Application of sIL-2R alpha in preparation for early prediction of acute respiratory distress syndrome

Abstract

The invention discloses an application of sIL-2R alpha as a marker in a product for early prediction of acute respiratory distress syndrome. The product for early predicting the acute respiratory distress syndrome comprises a reagent or a medicine for detecting the expression level of sIL-2R alpha in a patient with the acute respiratory distress syndrome. The sIL-2R alpha is used as a biomarker, and experiments prove that the sIL-2R alpha level of the plasma of the ARDS patient is obviously higher than that of a patient without ARDS in the plasma of the ARDS patient. By detecting the sIL-2R alpha level of plasma or serum of an ARDS patient, the ARDS occurrence risk is predicted in an early stage, so that individualized intervention treatment is given, the occurrence rate of ARDS can be reduced, and the kit has important clinical application value for improving prognosis of the ARDS patient.

Description

Technical Field

[0001] The invention belongs to the technical field of biological detection, and specifically relates to an application of sIL-2Rα as a biomarker in a preparation for early prediction of acute respiratory distress syndrome. Background Art

[0002] Acute respiratory distress syndrome (ARDS) is an acute respiratory disease. It is the difficulty and focus of treatment in the field of clinical critical care and is characterized by high morbidity and mortality.

[0003] The pathogenesis of ARDS is very complex, with the characteristics of acute onset and critical condition. Its clinical features are mainly refractory hypoxemia and respiratory distress, and its pathological features are mainly diffuse pulmonary interstitial and alveolar edema caused by damage to pulmonary capillary endothelial cells and alveolar epithelial cells. Early identification and intervention treatment of high-risk patients who have not yet reached ARDS are of great significance to reducing the incidence of ARDS and improving patient prognosis. Although there are many basic research and clinical research projects on ARDS, the intervention measures that can be used in clinical practice for early identification of ARDS and disease progression are still very limited. The main reason is that it is difficult to identify and predict the progression of the disease early, which directly leads to a large number of mild ARDS patients developing moderate to severe ARDS, greatly increasing the duration of ventilator use and the number of days in the ICU, increasing the burden of medical expenses for patients, and seriously threatening patients' lives and prognosis. Early diagnosis is a prerequisite for starting treatment as early as possible. At present, the diagnosis of ARDS is mainly based on etiology, clinical manifestations, oxygenation index, imaging examination, exclusion of cardiogenic pulmonary edema, etc., lacking effective relevant clinical warning factors, which is not conducive to early diagnosis and treatment. Therefore, it is crucial to discover new biomarkers with both high sensitivity and specificity for early prediction of ARDS for its early detection, early diagnosis and early treatment.

[0004] IL-2R is composed of three membrane components, divided into three subunits, including low (α chain), medium (β or, β chain) and high affinity (αβr or αβ) receptors. Different types of IL-2R have different affinities for IL-2. sIL-2Rα is a soluble form of the α chain of IL-2R that is released from the cell membrane into the body fluid. sIL-2Rα is present in large quantities in the blood circulation of healthy people, but it will increase further in patients with infection, inflammation and autoimmune diseases. The main functions of sIL-2Rα in serum are as follows: 1) sIL-2Rα binds to IL-2, protects IL-2 from enzymatic degradation, and prolongs the half-life of IL-2; 2) sIL-2Rα binds to IL-2, increases the affinity of IL-2 to the low-affinity dimer IL-2 receptor; 3) sIL-2Rα binds to IL-2, preventing IL-2 from binding to the low-affinity dimer IL-2 receptor or the high-affinity trimer IL-2 receptor. Studies have shown that low to moderate levels of functional αβγIL-2Rs expressed by pulmonary endothelial cells are the cause of pulmonary edema caused by direct binding of IL-2 to pulmonary endothelial cells, but the specific mechanism has not been explained. Studies have reported that sIL-2Rα can predict the mortality of patients with respiratory failure caused by the new coronavirus, and its continued increase indicates a high inflammatory state, reflecting the severity of the new coronavirus in patients with respiratory failure. In a study of ulcerative colitis (UC), it was confirmed that sIL-2Rα levels reflect the inflammation caused by UC more sensitively than CRP, and it is recommended as a potential biomarker for complete healing of the mucosa in patients with inflammatory bowel disease. The above studies all suggest that sIL-2Rα may become a potential early warning indicator affecting the occurrence and development of the disease and a key target for targeted therapy; however, the research on sIL-2Rα in the early warning of ARDS is still unknown. Summary of the invention

[0005] The present invention aims to solve one of the technical problems in the related art at least to a certain extent. To this end, the main purpose of the present invention is to provide the use of sIL-2Rα in a preparation for early prediction of acute respiratory distress syndrome, aiming to solve the problems of poor sensitivity and specificity of existing biomarkers of acute respiratory distress syndrome.

[0006] The objective of the present invention is achieved through the following technical solutions:

[0007] Application of sIL-2Rα as a marker in products for early prediction of acute respiratory distress syndrome.

[0008] In certain specific embodiments, a product for early prediction of acute respiratory distress syndrome comprises a reagent or a drug for detecting the expression level of sIL-2Rα in patients with acute respiratory distress syndrome.

[0009] In certain specific embodiments, the sIL-2Rα expression level is detected by ELISA double antibody sandwich method to diagnose the product of acute respiratory distress syndrome.

[0010] In certain specific embodiments, a product for early prediction of acute respiratory distress syndrome can predict the occurrence of acute respiratory distress syndrome by detecting the expression level of sIL-2Rα in body fluid samples of patients with acute respiratory distress syndrome. The test results show that the expression level of sIL-2Rα is significantly upregulated compared with normal body fluid samples.

[0011] Furthermore, the body fluid sample is plasma.

[0012] Use of sIL-2Rα in the preparation of products for the prevention or treatment of acute respiratory distress syndrome.

[0013] Description of reagents or drugs:

[0014] Preferably, the agent or drug also includes a pharmaceutically acceptable carrier, such carriers include (but are not limited to): diluents, buffers, suspensions, emulsions, granules, encapsulations, excipients, fillers, adhesives, sprays, transdermal absorbents, wetting agents, disintegrants, absorption enhancers, surfactants, colorants, flavoring agents or adsorption carriers.

[0015] The reagent or drug of the present invention can be prepared into various dosage forms as required, including but not limited to tablets, solutions, granules, patches, ointments, capsules, aerosols or suppositories.

[0016] The administration route of the agent or drug of the present invention is not limited as long as it can exert the desired therapeutic or preventive effect, including but not limited to oral administration, intravenous injection, intramuscular injection, subcutaneous injection, sublingual dissolution, rectal irrigation, nasal spray, oral spray, topical skin administration or systemic transdermal administration.

[0017] The reagent or drug of the present invention can also be used in combination with other drugs for treating acute respiratory distress syndrome. The combined use of multiple drugs can greatly improve the success rate of treatment.

[0018] About the definition content:

[0019] The term "detection" is used herein in the broadest sense to include both qualitative and quantitative measurements of a target molecule. Detection includes merely identifying the presence of a target molecule in a sample as well as determining whether the target molecule is present in the sample at a detectable level.

[0020] As used herein, the term "biomarker" refers to an indicator that can be detected in a sample, such as predictive, diagnostic, and / or prognostic. Biomarkers can serve as indicators of a specific subtype of disease or disorder (e.g., cancer) characterized by certain molecular, pathological, histological, and / or clinical features. In some embodiments, biomarkers are genes. Biomarkers include, but are not limited to, polynucleotides (e.g., DNA and / or RNA), polynucleotide copy number changes (e.g., DNA copy number), polypeptides, polypeptide and polynucleotide modifications (e.g., post-translational modifications), carbohydrates, and / or glycolipid-based molecular markers.

[0021] The term "expression level" refers to the amount of a biomarker in a biological sample. "Expression" generally refers to the process by which information (e.g., genetically encoded and / or epigenetic) is converted into a structure that exists and operates in a cell. Therefore, as used herein, "expression" can refer to transcription into polynucleotides, translation into polypeptides, or even polynucleotide and / or polypeptide modifications (e.g., post-translational modifications of polypeptides). Fragments of transcribed polynucleotides, translated polypeptides, or polynucleotide and / or polypeptide modifications (e.g., post-translational modifications of polypeptides) should also be considered expressed, whether they are derived from transcripts generated by alternative splicing or degraded transcripts, or from post-translational processing of polypeptides, such as by proteolysis.

[0022] The term "diagnosis" is used herein to refer to the identification or classification of a molecular or pathological state, disease or condition.

[0023] As used herein, the term "sample" refers to a composition obtained or derived from a subject and / or individual of interest that contains cells and / or other molecular entities to be characterized and / or identified, for example, based on physical, biochemical, chemical and / or physiological characteristics. For example, the phrase "disease sample" or variations thereof refers to any sample obtained from a subject of interest that is expected to or known to contain cells and / or molecular entities to be characterized. Samples include, but are not limited to, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous humor, lymphatic fluid, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, and tissue culture fluid, tissue extracts such as homogenized tissue, cell extracts, and combinations thereof.

[0024] Compared with the prior art, the present invention has at least the following advantages:

[0025] The present invention uses sIL-2Rα as a biomarker. The applicant has verified through population experiments that the plasma sIL-2Rα level of ARDS patients is significantly higher than that of patients without ARDS. By detecting the plasma or serum sIL-2Rα level, the risk of ARDS can be predicted early, and individualized intervention treatment can be given, which can reduce the incidence of ARDS and has important clinical application value for improving the prognosis of ARDS patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the specific embodiments of the present invention, the drawings required for use in the specific embodiments or the description of the prior art will be briefly introduced below.

[0027] Figure 1 This is a graph showing the results of detecting the expression level of sIL-2Rα in the plasma of ARDS patients and non-ARDS patients in Example 1 of the present invention;

[0028] Figure 2 This is a schematic diagram of the early prediction effect of plasma sIL-2Rα in the risk of ARDS in Example 2 of the present invention using a receiver operating characteristic (ROC) curve;

[0029] Figure 3 This is a graph showing the expression level of sIL-2Rα in the plasma of experimental mice in Example 3 of the present invention. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments, but it should not be understood as limiting the scope of protection of the present invention. Any equivalent changes in form made based on the concept of the present invention should be regarded as the scope of the present invention.

[0031] The present invention provides a general and / or specific description of the materials and test methods used in the experiment. The test methods or test methods involved are all conventional methods unless otherwise specified; the reagents or instruments used are all conventional products available on the market, prepared or used in a conventional manner, unless the manufacturer is specified.

[0032] In the following examples, the research population used was: case data of ARDS patients (183 cases) diagnosed and treated in the PLA Army Special Medical Center from January 2018 to May 2023 were collected, and 50 healthy adult serum specimens were collected as controls. To ensure the consistency of the test results, all were sent to the laboratory of the Army Special Medical Center to detect serum sIL-2Rα levels. All patients participating in the study were approved by the Ethics Committee of the PLA Army Special Medical Center.

[0033] In the following examples, the experimental animals used were: wild-type male C57BL / 6 mice, weighing between 20-22 g and 6-8 weeks old. They were purchased from Chongqing Lepet Biotechnology Co., Ltd. and raised in the SPF-level laboratory of the Experimental Animal Center of the Army Characteristic Medical Center of the Chinese People's Liberation Army. Animal experiments strictly followed the Guidelines for the Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of the People's Republic of China and complied with the management regulations of the Experimental Animal Ethics Committee of the Army Medical University of the Chinese People's Liberation Army. The experimental animal license number was: SCXK(Yu)20170002.

[0034] Example 1

[0035] This example studied the detection of plasma sIL-2Rα levels in ARDS patients and the correlation analysis with the occurrence of ARDS. Specifically:

[0036] By retrospectively collecting the case data of 183 ARDS patients diagnosed and treated at the Army Characteristic Medical Center of the Chinese People's Liberation Army from January 2018 to May 2023, and simultaneously collecting 50 serum specimens of healthy adults (non-ARDS patients) as controls. To ensure the consistency of test results, all were sent to the Clinical Laboratory of the Army Characteristic Medical Center for the detection of serum sIL-2Rα levels. The specific test results are shown in Table 1. All patients participating in the study were implemented after being approved by the Ethics Committee of the Army Characteristic Medical Center of the Chinese People's Liberation Army.

[0037] Table 1 Statistical results of clinical data of ARDS patients

[0038]

[0039]

[0040] The above two groups of detected results (as shown in Table 1) were statistically analyzed using SPSS 22.0 software, and GraphPad Prism 9.0 software was used for drawing. The results are as Figure 1 shown. It can be seen from the figure that compared with non-ARDS patients, the sIL-2Rα level was significantly increased in ARDS patients, and the difference was statistically significant (P<0.0001); indicating that sIL-2Rα can be used as an index for the early prediction of clinical ARDS.

[0041] Example 2

[0042] On the basis of Example 1, this example studied the early prediction value of the ROC curve analysis of plasma sIL-2Rα for the risk of ARDS occurrence. Specifically:

[0043] According to the sIL-2Rα expression level values ​​detected in the plasma of ARDS patients in Example 1, a ROC curve was drawn using SPSS 22.0 software.

[0044] The results are as follows Figure 2 As shown, Figure 2 The figure shows the early prediction effect of plasma sIL-2Rα in the risk of ARDS by receiver operating characteristic curve (ROC). The results show that the area under the curve (AUC) of sIL-2Rα is 0.984, indicating that sIL-2R has a good early prediction value for ARDS.

[0045] Example 3

[0046] This example studies the expression of sIL-2Rα in mouse peripheral blood through animal experiments, specifically:

[0047] Adult C57 mice were selected to establish a sepsis ARDS model. The sepsis ARDS model was established by cecal ligation and puncture (CLP). The specific operation method was as follows: 6-8 weeks old 20-22g C57BL / 6 mice were selected, anesthetized by inhalation of isoflurane, fixed on the operating board, the abdomen was shaved and the skin was disinfected, a 1cm long incision was made in the middle of the abdomen, the colon was ligated, and the cecum at the ligated end was punctured with an 18G syringe needle. After returning to the abdominal cavity, the wound was sutured and the skin was disinfected. The mice were divided into two groups according to sepsis and sham surgery (open and closed abdomen only), and mouse plasma was obtained 24, 48, and 72 hours after modeling for sIL-2Rα protein expression detection.

[0048] The results are as follows Figure 3 As shown, Figure 3 The graph shows the expression level of plasma sIL-2Rα in mice. The results showed that the expression of sIL-2Rα in the plasma of septic ARDS mice was significantly higher than that of the Sham group, especially reaching the highest level 24 hours after modeling, which was also the most serious time for septic ARDS in the CLP model, indicating that sIL-2Rα is likely to be involved in the regulation of the occurrence and development of septic ARDS.

[0049] In summary, the present invention found that the expression level of sIL-2Rα in ARDS patients was significantly higher than that in patients without ARDS. ROC curve analysis showed that sIL-2Rα can be used as a new biomarker for predicting the occurrence of ARDS. Further animal experiments found that sIL-2Rα was expressed at an increased level in the plasma of septic ARDS mice, indicating that sIL-2Rα is likely to be involved in the regulation of the occurrence and development of septic ARDS.

[0050] 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 aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. 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 included in the scope of the claims and specification of the present invention.

Claims

1. Application of sIL-2Rα as a marker in products for early prediction of acute respiratory distress syndrome.

2. The use according to claim 1, characterized in that: Products for early prediction of acute respiratory distress syndrome include reagents or drugs for detecting the expression level of sIL-2Rα in patients with acute respiratory distress syndrome.

3. The use according to claim 1, characterized in that: The product is used to detect the expression level of sIL-2Rα by ELISA double antibody sandwich method to diagnose acute respiratory distress syndrome.

4. The use according to claim 3, characterized in that: A product used for early prediction of acute respiratory distress syndrome. This product can predict the occurrence of acute respiratory distress syndrome by detecting the expression level of sIL-2Rα in body fluid samples of patients with acute respiratory distress syndrome. The test results show that the expression level of sIL-2Rα is significantly upregulated compared with normal body fluid samples.

5. The use according to claim 4, characterized in that: The body fluid sample is plasma.

6. Application of sIL-2Rα in the preparation of products for the prevention or treatment of acute respiratory distress syndrome.

Images