathway is considered a highly promising strategy for fundamentally preventing and intervening in COPD acute exacerbations. However, existing nonsteroidal anti-inflammatory drugs (NSAIDs) or broad-spectrum immunosuppressants have not been widely used in the prevention and treatment of COPD / AECOPD due to uncertain efficacy or significant systemic toxicity.

[0005] Therefore, there is an urgent need in clinical practice for novel drugs that can effectively regulate COPD-specific inflammatory pathways to fill the current treatment gap in preventing and reducing the occurrence of AECOPD and improve patients' long-term prognosis. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide the application of palmitic acid and its preparations in the preparation of drugs for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease. The present invention provides the use of exogenous palmitic acid to improve lung inflammation during influenza infection in patients with chronic obstructive pulmonary disease, inhibit influenza virus replication, and reduce inflammatory response of alveolar epithelial cells.

[0007] This invention provides the use of palmitic acid and / or its formulations in the preparation of medicaments for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease.

[0008] In some embodiments, the prevention and control includes:

[0009] (1) Downregulates the levels of pro-inflammatory cytokines;

[0010] (2) Upregulates the level of anti-inflammatory cytokines;

[0011] (3) Upregulates the expression of antiviral proteins;

[0012] (4) Improve lung function.

[0013] In some embodiments, the pro-inflammatory factors include at least one of Il1β, Cxcl15, Tnfα, and IFNγ.

[0014] In some embodiments, the anti-inflammatory factors include Socs3, TGFβ, and IL10.

[0015] In some embodiments, the antiviral protein includes IFITM3.

[0016] In some embodiments, the improvement of lung function includes at least one of relieving airflow limitation, prolonging expiratory effort time, reducing respiratory system resistance, reducing airway resistance, increasing inspiratory volume, and increasing static compliance.

[0017] In some embodiments, the acute exacerbation of chronic obstructive pulmonary disease (COPD) is an acute exacerbation of COPD caused by influenza virus.

[0018] In some embodiments, the dosage of palmitic acid is 0.05~0.1 mmol / kg for mice.

[0019] In some embodiments, the formulation comprises palmitic acid liposomes, the palmitic acid liposomes comprising palmitic acid and a phospholipid bilayer encapsulating the palmitic acid;

[0020] The dosage of the palmitic acid liposomes is 0.005~0.01 mmol / kg for mice.

[0021] In some embodiments, the dosage form of the pharmaceutical agent includes tablets, capsules, granules, drops, lyophilized products, granules, ointments, or injections.

[0022] This invention provides the application of palmitic acid and its preparations in the preparation of drugs for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease (COPD). The study of this invention found that palmitic acid supplementation during acute infection can increase the expression of IFITM3 protein in alveolar epithelial cells, and further promote the secretion of TGFβ downstream by upregulating the expression levels of anti-inflammatory cytokines, thereby promoting the resolution of lung inflammation and tissue repair, and ultimately improving lung function in patients with COPD. Attached Figure Description

[0023] Figure 1 This study presents the lung transcriptomics and metabolomics of mice with acute exacerbations of COPD. Figure A shows a volcano plot of differentially expressed genes in the mouse lung tissue transcriptome; Figure B shows a GO enrichment bubble plot of differentially expressed genes in the mouse lung tissue; Figure C shows the gene expression clustering and KEGG enrichment results in the mouse lung tissue transcriptome; Figure D shows a metabolomics enrichment bubble plot in the mouse lung tissue; Figure E shows a volcano plot of differentially expressed fatty acid metabolism-related metabolites in the mouse lung tissue metabolome; and Figure F shows the enrichment results of fatty acid-related metabolic pathways in the mouse lung tissue metabolome. In the figures, NC represents the negative control group, COPD represents the COPD group, and AECOPD represents the acute exacerbation of COPD caused by influenza A virus infection; Up indicates increased expression in the acute relative COPD group, and Down indicates decreased expression in the acute relative COPD group.

[0024] Figure 2 Palmitic acid reduced mortality and lung inflammation levels in mice with acute exacerbation of COPD induced by influenza A virus. Figure A shows the survival curve of COPD mice infected with a lethal dose of influenza A virus 9 days prior; Figure B shows representative lung histopathological images of different groups in the COPD-infected mouse model; Figures C and D show the lung histopathological scores of different groups in the COPD-infected mouse model; and Figures E and H show the Influenza matrix protein in the lungs of different groups in the COPD-infected mouse model. Gene expression levels of nucleoprotein, IL1b, and Cxcl15; Figure I shows the lung TNFα protein level in different groups of a mouse model of COPD infected with influenza A virus; Figure J shows the lung Socs3 gene expression level in different groups of a mouse model of COPD infected with influenza A virus; Figures K and M show the lung TGFβ and IL-10 protein levels in different groups of a mouse model of COPD infected with influenza A virus; NC is marked as the negative control group; COPD is the COPD group; AECOPD is the acute exacerbation group of COPD infected with influenza A virus; AECOPD+PA is the palmitic acid treatment group of acute exacerbation of COPD infected with influenza A virus; *, **** indicate p<0.05; ns indicate p≥0.05;

[0025] Figure 3This study demonstrates how palmitic acid treatment improves lung function in mice with acute exacerbations of COPD. The AE plot shows the levels of lung function indicators FEV0.1, IC, Cst, Rrs, and Rn in different groups of the COPD-H1N1 virus-infected mouse model. The FG plot shows the levels of palmitic acid (PA) and dipalmitoylphosphatidylcholine (DPPC) in different groups of the COPD-H1N1 virus-infected mouse model. NC is marked as the negative control group; COPD represents the COPD group; AECOPD represents the acute exacerbation group of COPD infected with H1N1N1 virus; AECOPD+PA represents the palmitic acid-treated group with acute exacerbation of COPD infected with H1N1N1 virus. *, **** indicate p<0.05; ns indicate p≥0.05.

[0026] Figure 4 The study showed that palmitic acid and palmitic acid-liposomes improved lung inflammation levels and alleviated weight loss in a concentration-dependent manner. Figure A shows representative lung pathological images of mice treated with palmitic acid at doses of 0.025, 0.05, and 0.1 mmol / kg in a mouse model of COPD caused by influenza A virus infection. Figure B shows representative lung pathological images of mice treated with palmitic acid-liposomes at doses of 0.025, 0.05, and 0.1 mmol / kg in the same model. Figure C shows the pathological scores of the lungs of mice treated with palmitic acid-liposomes at doses of 0.025, 0.05, and 0.1 mmol / kg in the same model. Figure D shows the pathological scores of the trachea and blood vessels of mice treated with COPD caused by influenza A virus infection at doses of 0.025, 0.05, and 0.1 mmol / kg in the same model. Figure E shows the pathological scores of the trachea and blood vessels of mice treated with COPD caused by influenza A virus infection. Body weight curves of mice in a mouse model infected with influenza A virus at doses of 0.025, 0.05, and 0.1 mmol / kg over 72 hours. Figure F shows the differences in body weight loss at doses of 0.025, 0.05, and 0.1 mmol / kg over 72 hours in the mouse model of COPD infected with influenza A virus. Figure GH shows the levels of IL-10 and IFNγ proteins in bronchoalveolar lavage fluid at doses of 0.025, 0.05, and 0.1 mmol / kg over 72 hours in the mouse model of COPD infected with influenza A virus. NC is marked as the negative control group; COPD represents the COPD group; AECOPD represents the acute exacerbation group of COPD infected with influenza A virus; AECOPD+PA represents the palmitic acid treatment group of acute exacerbation of COPD infected with influenza A virus; *, **** indicate p<0.05; ns indicate p≥0.05.

[0027] Figure 5Palmitic acid reduces epithelial cell inflammation and promotes IFITM3 protein expression. Figure A shows the UMAP annotation of single-cell GSE268542 cells; Figure B shows the expression levels of pro-inflammatory, anti-inflammatory, and antiviral gene sets in the epithelial cell population of GSE268542; Figure C shows the antiviral gene expression in the epithelial cell population of GSE268542; Figures D and F show the gene expression levels of pro-inflammatory factors Ifnb, Tnf, and Cxcl15 in different groups of mouse lung epithelial cell lines under smoke stimulation; Figure G shows the expression levels of chronic... The expression level of the antiviral protein IFITM3 in a mouse model of chronic obstructive pulmonary disease (COPD) varies with palmitic acid dosage. The H-plot shows the expression level of IFITM3 protein in different groups of mouse lung epithelial cell lines stimulated by smoke. NC is marked as the negative control group. COPD represents the COPD group, AECOPD represents the acute exacerbation group of COPD infected with influenza A virus, and AECOPD+PA represents the palmitic acid-treated group with acute exacerbation of COPD infected with influenza A virus. *, **** indicate p<0.05; ns indicate p≥0.05.

[0028] Figure 6 Palmitic acid promotes the expression of downstream anti-inflammatory proteins in lung epithelial cells. Figure A shows the expression of SOCS3 protein in the lung tissue of mice with acute COPD-H1N1 influenza. Figure B shows the expression level of SOCS3 gene in the lung epithelial cell model. Figure C shows the expression levels of SOCS3 and TGFβ proteins in the lung epithelial cell model. Figure D shows the level of secreted TGFβ in the lung epithelial cell model in culture medium. Figure E shows the protein interaction relationships in the STRING database. Figure F shows the expression levels of STAT3 and p-STAT3 in the lung epithelial cell model. Detailed Implementation

[0029] This invention provides the application of palmitic acid and its preparations in the preparation of drugs for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0030] The inventors discovered through research that exogenous palmitic acid can improve lung inflammation during COPD influenza infection, inhibit influenza virus replication, and reduce inflammatory response of alveolar epithelial cells.

[0031] The purpose of this invention is to promote the sustained expression of the antiviral membrane protein IFITM3 in alveolar epithelial cells by supplementing the lungs with palmitic acid, thereby reducing influenza virus invasion and replication and alleviating epithelial cell inflammation. Simultaneously, it promotes the secretion of the inhibitory cytokines SCOS3 and TGFβ in alveolar epithelial cells, thus promoting inflammation repair.

[0032] The first aspect of the present invention discloses the use of palmitic acid and palmitic acid-liposomes in the treatment of acute influenza virus infection in COPD.

[0033] The second aspect of this invention discloses a drug for treating acute exacerbations of COPD caused by influenza. The drug is a palmitic acid solution and a palmitic acid-liposome solution. In the experiment, the recommended intraperitoneal injection dose for C57BL6 mice (10 weeks old, weighing 25g±2g) is 0.1mmol / kg, and the nebulized inhalation dose is 0.01mmol / kg. During the acute infection period, the drug is administered once daily via intraperitoneal injection or nebulized inhalation.

[0034] The test materials used in this invention are all commercially available products. The invention will be further illustrated below with reference to specific embodiments.

[0035] Example 1: Transcriptomic and metabolomic studies of the lungs in mice with acute exacerbation of COPD

[0036] A mouse model of COPD was established by administering a solution of 350 μg / kg lipopolysaccharide and 60 U / kg elastase in physiological saline via intratracheal infusion in 20 μL doses every 5 days for a total of 6 doses. After COPD modeling, the mice were then infused with 1 × 10⁻⁶ ppm via intratracheal infusion. 7 A PFU / 20 μL influenza virus suspension was used to induce an acute exacerbation model of COPD. Three days after infection, lung tissue and bronchoalveolar lavage fluid from mice were collected for transcriptome sequencing and identification of non-target metabolites.

[0037] The results showed that, Figure 1 The lung transcriptome of COPD mice underwent significant changes after acute influenza virus infection, with multiple infection and inflammation-related pathways significantly upregulated, such as antigen processing and presentation, complement and coagulation cascades, natural killer cell-mediated cytotoxicity, RIG-I-like receptor signaling pathway, cytokine-cytokine receptor interactions, viral protein-cytokine-receptor interactions, and NF-κB signaling pathway. In metabolite identification, lipid metabolism and amino acid metabolism pathways were significantly enriched, including sphingolipid metabolism, fatty acid synthesis, unsaturated fatty acid synthesis, and the tryptophan pathway. Among lipid metabolism-related metabolites, palmitic acid, myristic acid, and arachidonic acid were significantly decreased, while bile acids were significantly increased. Among the pathways enriched for differentially expressed lipid metabolites, fatty acid metabolism, degradation, and synthesis pathways were decreased, while the α-linolenic acid metabolism pathway was upregulated.

[0038] Example 2: Palmitic acid reduces mortality and lung inflammation levels in mice with acute exacerbation of COPD induced by influenza A virus.

[0039] Dissolve 0.325 g of fatty acid-free bovine serum albumin in 8 mL of physiological saline and centrifuge until completely dissolved. This solution will be used to prepare the palmitic acid solution or as a control solvent. Dissolve 6 mg of palmitic acid in 1 mL of 0.1 mol / L sodium hydroxide solution and incubate at 70°C for 30 minutes until clear. Heat the albumin solution to 40°C, and slowly add the clear palmitic acid solution to the albumin solution. Gently shake until clear, adjust the pH to 7.4, filter through a 0.4 μm sterile filter, and dispense into 5 mM palmitic acid solutions. The therapeutic dose is 0.1 mmol / kg, administered via intraperitoneal injection.

[0040] C57BL / 6N mice were randomly divided into four groups, with six mice in each group. The control group consisted of mice that received no treatment. The COPD group consisted of mice that underwent endotracheal infusion of 50 μg / kg lipopolysaccharide and 60 U / kg elastase six times. The COPD acute exacerbation group consisted of mice that underwent COPD modeling after 5 × 10⁻⁶ treatments. 7 Mice infected with PFU influenza virus via intratracheal instillation. The COPD acute exacerbation treatment group consisted of mice treated with palmitic acid solution via intraperitoneal injection during the acute infection period to establish a COPD model, from the day of infection to day 4 post-infection. Survival experiment mice were infected with 5 × 10⁶ PFU / mL. 7 The mice were treated with PFU / 20μL of influenza virus until the end of the observation period.

[0041] Mouse lung tissue was collected, and total RNA was extracted for reverse transcription. cDNA was diluted to prepare the reaction mixture, and the amplification program was 60℃ for 30 seconds, 95℃ for 10 seconds, for 40 cycles. Data analysis was performed using relative quantification.

[0042] Mouse lung tissue was immersed in tissue fixative for 24 hours, followed by graded ethanol dehydration, xylene clearing, and paraffin embedding. Paraffin sections were sectioned, mounted on glass slides, and baked at 60°C for 30 minutes. After dewaxing, hematoxylin and eosin staining were performed, and the slides were mounted with neutral resin for observation of lung tissue structure, septal thickness, and inflammatory infiltration.

[0043] The results showed that, Figure 2In mice with COPD infected with influenza virus, mortality began on day 4, reaching 75% by day 9, with no difference between the solvent control group and the untreated group. Mice receiving intraperitoneal injection of palmitic acid began dying on day 6, and by day 9, the mortality rate was significantly lower than that of the untreated and solvent control groups, at 50%, indicating that palmitic acid supplementation delayed the time to death and reduced mortality in COPD mice infected with influenza. Pathological examination of mouse lungs showed that after 4 days of palmitic acid treatment, lung inflammation in mice with acute COPD infection significantly improved. Hematoxylin and eosin staining showed a significant reduction in inflammatory infiltration, and the inflammation scores of the bronchi, blood vessels, and alveoli were lower in the treated group compared to the untreated group. RT-qPCR showed that, compared to the untreated group, the transcription levels of influenza virus M and N proteins in the lung tissue of mice in the treated group decreased, the levels of Il1β, Cxcl15, and Tnfα decreased, while the level of Socs3 increased. ELISA analysis of mouse bronchoalveolar lavage fluid showed that the levels of anti-inflammatory proteins TGFβ and IL10 in the mouse lungs increased after palmitic acid treatment.

[0044] Example 3: Palmitic acid treatment improves lung function in mice with acute exacerbations of COPD.

[0045] Mice in the four groups were anesthetized by intraperitoneal injection of sodium pentobarbital. After their vital signs stabilized, tracheostomies were performed. The small animal pulmonary function instrument was then calibrated, and the mice were placed in a supine position and connected to the instrument to perform two deep breaths. Lung function parameters were measured.

[0046] The results are as follows Figure 3 As shown, palmitic acid treatment during influenza virus infection in COPD mice alleviated airflow limitation to some extent, prolonged forced expiratory time (FEV1), decreased respiratory and airway resistance, and partially restored inspiratory volume (IC) and static compliance (Cst). Simultaneously, the levels of palmitic acid and dipalmitoylphosphatidylcholine in the bronchoalveolar lavage fluid increased, suggesting that intraperitoneal injection of palmitic acid can effectively replenish the palmitic acid content in the lungs, increase the synthesis of alveolar surfactant, and thus promote the recovery of lung function during acute exacerbations of COPD.

[0047] Example 4: Palmitic acid and palmitic acid-liposomes improve lung inflammation levels and alleviate weight loss in a concentration-dependent manner.

[0048] Lecithin, cholesterol, and palmitic acid were dissolved in chloroform and methanol (2:1), and the mixture was rotary evaporated at 40°C to form a homogeneous film, which was then vacuum dried for 1 hour. The film was hydrated with preheated phosphate-buffered saline, sonicated in a water bath for 15 minutes, and then centrifuged to remove free palmitic acid and impurities, yielding palmitic acid-liposomes. The therapeutic doses were 0.0025 mmol / kg, 0.005 mmol / kg, and 0.01 mmol / kg, administered via intratracheal infusion.

[0049] Mice were randomly divided into a blank control group, a COPD group, a COPD acute exacerbation group, and a treatment group with palmitic acid doses of 0.025 mmol / kg, 0.05 mmol / kg, and 0.1 mmol / kg, respectively, with 3 mice in each group. Body weight changes were recorded over 3 days. In the palmitic acid-liposome treatment experiment, mice were divided into a blank control group, a COPD group, a COPD acute exacerbation group, and a treatment group with palmitic acid-liposome doses of 0.0025 mmol / kg, 0.005 mmol / kg, and 0.01 mmol / kg, with 3 mice in each group. Administration continued until day 4 post-acute infection. Four days later, mice were euthanized, and bronchoalveolar lavage fluid and lung tissue were collected.

[0050] The results showed that, Figure 4 In mice, lung inflammation was reduced with increasing palmitic acid dosage, with the best reduction observed at intraperitoneal injection (0.1 mmol / kg) and intratracheal infusion (0.01 mmol / kg). At an intraperitoneal injection of 0.1 mmol / kg, alveolar, bronchial, and small vessel inflammation was significantly reduced, with minimal weight loss. Furthermore, the levels of the anti-inflammatory factor IL10 in bronchoalveolar lavage fluid were increased, while IFNγ levels were significantly decreased, indicating that palmitic acid promoted the resolution of inflammation in mice during acute exacerbations.

[0051] Example 5: Palmitic acid reduces lung epithelial cell inflammation and promotes IFITM3 protein expression.

[0052] Transfer 10 mL of serum-free culture medium to a 50 mL centrifuge tube. Take two sterile glass tubes of appropriate length, attaching one end below the surface of the culture medium and the other above. Connect the longer tube to a lit cigarette and the shorter tube to a negative pressure aspirator. Slowly burn 10 cigarettes to allow the smoke to fully dissolve in the serum-free culture medium. Pass the solution through a 0.22 μm sterile filter to obtain a 100% concentration of cigarette smoke extract (CSE). Use 0.5% of this extract for cell experiments.

[0053] Cellular experiments were performed using the mouse alveolar epithelial cell line MLE12. Cells in good condition were collected and divided into four groups: a blank control, smoke-exposed cells, smoke-exposed cells combined with influenza virus infection, and a smoke-exposed cell combined with influenza virus infection treatment group. Cells in the smoke-exposed cell combined with influenza virus infection were infected with influenza virus one day after the addition of CSE, and the medium was replaced with CSE-containing medium two hours later. In the treatment group, 150 μM palmitic acid was added simultaneously with infection, and 150 μM palmitic acid was added again after medium replacement. Cells were harvested one day later, and cellular RNA was extracted for real-time quantitative polymerase chain reaction (qPCR). Cells were also harvested, and proteins were extracted for Western blotting analysis.

[0054] Mouse lung tissue was embedded in paraffin and dewaxed with xylene, then rehydrated with graded ethanol. Citrate buffer was used for heat retrieval according to the antibody instructions, followed by natural cooling and rinsing with PBS. After blocking, the tissue was incubated with primary and secondary antibodies sequentially. DAB staining was performed to the desired intensity, followed by rinsing, counterstaining with hematoxylin, bluing, dehydration, clearing, and then mounting with resin. Staining intensity was quantified using ImageJ software.

[0055] The results are as follows Figure 5 As shown, epithelial cells isolated from single-cell data (GSE268542) of lung tissue from COPD patients 24 hours after influenza virus infection were found to have enhanced pro-inflammatory responses in epithelial cells after infection, but decreased levels of anti-inflammatory proteins, and decreased expression of some antiviral proteins. In MLE12 cells, the addition of palmitic acid significantly reduced cellular inflammation levels and increased the expression of the antiviral protein IFITM3. In animal studies, the expression of the antiviral protein IFITM3 also increased with increasing palmitic acid dosage.

[0056] Example 6: Palmitic acid promotes the expression of downstream anti-inflammatory proteins in lung epithelial cells.

[0057] Cell experiments were conducted using a 0.5% concentration of smoke-induced irritant extract and the mouse alveolar epithelial cell line MLE12. Cells in good condition were divided into a blank control, a smoke-induced irritant group, a smoke-induced irritant combined with influenza virus infection, and a smoke-induced irritant combined with infection treatment group. Cells in the smoke-induced irritant combined with influenza virus infection were infected with influenza virus one day after the addition of CSE, and the medium was replaced with CSE-containing medium two hours later. In the treatment group, 150 μM palmitic acid was added simultaneously with infection, and 150 μM palmitic acid was added again after medium replacement. Cells were harvested one day later, and cellular RNA was extracted for real-time quantitative polymerase chain reaction (qPCR). Cells were also harvested, and proteins were extracted for Western blotting analysis. Protein-protein interaction relationships were retrieved using the STRING protein interaction database.

[0058] The results are as follows Figure 6 As shown, palmitic acid treatment enhanced the expression of the anti-inflammatory protein SOCS3 in the lungs of mice infected with influenza virus in response to smoke stimulation. In the cell model, the expression of anti-inflammatory proteins SOCS3 and TGFβ was partially restored, and the promoting effect of IFITM3 on downstream anti-inflammatory proteins may be mediated by phosphorylation of STAT3 protein.

[0059] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of palmitic acid and / or its preparations in the preparation of drugs for the prevention and treatment of acute exacerbations of chronic obstructive pulmonary disease.

2. The application according to claim 1, characterized in that, The prevention and control measures include: (1) Downregulates the levels of pro-inflammatory cytokines; (2) Upregulates the level of anti-inflammatory cytokines; (3) Upregulates the expression of antiviral proteins; (4) Improve lung function.

3. The application according to claim 2, characterized in that, The pro-inflammatory factors include at least one of Il1β, Cxcl15, Tnfα, and IFNγ.

4. The application according to claim 2, characterized in that, The anti-inflammatory factors include Socs3, TGFβ, and IL10.

5. The application according to claim 2, characterized in that, The antiviral protein includes IFITM3.

6. The application according to claim 2, characterized in that, The improvement in lung function includes at least one of relieving airflow limitation, prolonging expiratory effort time, reducing respiratory system resistance, reducing airway resistance, increasing inspiratory volume, and increasing static compliance.

7. The application according to any one of claims 1 to 6, wherein the acute exacerbation of chronic obstructive pulmonary disease is an acute exacerbation of chronic obstructive pulmonary disease caused by influenza virus.

8. The application according to any one of claims 1 to 7, characterized in that, The dosage of palmitic acid is 0.05~0.1 mmol / kg for mice.

9. The application according to any one of claims 1 to 7, characterized in that, The formulation includes palmitic acid liposomes, which include palmitic acid and a phospholipid bilayer encapsulating the palmitic acid. The dosage of the palmitic acid liposomes is 0.005~0.01 mmol / kg for mice.

10. The application according to any one of claims 1 to 9, characterized in that, The dosage forms of the medicine include tablets, capsules, granules, drops, lyophilized products, granules, ointments, or injections.

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