Application of ILF3 gene in diagnosis or treatment of sepsis

Abstract

The invention relates to application of an ILF3 gene in diagnosis or treatment of sepsis, and belongs to the technical field of biological medicines. It is found for the first time that the ILF3 gene is a macrophage-related differential expression gene of sepsis, the expression level of the ILF3 gene in plasma and alveolar lavage fluid of sepsis patients is obviously higher than that of healthy people, a primer pair capable of specifically recognizing and detecting the ILF3 gene is designed, a mouse with the ILF3 gene in macrophage being specifically knocked out is constructed, and the ILF3 gene in the macrophage is specifically knocked out. The invention also designs si-ILF3 capable of specifically interfering with the expression level of the ILF3 gene. The ILF3 gene is used as a sepsis diagnosis marker and a sepsis treatment target, the application of the ILF3 gene detection reagent in preparation of sepsis diagnosis products and the application of si-ILF3 in preparation of sepsis treatment drugs are provided, and the si-ILF3 gene detection reagent is used for further diagnosis and treatment of sepsis patients.

Description

Technical Field

[0001] The present invention relates to application of ILF3 gene in diagnosing or treating sepsis, and belongs to the technical field of biomedicine. Background Art

[0002] Sepsis is a serious life-threatening disease characterized by organ dysfunction caused by an abnormally regulated host response to infecting pathogens. In 2020, sepsis was defined by the World Health Organization as a major global health problem. The lungs, urinary tract, and abdominal cavity are the preferred sites of sepsis-related infection. More than 50% of sepsis patients will develop acute lung injury (ALI) and its progressive form, acute respiratory distress syndrome (ARDS), which is characterized by destruction of the vascular endothelium and alveolar epithelium, increased alveolar capillary permeability, intrapulmonary hemorrhage, and decreased lung function. In the lungs, the over-activated inflammatory response triggered by massive infiltration of immune cells is closely related to the pathogenesis of ALI / ARDS.

[0003] However, the pathogenesis of sepsis is highly complex, involving a series of physiological and pathological processes, including infection, inflammation, immunity, coagulation, and tissue damage. The inflammatory factors released by infection mobilize multiple cell and organ systems throughout the body through the neuro-endocrine-immune system, forming a complex network of cascade amplification and mutual restraint. Therefore, clinical treatment is extremely difficult. Treatment methods mainly rely on antimicrobial therapy and comprehensive treatment targeting alterations in various systems. Currently, there is a lack of effective medical methods to increase bacterial clearance rates in sepsis and improve the survival rate of septic patients.

[0004] Interleukin enhancer binding factor 3 (ILF3) encodes a double-stranded RNA (dsRNA) binding protein consisting of two major isoforms: NF90 and NF110. It is involved in diverse physiological activities, including mRNA transport, translation, transcription, and signal transduction. Previous studies have identified ILF3 as a novel factor influencing dyslipidemia and stroke subtypes. Recent studies have demonstrated that ILF3 plays a key role in regulating inflammatory responses. MiR-215-5p suppresses the inflammatory response in H9c2 cells by regulating ILF3. Furthermore, macrophage-derived ILF3 accelerates abdominal aortic aneurysm by inducing inflammatory imbalance. However, research on the role of ILF3 in the diagnosis and treatment of sepsis remains scarce. Summary of the Invention

[0005] In view of the deficiencies of the existing technology, the present invention provides the use of the ILF3 gene in the diagnosis or treatment of sepsis.

[0006] The technical solutions of the present invention are as follows:

[0007] Application of a reagent for detecting ILF3 gene expression levels in the preparation of sepsis diagnostic products.

[0008] Preferably according to the present invention, the nucleotide sequence of the ILF3 gene is shown as SEQ ID NO.1.

[0009] Preferably, according to the present invention, the reagent for detecting the expression level of human ILF3 gene comprises the primer pair shown as SEQ ID NO. 2-3.

[0010] Preferably, according to the present invention, the detection sample of the reagent for detecting the expression level of human ILF3 gene is selected from cells, tissues, plasma or serum.

[0011] Application of ILF3 gene as a drug target in the preparation of drugs for the treatment of sepsis.

[0012] Preferably according to the present invention, the nucleotide sequence of the ILF3 gene is shown as SEQ ID NO.1.

[0013] Preferably, according to the present invention, the drug for treating sepsis uses the ILF3 gene as an intervention target and can efficiently and specifically downregulate the expression level of the ILF3 gene.

[0014] Application of siRNA that specifically interferes with the expression level of ILF3 gene in the preparation of drugs for treating sepsis.

[0015] Preferably, according to the present invention, the nucleotide sequence of the siRNA that specifically interferes with the expression level of the ILF3 gene is as shown in SEQ ID NO. 4 and 5.

[0016] A drug for treating sepsis, comprising siRNA that specifically interferes with the expression level of the ILF3 gene.

[0017] Preferably, according to the present invention, the nucleotide sequence of the siRNA that specifically interferes with the expression level of the ILF3 gene is shown as SEQ ID NO.4.

[0018] Beneficial effects:

[0019] The present invention discovered for the first time that the ILF3 gene is a differentially expressed gene associated with macrophages in sepsis. Its expression levels in the plasma and bronchoalveolar lavage fluid of sepsis patients are significantly higher than those in ICU patients and pneumonia patients. A primer pair capable of specifically identifying and detecting the ILF3 gene was designed, mice with specific knockout of the ILF3 gene in macrophages were constructed, and si-ILF3, a specific inhibitor of ILF3 gene expression, was designed. Experiments then verified that mice with specific knockout of the ILF3 gene in macrophages showed significantly reduced sepsis symptoms, significantly improved lung tissue apoptosis, and significantly suppressed inflammatory cell infiltration and inflammatory response. Therefore, the present invention uses the ILF3 gene as a diagnostic marker for sepsis and a target for sepsis treatment. It proposes the use of ILF3 gene detection reagents in the preparation of sepsis diagnostic products and the use of si-ILF3 in the preparation of drugs for treating sepsis, for further diagnosis and treatment of sepsis patients, while also participating in the study of new mechanisms of sepsis progression. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 To detect the expression level of ILF3 in plasma samples from ICU patients and sepsis patients and in bronchoalveolar lavage fluid samples from pneumonia patients and sepsis patients;

[0021] In the figure, A is the expression level of ILF3 protein in plasma samples of ICU patient controls and sepsis patients; B is the expression level of ILF3 protein in bronchoalveolar lavage fluid samples of pneumonia patients and sepsis patients; C is the expression level of ILF3 mRNA in plasma samples of ICU patient controls and sepsis patients; D is the expression level of ILF3 mRNA in bronchoalveolar lavage fluid of pneumonia patients and sepsis patients.

[0022] Figure 2 The receiver operating characteristic curve (ROC) was used to evaluate the specificity and sensitivity of ILF3 gene expression levels in the diagnosis of sepsis.

[0023] Figure 3 Schematic diagram of the process of constructing macrophage ILF3 conditional knockout mice and the results of ILF3 gene knockout;

[0024] In the figure, A is a schematic diagram of the process; B is the ILF3 gene knockout efficiency detection result.

[0025] Figure 4 The survival rates of mice in different groups at 0, 12, 24, 36, and 48 hours.

[0026] Figure 5 The lung tissue damage results of mice in different groups;

[0027] In the figure, A is HE staining; B is lung dry / wet weight ratio; C is Evans blue staining; D is Occludin fluorescence staining.

[0028] Figure 6 The results of cell apoptosis in different groups of mice;

[0029] In the figure, A is Tunel staining; BE are Western Blot detection of Bax, Bcl-2, and Cleaved caspase-3 expression levels.

[0030] Figure 7 The changes in inflammation levels of mice in different groups;

[0031] In the figure, A is the immunohistochemical staining of the macrophage marker CD68 and the inflammatory markers IL-6, TNF-α, and MCP-1; B is the changes in the levels of IL-1β, IL-6, IL-18, and TNF-α in the alveolar lavage fluid; C is the changes in the levels of IL-1β, IL-6, IL-18, and TNF-α in the plasma.

[0032] Figure 8 This is the knockout efficiency test result of si-ILF3 knockout of ILF3 gene;

[0033] In the figure, A is the detection of knockout efficiency at the protein level; B is the detection of knockout efficiency at the RNA level. DETAILED DESCRIPTION

[0034] To make the purpose, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention are described below. It should be pointed out that the described embodiments are part of the present invention, and all other embodiments obtained by other technicians in this field based on the embodiments of the present invention without making any creative efforts are within the scope of protection of the present invention.

[0035] Unless otherwise specified, the raw materials, reagents, or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0036] Example 1

[0037] 1. Collect samples

[0038] Blood samples were collected from 23 patients with sepsis (Sepsis) and 27 ICU patients (Control). Whole blood was collected using EDTA or sodium citrate anticoagulant tubes to prevent heparin interference in subsequent experiments. The blood was then centrifuged at 1500g for 10 minutes at 4°C, and the upper plasma layer was carefully aspirated. Albumin and IgG were removed using a specialized kit. CaCl2 (final concentration 10 mM) was added to the plasma to promote fibrin coagulation. After centrifugation, the supernatant was collected to obtain plasma samples (human peripheral blood mononuclear cells).

[0039] Bronchoalveolar lavage fluid (BALF) was collected from 37 patients with acute lung injury due to sepsis (Sepsis) and 30 patients with pneumonia (Control). The fluid was immediately placed on ice and centrifuged at 500 g for 10 minutes at 4°C to remove cellular debris. The supernatant was transferred to a fresh tube. The fluid was concentrated to 100-200 μL using a 10 kDa ultrafiltration tube and centrifuged at 4000 g for 30 minutes at 4°C. An equal volume of PBS buffer was added for dilution, the fluid was vortexed, and the mucus precipitate was removed by centrifugation at 10000 g for 10 minutes at 4°C to obtain human BALF samples.

[0040] Among them, ICU patients are those who are not infected but have other systemic diseases.

[0041] 2. Detection of ILF3 protein expression levels in plasma samples and bronchoalveolar lavage fluid samples

[0042] Total protein in plasma samples and bronchoalveolar lavage fluid samples was extracted using RIPA lysis buffer containing 1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS. The expression levels of ILF3 protein in plasma samples and bronchoalveolar lavage fluid samples were then detected by Western blot. The results are shown in Figure 3. Figure 1 As shown in A and B.

[0043] The specific method is as follows:

[0044] Sodium dodecyl sulfate polyacrylamide (SDS-PAGE) gels were prepared with 10% separating gel and 5% stacking gel, with 25 μg of protein loaded and electrophoresis performed at a constant voltage of 90 V for the stacking gel and 120 V for the separating gel. Subsequently, the membrane was transferred at a constant current of 200 mA for 2 h and blocked with 5% skim milk powder at room temperature for 2 h. After blocking, the membrane was washed with TBST buffer and incubated in primary antibodies (ILF3 (Abcam, ab92355) and β-actin (Abcam, ab8245)) at 4°C overnight. After the primary antibody incubation, the membrane was washed with TBST buffer and incubated in secondary antibodies (Zhongshan Jinqiao, ZB5301 and ZB2305) at room temperature for 1 h. After the secondary antibody incubation, the membrane was washed with TBST buffer, ECL luminescent solution (Millipore, WBKLS0500) was added dropwise, and the membrane was exposed in a luminometer.

[0045] Depend on Figure 1 As shown in Figures A and B, compared with ICU patients, the expression level of ILF3 protein in peripheral blood mononuclear cells of sepsis patients was significantly increased; compared with pneumonia patients, the expression level of ILF3 protein in bronchoalveolar lavage fluid of sepsis patients was significantly increased.

[0046] 3. Detection of ILF3 gene expression levels in plasma samples and bronchoalveolar lavage fluid samples

[0047] Total RNA was extracted from plasma and bronchoalveolar lavage fluid supernatants according to the Quiamp MinElute Virus Vacuum Kit procedure. The expression levels of ILF3 genes in plasma and bronchoalveolar lavage fluid samples were then detected by q-PCR. The results are shown in Figure 2. Figure 1 As shown in C and D.

[0048] The specific method is as follows: total RNA from plasma and bronchoalveolar lavage fluid supernatant was extracted using Quiamp MinElute Virus Vacuum Kit and reverse transcribed to obtain cDNA; then, q-PCR amplification was performed using cDNA as template using TB Green and primers shown in SEQ ID NO. 2-3, with GAPDH expression level as internal reference and ILF3 relative expression level as 2 -ΔΔCT Method calculation.

[0049] Among them, the sequence shown in SEQ ID NO.2 is: 5'-CATTACGCCCATGAAACGCC-3',

[0050] The sequence shown in SEQ ID NO. 3 is: 5'-TAAAGATGGGGGCATGGACG-3'.

[0051] Depend on Figure 1As shown in Figures C and D, the expression level of ILF3 gene in plasma of patients with sepsis was significantly increased compared with that in ICU patients; and the expression level of ILF3 gene in bronchoalveolar lavage fluid of patients with sepsis was significantly increased compared with that in patients with pneumonia.

[0052] In summary, by extracting plasma samples from ICU patients and sepsis patients, and bronchoalveolar lavage fluid samples from pneumonia patients and sepsis patients, extracting proteins and RNA for Western blot and PCR experiments, it was confirmed that the level of ILF3 in sepsis patients was significantly increased, indicating that ILF3 has a significant effect on sepsis.

[0053] Example 2

[0054] The expression level data of ILF3 gene in the plasma samples tested in Example 1 were collected, and the receiver operating characteristic curve (ROC) was used to evaluate the specificity and sensitivity of ILF3 gene expression level in the diagnosis of sepsis. The test results are as follows: Figure 2 shown.

[0055] Depend on Figure 2 The area under the ROC curve (AUC) was 0.847 (P < 0.0001, CI: 0.788-0.894). When the Youden index was at its maximum, its specificity was 80.65% and its sensitivity was 77.91%. This indicates that ILF3 gene expression levels have good efficacy for the clinical diagnosis of sepsis and can be used to develop products such as sepsis diagnostic kits.

[0056] Example 3

[0057] 1. Construction of ILF3 conditional knockout mice

[0058] like Figure 3 As shown in A, 8-week-old male mice were selected and the DNA of the intron in the ILF3 gene was cut using CRISPR technology. At the same time, a homologous template Donor was provided and FloxP was inserted at both ends of the specific exon of the mouse ILF3 gene through DNA homologous recombination repair technology to obtain ILF3 flox / flox Then ILF3 flox / flox Mice are mated with macrophage-specific Cre recombinase-expressing mice (LysMCre), which results in the deletion of specific exons of the ILF3 gene. The ILF3 gene will not be translated or a frameshift mutation will occur, leading to the inactivation of the ILF3 protein. This achieves conditional knockout of the ILF3 gene in macrophages. This mouse is a mouse that specifically knocks out the ILF3 gene in macrophages. M-KO express.

[0059] Then extract ILF3 flox / flox Mice and ILF3 M-KO The expression level of ILF3 protein was detected by Western blot in the bone marrow macrophages of mice according to the method described in Example 1. Figure 3 As shown in B.

[0060] Depend on Figure 3 As shown in Figure B, ILF3 levels in macrophages of ILF3 gene-specific knockout mice were significantly reduced, indicating that the successful specific knockout of the ILF3 gene in macrophages provides a reliable guarantee for subsequent experiments.

[0061] 2. Construction of sepsis model mice

[0062] ILF3 flox / flox Mice and ILF3 M-KO The mice were randomly divided into 4 groups, 10 in each group, and were designated as ILF3 flox / flox Group, ILF3 M-KO group, sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO Group.

[0063] The above four groups of mice were treated differently, as follows:

[0064] ILF3 flox / flox Group and ILF3 M-KO Mice in each group were placed in an environment with a 12-h dark / light cycle, a temperature of 20-26°C, and a humidity of 40-50%, with free access to food and water.

[0065] Sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO The sepsis model mice were established according to the following method.

[0066] Construction Method: Under sterile conditions, the mouse abdominal skin was incised, the cecum precisely located, and 75% of it ligated with 6-0 silk suture. The ligated portion was then perforated and the contents squeezed out. Finally, the cecum was reinserted into the abdominal cavity and the skin sutured. For sham-operated mice, the cecum was not ligated or punctured; all other procedures were the same.

[0067] Among them, sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO Mice in the two groups were intraperitoneally injected with 250 μg of anti-IgG or anti-mouse Ly6G antibodies 24 hours before modeling to eliminate neutrophils.

[0068] 3. Comparison of survival rates of mice in different groups

[0069] In Sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO After the model was successfully established in the mice group, ILF3 flox / flox 、ILF3 M-KO 、sepsis+ILF3 flox / flox and sepsis+ILF3 M-KO The survival of these four groups of mice at 0, 12, 24, 36, and 48 hours, and the survival rates were calculated. The results are as follows Figure 4 shown.

[0070] Depend on Figure 4 It can be seen that ILF3 flox / flox 、ILF3 M-KO Mice survived normally. flox / flox Compared with the mice in the sepsis+ILF3 group, the ILF3 gene in macrophages was specifically knocked out. M-KO The survival rate of mice in the control group was significantly improved, indicating that knocking out the ILF3 gene and inhibiting its expression can effectively treat sepsis and reduce the mortality rate of sepsis.

[0071] 4. Comparison of lung tissue damage in different groups of mice

[0072] Remove ILF3 flox / flox Group, ILF3 M-KO group, sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO The lung tissues of the mice in the 2 groups were then paraffin-embedded and sectioned, and HE staining, Occludin fluorescence staining, and lung dry / wet weight ratio were measured. The results are as follows: Figure 5 As shown in A, C, and D. At the same time, Evans blue staining was performed on the surviving mice to evaluate the pulmonary vascular permeability of the mice. Figure 5 As shown in B.

[0073] HE staining and Occludin fluorescence staining are both conventional existing techniques and can be performed according to existing methods.

[0074] The lung dry / wet weight ratio was measured by isolating the mouse lung tissue and absorbing the surface liquid with clean gauze and measuring the wet weight. The tissue block was then dried in a 60°C oven for 48 hours, and the dry weight was measured. Finally, the lung tissue dry / wet weight ratio was calculated based on the mass change.

[0075] Evans blue staining method: After anesthetizing the mice, 0.5% Evans blue solution (2 ml / kg) was injected through the tail vein; 1 hour later, the mice were anesthetized again and perfused with normal saline to remove the dye in the blood vessels. Subsequently, part of the lung tissue (100 mg) was taken and dried at room temperature for 3 days and then weighed. Another equal amount of lung tissue was added to 1 ml of formamide homogenate and incubated at 50°C for 24 hours to extract the dye. After centrifugation at 10,000g for 20 minutes, the supernatant containing Evans blue was collected and the absorbance was measured at a wavelength of 620 nm using a Synergy Neo2 microplate reader. The dye was quantified using a standard curve, and the pulmonary vascular permeability was finally evaluated as the lung tissue dye content / lung tissue weight (μg / g lung tissue).

[0076] Depend on Figure 5 A and D show that sepsis+ILF3 M-KO The alveolar structural damage and interstitial inflammatory infiltration in mice were significantly alleviated, and the lung injury score was significantly reduced.

[0077] Depend on Figure 5 B shows that sepsis+ILF3 M-KO The dry / wet weight ratio of lung tissue in mice was significantly lower than that in sepsis+ILF3 flox / flox group of mice.

[0078] Depend on Figure 5 C shows that knockout of ILF3 gene in macrophages significantly improved pulmonary vascular leakage caused by sepsis.

[0079] The above results show that ILF3 flox / flox 、ILF3 M-KO The alveolar structure of the lung tissue of mice was normal, no pulmonary edema occurred, and the pulmonary vascular structure was normal. flox / flox Compared with mice, the sepsis symptoms of mice with specific knockout of the ILF3 gene in macrophages were significantly alleviated.

[0080] 5. Comparison of cell apoptosis in different groups of mice

[0081] Cell apoptosis in mouse lung tissue was assessed using an in situ cell death detection kit (Roche, Basel, Switzerland) according to the manufacturer's instructions. Fluorescence images were observed using a Carl Zeiss LSM 710 laser confocal microscope, and the expression levels of Bax, Bcl-2, and cleaved caspase-3 were detected by Western blot to determine cell apoptosis. The results are shown in Table 1. Figure 6 shown.

[0082] Depend on Figure 6 As we know, ILF3 flox / flox 、ILF3M-KO There was no difference in apoptosis in the lung tissue of mice. flox / flox Compared with mice, sepsis+ILF3 M-KO Tunel staining was significantly reduced in mice.

[0083] Depend on Figure 6 B~E show that, compared with sepsis+ILF3 flox / flox Compared with the lung tissue of mice, sepsis+ILF3 M-KO The expression levels of Bax and Cleaved caspase-3 were decreased, and the expression level of Bcl-2 was increased in the lung tissues of mice.

[0084] In summary, the above results show that sepsis+ILF3 M-KO The apoptosis of mouse lung tissue was compared with that of sepsis+ILF3 flox / flox The mice showed significant improvement.

[0085] 6. Determination of inflammation levels in different groups of mice

[0086] Remove ILF3 flox / flox Group, ILF3 M-KO group, sepsis+ILF3 flox / flox group and sepsis+ILF3 M-KO The lung tissues of the mice in the two groups were then paraffin-embedded and sectioned for immunohistochemistry. At the same time, alveolar lavage fluid and plasma solutions were extracted and the levels of IL-1β, IL-6, IL-18, and TNF-α were detected using Elisa assays. Figure 7 shown.

[0087] Immunohistochemistry experiments were performed according to conventional methods. The primary antibodies used were CD68 (Abcam, ab283654), IL-6 (Abcam, ab290735), TNF-α (Abcam, ab1793), and MCP-1 (Abcam, ab315478). Biotin-conjugated secondary antibodies (ZSGB-BIO, Beijing, China) were used for incubation, and DAB solution (ZSGB-BIO) was used as the chromogen. ELISA kits (Cloud-Clone Corp.) were used to analyze the levels of IL-18, IL-1β, TNF-α, and IL-6 in plasma and bronchoalveolar lavage fluid samples, and their concentrations were calculated using a standard curve.

[0088] Depend on Figure 7 As we know, ILF3 flox / flox 、ILF3 M-KO There was no significant increase in the level of lung inflammation in mice and no statistical difference.flox / flox Compared with mice, sepsis+ILF3 M-KO The expression of CD68, IL-6, TNF-α and MCP-1 in the mice of the control group was significantly decreased.

[0089] Depend on Figure 7 From B to C, we can see that ILF3 flox / flox Group and ILF3 M-KO The inflammatory levels in the plasma and bronchoalveolar lavage fluid of the mice in the sepsis+ILF3 group were not significantly increased. flox / flox Compared with mice, the expression levels of IL-18, IL-1β, TNF-α and IL-6 in plasma and bronchoalveolar lavage fluid of mice in the sepsis+ILF3M-KO group were significantly decreased.

[0090] In summary, the above results indicate that knockout of the ILF3 gene in macrophages can significantly inhibit inflammatory cell infiltration and inflammatory response in sepsis.

[0091] Example 4

[0092] Based on the nucleotide sequence of the ILF3 gene as shown in SEQ ID NO.1, si-ILF3 as shown in SEQ ID NO.4 was designed, which can specifically interfere with the expression level of the ILF3 gene.

[0093] si-ILF3: 5'-CCUGUGUGAGAAAUCCAUUTT-3'.

[0094] Then ILF3 was isolated flox / flox ILF3 was isolated from mouse femurs and tibias and induced for 7 days in DMEM medium (Gibco, Grand Island, NY, USA) supplemented with 20 ng / ml macrophage colony-stimulating factor, 10% fetal bovine serum (FBS), and 1% penicillin / streptomycin (P / S). flox / flox Mouse bone marrow macrophages; then using Lipofectamine TM RNAiMAX kit to ILF3 flox / flox si-ILF3 was transfected into mouse bone marrow macrophages; finally, the ILF3 knockdown level was detected by Western blot and RT-PCR experiments. The results were as follows Figure 8 shown.

[0095] Depend on Figure 8 It can be seen that si-ILF3 can effectively knock down ILF3 flox / flox The expression levels of ILF3 gene and ILF3 protein in mouse bone marrow macrophages were significantly decreased, indicating that si-ILF3 can be used to prepare drugs for the treatment of sepsis.

[0096] The above results indicate that knockout of the macrophage-related gene ILF3 can significantly inhibit the development of sepsis and has a significant protective effect on the prevention and treatment of sepsis.

[0097] Finally, it should be noted that 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 above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of a reagent for detecting ILF3 gene expression levels in the preparation of a sepsis diagnostic product, characterized in that: The nucleotide sequence of the ILF3 gene is shown in SEQ NO.

1.

2. The use according to claim 1, characterized in that The reagent for detecting the expression level of human ILF3 gene includes the primer pair shown in SEQ ID NO. 2-3.

3. The use according to claim 1, characterized in that The detection sample of the reagent for detecting the expression level of human ILF3 gene is selected from cells, tissues, plasma or serum.

4. Use of the ILF3 gene as a drug target in the preparation of a drug for treating sepsis, characterized in that: The nucleotide sequence of the ILF3 gene is shown in SEQ NO.

1.

5. The use according to claim 4, characterized in that The drug for treating sepsis uses the ILF3 gene as an intervention target and can efficiently and specifically downregulate the expression level of the ILF3 gene.

6. Application of siRNA that specifically interferes with the expression level of ILF3 gene in the preparation of drugs for the treatment of sepsis.

7. The use according to claim 6, characterized in that The nucleotide sequence of the siRNA that specifically interferes with the expression level of the ILF3 gene is shown in SEQ ID NO. 4 and 5.

8. A drug for treating sepsis, comprising siRNA that specifically interferes with the expression level of the ILF3 gene.

9. The use according to claim 9, characterized in that The nucleotide sequence of the siRNA that specifically interferes with the expression level of the ILF3 gene is shown in SEQ ID NO.4.

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