Use of interleukin-41 in preparation of products for preventing and treating allergic rhinitis

Abstract

The application discloses application of interleukin-41 in preparation of products for preventing and treating allergic rhinitis. The interleukin-41 can effectively inhibit the occurrence and development process of allergic rhinitis, effectively alleviate the rhinitis symptoms of mice, improve the inflammation of nasal mucosa and lung tissue, regulate the T cell differentiation in the nasal mucosa and lung tissue of mice, and then effectively regulate the balance of Th cells in the mice, and inhibit the expression of inflammatory cytokines. The application of the interleukin-41 in preparation of products for preventing and treating allergic rhinitis provides a new idea of high efficiency, non-toxicity and safety for preventing and treating allergic rhinitis, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to the application of interleukin-41 (IL-41) in the preparation of products for the prevention and treatment of allergic rhinitis. Background Technology

[0002] Allergic rhinitis (AR), also known as hay fever, is an inflammatory disease of the nasal mucosa mediated by immunoglobulin E (IgE) and caused by allergen exposure. Clinical symptoms of AR mainly include sneezing, runny nose, nasal itching, and nasal congestion, often accompanied by ocular symptoms such as itchy, red, and watery eyes. The main physical signs of AR are pale and swollen nasal mucosa, edema of the inferior turbinate, and copious watery discharge in the nasal cavity. The global prevalence of AR is approximately 5%-50%, and is showing a gradually increasing trend, posing a significant impact on both individuals and society. The personal burden on patients mainly manifests as a decline in quality of life and sleep disturbances, which in turn affect work efficiency and social functioning. The social burden includes increased direct medical costs, increased indirect costs due to lost labor force, and the social costs resulting from a reduced quality of life.

[0003] During the initial stage of a patient's exposure to an allergen without clinical symptoms, dendritic cells in the nasal mucosa process the inhaled allergen. This allergen is then presented to helper T cells (Th0), which activate and differentiate into allergen-specific Th2 cells. Th2 cells secrete cytokines such as interleukin-4 (IL-4) and IL-5, and activate the switching of B cell and IgE classes, leading to B cell differentiation into plasma cells that produce allergen-specific IgE. Once released into the bloodstream, allergen-specific IgE binds to high-affinity IgE receptors on the surface of effector cells such as mast cells and basophils through its unique domain. When the patient is re-exposed to the same allergen, the allergen bridges the allergen with the allergen-specific IgE already bound to the mast cell surface, activating the mast cells and causing them to degranulate, releasing stored and newly synthesized mediators such as histamine, leukotrienes, and prostaglandin D2. These mediators interact with nasal sensory nerves, blood vessels, and glands, leading to the clinical symptoms of acute allergic rhinitis. Simultaneously, patients exhibit inflammatory manifestations such as plasma exudation and type II inflammatory infiltration and mononuclear infiltration characterized by eosinophils, neutrophils, and basophils.

[0004] The daily management of AR patients mainly involves controlling the frequency of exposure to various allergens to reduce the risk of acute AR attacks. AR patients typically receive symptomatic treatment with single or combined medications such as antihistamines, corticosteroids, decongestants, leukotriene receptor antagonists, cromoglycine, and anticholinergics. Interventions at the disease mechanism level, such as biologics and probiotics, can also be used. Allergen immunotherapy (AIT) is one of the main treatment methods for AR, and many studies have affirmed its effectiveness and safety. However, the economic and time costs, as well as various contraindications, make AIT unsuitable for all AR patients. In addition to the above measures, acupuncture, acupoint embedding, and septoplasty can also have a positive impact on disease improvement. In conclusion, the treatment plan for AR patients should be tailored to each patient's specific situation. It is extremely difficult to completely cure AR with a simple, convenient, and universally applicable method. Therefore, there is still an urgent need in this field to find a novel, safe, effective, and clearly defined active substance for the prevention and / or treatment of AR. In view of this, the present invention proposes the application of interleukin-41 in the preparation of products for the prevention and treatment of allergic rhinitis. Summary of the Invention

[0005] The technical problem to be solved by this invention is to provide the application of interleukin-41 in the preparation of products for the prevention and treatment of allergic rhinitis, which addresses the shortcomings of the existing technology and provides a new, efficient, non-toxic, and safe approach to the prevention and treatment of allergic rhinitis.

[0006] The technical solution adopted by the present invention to solve the above-mentioned technical problems is: the application of interleukin-41 in the preparation of products for the prevention and treatment of allergic rhinitis.

[0007] IL-41, also known as Metrnl, is a novel immunomodulatory cytokine or adipokine. IL-41 can be expressed in tissues such as adipose tissue, skin, mucous membranes, liver, and spleen, as well as in activated monocytes and macrophages. Current research has found that IL-41 can act as a neurotrophic factor, nourishing neurotrophic activity, and can also induce white adipose tissue browning, inhibit insulin resistance, and participate in inflammatory responses. Currently, there is limited research on the effects of IL-41 on Th cells. Elevated serum IL-41 levels have been observed in typical Th1-related diseases such as psoriasis and rheumatoid arthritis. In obesity-related diseases, IL-41 can regulate IL-4 expression to affect the disease. In a study of allergic asthma, IL-41 inhibited Th2 cell-related factors such as IL-5 and IL-13 in mice with allergic asthma, alleviating Th2 responses. In septic mice, Chen et al. found that IL-41 deficiency increased serum IL-17 expression, leading to an increase in Th17 cells and a decrease in Treg cells, disrupting the Th17 / Treg balance. The Th1 / Th2 and Th17 / Treg balance system has been extensively studied as one of the pathogenesis mechanisms of acute rhinitis (AR). Although research on the effects of IL-41 on Th cells is currently limited, existing studies suggest its potential to regulate Th cell balance. Research on IL-41 in AR remains largely unexplored, and no related products have emerged. This invention discloses the application of IL-41 in the preparation of products for the prevention and treatment of AR. IL-41 can effectively inhibit the occurrence and progression of AR, effectively alleviate rhinitis symptoms in mice, improve inflammation of the nasal mucosa and lung tissue, effectively regulate the balance of Th cells in mice, and inhibit the expression of inflammatory cytokines, demonstrating broad application prospects.

[0008] In a preferred embodiment, the interleukin-41 is a recombinant interleukin-41 protein.

[0009] Preferably, the dose of the recombinant interleukin-41 protein is 0.05 μg / g body weight.

[0010] In another preferred embodiment, the interleukin-41 is an interleukin-41 plasmid.

[0011] Preferably, the interleukin-41 plasmid is pcDNA3.1-IL-41-His plasmid.

[0012] Preferably, the dose of the interleukin-41 plasmid is 5 μg / g body weight.

[0013] Preferably, the construction process of the pcDNA3.1-IL-41-His plasmid is as follows: using Kpn I and XhoI. The PCR products of pcDNA3.1(+) and interleukin-41 are digested with double enzymes, and the recombinant plasmid formed by linking the digestion products of the two is the pcDNA3.1-IL-41-His plasmid.

[0014] Compared with the prior art, the present invention has the following advantages: 1. Interleukin-41 can effectively inhibit the occurrence and development of allergic rhinitis, effectively alleviate rhinitis symptoms in mice, improve inflammation of nasal mucosa and lung tissue, regulate T cell differentiation in mouse nasal mucosa and lung tissue, and thus effectively regulate the balance of Th cells in mice and inhibit the expression of inflammatory cytokines. 2. The application of interleukin-41 in the preparation of products for the prevention and treatment of allergic rhinitis provides a new, efficient, non-toxic, and safe approach to the prevention and treatment of allergic rhinitis, and has broad application prospects. Attached Figure Description

[0015] Figure 1 The curves showing the changes in body weight of the five groups of mice are shown. Figure 2 Results of nasal symptoms in five groups of mice; Figure 3 HE staining results of nasal mucosa tissue from five groups of mice (×200). Figure 4 HE staining results of lung tissue from five groups of mice (×200); Figure 5 The results of qRT-PCR for T cell-related transcription factors in nasal mucosal tissues from five groups of mice; Figure 6 The results of qRT-PCR of T cell-related effector factors in nasal mucosal tissues from five groups of mice; Figure 7 The results of qRT-PCR for T cell-related transcription factors in lung tissues of five groups of mice; Figure 8 The results of qRT-PCR for T cell-related effector factors in lung tissues of five groups of mice; Figure 9 The results show the detection levels of the inflammatory cytokine IFN-γ in five groups of mice; Figure 10 The results show the detection levels of the inflammatory cytokine IL-4 in five groups of mice; Figure 11 The results show the detection levels of the inflammatory cytokine IL-17 in five groups of mice. Detailed Implementation

[0016] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0017] Construction of pcDNA3.1-IL-41-His plasmid: using Kpn I and Xho I. The PCR products of pcDNA3.1(+) and interleukin-41 are digested with double enzymes, and the recombinant plasmid formed by linking the digestion products of the two is pcDNA3.1-IL-41-His plasmid.

[0018] The following tests were conducted to examine the effects of interleukin-41 on inhibiting the occurrence and development of allergic rhinitis in mice, alleviating inflammation of the nasal mucosa and lung tissue, regulating T cell differentiation in the nasal mucosa and lung tissue of mice, and inhibiting the expression of inflammatory cytokines. I. Interleukin-41 inhibits the occurrence and development of AR in mice SPF-grade male BALB / c mice were randomly divided into five groups: normal saline (NS), ovalbumin (OVA), recombinant mouse IL-41 protein (rmIL-41), pcDNA3.1 empty plasmid (pcDNA3.1), and mIL-41 plasmid (mIL-41). Except for the NS group, the other four groups were used to establish an aeruginosa (AR) model: mice were intraperitoneally injected with 200 μL of OVA aluminum hydroxide suspension on days 0, 7, and 14; and 20 μL of OVA solution was administered intranasally from days 21 to 28. The NS group served as a control with an equal volume of normal saline. The rmIL-41 group received an intraperitoneal injection of 200 μL of normal saline containing 1 μg of recombinant IL-41 protein daily from days 21 to 28. On day 0, the pcDNA3.1 and mIL-41 groups received a tail vein injection of 2 mL containing 100 μg of pcDNA3.1 and mIL-41 plasmids.

[0019] The five groups of mice were fed for a total of 28 days, and their weight was monitored daily. The results of weight changes are shown in the table below. Figure 1 .from Figure 1 It is evident that injection of recombinant IL-41 protein can effectively inhibit weight loss caused by AR disease. Due to the greater stimulation from tail vein injection, the mIL-41 group showed no significant weight gain compared to the empty plasmid group.

[0020] Within 20 minutes after the last nasal stimulation on day 28, the number of sneezes and nasal rubbings in each group of mice were as follows: Figure 2 As shown. Regardless of whether it is in recombinant protein or plasmid form, IL-41 can alleviate nasal symptoms such as sneezing and nose rubbing in mice. This study used OVA to construct an AR mouse model, therefore, the expression of OVA-specific IgE in mouse serum was detected, such as... Figure 2As shown, OVA-IgE expression was significantly increased in the OVA group compared to the NS group, and decreased after treatment with recombinant IL-41 protein and IL-41 plasmid.

[0021] II. Interleukin-41 reduces inflammation of the nasal mucosa and lung tissue. Mice were sacrificed on day 29, and nasal mucosa and lung tissues from the five groups of mice were stained with hematoxylin and eosin (HE). The results are shown in the figures below. Figure 3 and Figure 4 Compared with normal mice, AR mice induced by OVA showed a certain degree of infiltration of inflammatory cells such as eosinophils and lymphocytes in their nasal mucosa and lung tissues, and IL-41 was able to improve the pathological changes in nasal mucosa and lung tissue caused by AR.

[0022] III. Interleukin-41 regulates T cell differentiation in mouse nasal mucosa and lung tissue. The differentiation of T cells in mouse nasal mucosa and lung tissues was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results of T cell-related transcription factors and effector factors in nasal mucosa tissue are shown below. Figure 5 and Figure 6 The results of the lung tissue are shown in Figure 7 and Figure 8 In nasal mucosa tissue, IL-41, whether in recombinant protein or plasmid form, played a regulatory role in the differentiation of Th1, Th2, and Th17 cells in AR mice, but had no significant effect on Treg cells. In lung tissue, although IL-41 protein did not show significant regulation of transcription factor T-bet, effector factors IL-4, and IL-6, a regulatory trend was observed. IL-41 plasmid also showed insignificant regulatory effects on effector factors IL-4 and IL-6. Consistent with nasal mucosa tissue, IL-41 did not have any effect on Treg-type cytokine TGF-β or transcription factor Foxp3 in lung tissue.

[0023] IV. Interleukin-41 inhibits the expression of inflammatory cytokines Serum was collected after euthanizing the mice, and the expression levels of inflammatory cytokines IFN-γ, IL-4, and IL-17 were detected. The results are shown in the table below. Figure 9 , Figure 10 and Figure 11 .Depend on Figures 9-11 It can be seen that IL-41 can inhibit the expression of inflammatory cytokines.

[0024] In summary, interleukin-41 can effectively inhibit the occurrence and development of allergic rhinitis in mice, effectively reduce inflammation of the nasal mucosa and lung tissue, regulate T cell differentiation in the nasal mucosa and lung tissue of mice, and effectively inhibit the expression of inflammatory cytokines.

Claims

1. Application of interleukin-41 in the preparation of products for the prevention and treatment of allergic rhinitis.

2. The application of interleukin-41 according to claim 1 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The interleukin-41 mentioned is a recombinant interleukin-41 protein.

3. The application of interleukin-41 according to claim 2 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The dosage of the recombinant interleukin-41 protein was 0.05 μg / g body weight.

4. The application of interleukin-41 according to claim 1 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The interleukin-41 mentioned is the interleukin-41 plasmid.

5. The application of interleukin-41 according to claim 4 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The interleukin-41 plasmid is pcDNA3.1-IL-41-His plasmid.

6. The application of interleukin-41 according to claim 5 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The dosage of the interleukin-41 plasmid was 5 μg / g body weight.

7. The application of interleukin-41 according to claim 5 in the preparation of products for the prevention and treatment of allergic rhinitis, characterized in that: The construction process of the pcDNA3.1-IL-41-His plasmid is as follows: using Kpn I and Xho I. The PCR products of pcDNA3.1(+) and interleukin-41 are digested with double enzymes, and the recombinant plasmid formed by linking the digestion products of the two is the pcDNA3.1-IL-41-His plasmid.

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