Mesenchymal stem cells overexpressing il-1ra and uses thereof

Abstract

The application provides mesenchymal stem cells overexpressing IL-1Ra and application thereof, and belongs to the technical field of biological medicine. By overexpressing IL-1Ra in MSCs, the migration function of the MSCs is significantly improved, and the number of MSCs homing to the tissue damage site is significantly increased; in addition, the MSCs overexpressing IL-1Ra can also secrete IL-1Ra, so as to achieve the synergistic anti-inflammatory effect with the MSCs, thereby significantly improving the anti-inflammatory ability and tissue damage repair ability of the MSCs, and improving the treatment effect of the MSCs. The MSCs overexpressing IL-1Ra can be used for treating various local or systemic inflammatory diseases, and has good curative effect in the treatment of hemorrhagic cystitis and acute pulmonary inflammatory lesion diseases.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a mesenchymal stem cell overexpressing IL-1Ra and its applications. Background Technology

[0002] Mesenchymal stem cells (MSCs) are adult stem cells with the ability to self-renew and differentiate into various cell types, such as adipocytes, chondrocytes, osteocytes, muscle cells, and nerve cells. MSCs are mainly distributed in the interstitial tissues and organs, including bone marrow, adipose tissue, and umbilical cord. Under physiological conditions, the primary function of MSCs is to maintain the homeostasis of tissues and organs. MSCs possess paracrine functions, regulating the immune system by secreting various cytokines, promoting angiogenesis, and facilitating the regeneration and repair of cells, tissues, and organs. Furthermore, MSCs can also regulate immune cell function through direct interaction with immune cells (such as T lymphocytes, macrophages, and dendritic cells), thereby suppressing immune responses and reducing tissue and organ damage caused by excessive immune responses. Recent studies have found that exosomes derived from MSCs can promote the regeneration and repair of various tissues, including nerves and blood vessels. Given the powerful anti-inflammatory effects and biological functions of MSCs in promoting tissue repair, MSCs are widely used in clinical research or clinical trials for the treatment of inflammatory diseases such as graft-versus-host disease (GVHD), systemic lupus erythematosus, inflammatory bowel disease, pneumonia, and arthritis.

[0003] Currently, MSCs have limited ability to treat inflammatory diseases or promote tissue repair. Numerous studies have shown that the number of MSCs at the site of tissue damage is a key factor determining therapeutic efficacy, and pathological conditions and the immune environment influence MSC migration and homing functions. Therefore, finding technologies or methods to enhance MSC migration function to improve their anti-inflammatory and tissue repair capabilities is a current research hotspot.

[0004] Interleukin-1α (IL-1α) and interleukin-1β (IL-1β) are important pro-inflammatory cytokines. IL-1α and IL-1β first bind to their target cell receptor IL-1R1, and then interact with co-receptors to form a ternary complex, activating intracellular signaling pathways and inducing the expression of a series of inflammation-related molecules, leading to inflammation. Interleukin-1 receptor antagonists (IL-1Ra) can bind to IL-1R1, thereby blocking the inflammatory response induced by IL-1α and IL-1β. Anakinra is a recombinant human IL-1Ra drug that is currently used in the treatment of rheumatic diseases such as rheumatoid arthritis and some non-rheumatic inflammatory diseases.

[0005] MSCs possess characteristics such as high proliferation, ease of isolation, culture and expansion, easy transfection and expression of exogenous genes, and low immunogenicity, making them valuable for clinical applications in cell therapy and gene therapy. They are considered ideal target cells for tissue engineering, cell therapy, and gene therapy. Patent application CN116103243A (application number 202310051400.4) discloses the application of IL-1Ra overexpressing mesenchymal stem cells in the prevention and treatment of GVHD. Lentiviral viruses are prepared by packaging plasmids overexpressing IL-1Ra, which are then used to infect MSCs. After resistance selection, stable cell lines are obtained. These cell lines can be used for the prevention of aGVHD (acute graft-versus-host disease) with good preventive effects. Patent application CN117467666A (application number 202311212422.0) discloses mesenchymal stem cells overexpressing IL-1Ra and their applications. In hematopoietic stem cell transplantation, MSCs overexpressing IL-1Ra are simultaneously or sequentially infused with hematopoietic stem cells, achieving effects such as preventing GVHD, promoting hematopoietic stem cell engraftment, supporting hematopoietic stem cell proliferation, and improving the success rate of hematopoietic stem cell transplantation. In a mouse ARDS (acute respiratory distress syndrome) model, MSC exosomes overexpressing IL-1Ra showed good therapeutic effects. However, none of the aforementioned patents disclose the effect of IL-1Ra overexpression on MSC migration.

[0006] Currently, exploring a technology or method to enhance the chemotactic function of MSCs in order to improve their anti-inflammatory and tissue damage repair capabilities is of great significance to my country's medical field. Summary of the Invention

[0007] To address the shortcomings of existing technologies, this invention provides a mesenchymal stem cell overexpressing IL-1Ra and its application. By overexpressing IL-1Ra in MSCs, the migration of MSCs to sites of tissue damage can be promoted, thereby enhancing the anti-inflammatory capacity and tissue repair capacity of MSCs.

[0008] The technical solution of this invention is as follows:

[0009] A type of mesenchymal stem cell that overexpresses IL-1Ra is a mesenchymal stem cell that overexpresses IL-1Ra.

[0010] The mRNA sequence of IL-1Ra has the accession number NM_000577.5 in GenBank.

[0011] The amino acid sequence of IL-1Ra has accession numbers in GenBank as NP_776214.1, NP_776213.1, NP_000568.1, NP_776215.1, NP_001305843.1 or NP_001366289.1.

[0012] Preferably, the method for overexpressing IL-1Ra in mesenchymal stem cells is to prepare lentivirus by packaging with a plasmid containing IL-1Ra, then infect mesenchymal stem cells with the lentivirus, and screen to obtain mesenchymal stem cells overexpressing IL-1Ra.

[0013] Preferably, the mesenchymal stem cells are derived from the umbilical cord, bone marrow, fat, or dental pulp.

[0014] Preferably, the mesenchymal stem cells are derived from the umbilical cord.

[0015] The application of the mesenchymal stem cells overexpressing IL-1Ra in the preparation of drugs for treating inflammatory diseases.

[0016] Preferably, the inflammatory disease is hemorrhagic cystitis or acute pneumonia.

[0017] More preferably, the mesenchymal stem cells overexpressing IL-1Ra treat inflammatory diseases by promoting the migration of mesenchymal stem cells.

[0018] The application of mesenchymal stem cells overexpressing IL-1Ra in the preparation of tissue damage repair drugs.

[0019] The application of IL-1Ra overexpressing mesenchymal stem cells in promoting mesenchymal stem cell migration.

[0020] Preferably, the mesenchymal stem cells overexpressing IL-1Ra are used to promote the migration of mesenchymal stem cells to sites of tissue damage.

[0021] Beneficial effects:

[0022] This invention reveals that overexpressing IL-1Ra in MSCs promotes the migration of MSCs to sites of tissue damage, significantly increasing the number of MSCs at these sites. Furthermore, MSCs overexpressing IL-1Ra can secrete IL-1Ra, achieving a synergistic anti-inflammatory effect, thereby significantly enhancing the anti-inflammatory capacity and tissue repair ability of MSCs. Compared to normal MSCs, MSCs overexpressing IL-1Ra show significant therapeutic efficacy in the treatment of hemorrhagic cystitis and acute pneumonia. Attached Figure Description

[0023] Figure 1Construction and identification of MSCs overexpressing IL-1Ra. Figure A shows the recombinant expression vector overexpressing IL-1Ra; Figure B shows the expression of IL-1Ra at the mRNA level; Figure C shows the expression of IL-1Ra at the protein level; and Figure D shows the concentration of IL-1Ra secreted in the cell culture supernatant.

[0024] Figure 2 Expression of CXCR5 at the mRNA level in MSCs overexpressing IL-1Ra.

[0025] Figure 3 The migration ability of MSCs overexpressing IL-1Ra was detected using Transwell chambers. Figure A shows crystal violet-stained images of Mock-MSCs passing through the Transwell chambers; Figure B shows crystal violet-stained images of oeIL-1Ra-MSCs passing through the Transwell chambers; and Figure C shows the statistical analysis results of the number of migrating cells.

[0026] Figure 4 The therapeutic effect of MSCs overexpressing IL-1Ra on hemorrhagic cystitis. Figure A shows the hematuria status of rats in each group as detected by hematuria test strips; Figure B shows the statistical analysis results of hematuria scores; Figure C shows the gross bladder appearance of rats in each group; and Figure D shows the statistical analysis results of the ratio of bladder wet weight to body weight in rats in each group.

[0027] Figure 5 The chemotactic homing ability of MSCs overexpressing IL-1Ra to bladder tissue of rats with hemorrhagic cystitis. Figure A shows the immunohistochemical staining results of human CD105 in the bladder tissue of each group of rats; Figure B shows the statistical analysis results of the number of CD105 positive cells in the Mock-MSCs treatment group and the oeIL-1Ra-MSCs treatment group.

[0028] Figure 6 The therapeutic effect of MSCs overexpressing IL-1Ra on acute pneumonia lesions. Figure A shows the chest imaging results of mice in each group; Figure B shows the hematoxylin and eosin staining results of lung pathological sections of mice in each group. Detailed Implementation

[0029] The following description is based on specific embodiments:

[0030] Explanation of the source of experimental materials:

[0031] psPAX2: Purchased from Shanghai Jikai Gene Technology Co., Ltd.;

[0032] pMD2.G: Purchased from Shanghai Jikai Gene Technology Co., Ltd.;

[0033] Lipofectamine 3000 transfection reagent: purchased from Thermo Fisher Scientific;

[0034] opti-MEM medium: purchased from Thermo Fisher Scientific;

[0035] DMEM / F12 complete medium: 10% (10mL / 100mL) FBS, 0.272 g / L L-glutamine, 10ng / mL bFGF (fibroblast growth factor), 100U / mL penicillin, 0.1mg / mL streptomycin.

[0036] Example 1: Construction and identification of MSCs overexpressing IL-1Ra

[0037] I. Construction of MSCs overexpressing IL-1Ra (oeIL-1Ra-MSCs)

[0038] (1) Isolation, culture and amplification of MSCs

[0039] Collect umbilical cord tissue from newborns (with informed consent from the mother), add an appropriate amount of DMEM / F12 complete medium, and use sterile scissors to cut the umbilical cord tissue into small pieces as much as possible in a sterile operating table. Then, digest the tissue sequentially with 1% collagenase II solution and 0.125% trypsin solution for 30 minutes each to obtain digestion solution. Collect the digestion solution into a centrifuge tube, centrifuge at 1500 rpm for 10 minutes, discard the supernatant, and resuspend the cell pellet in an appropriate amount of DMEM / F12 complete medium to obtain a resuspended solution. Transfer the resuspended solution to a cell culture flask and incubate in a humidified, 37°C, 5% CO2 incubator. On the third day, aspirate the supernatant, remove non-adherent cells and tissue fragments, add fresh DMEM / F12 complete medium, and continue culturing. Replace the medium with fresh medium every other day thereafter.

[0040] When the cells reach 80% confluence, the supernatant is aspirated, the cells are washed once with PBS buffer, and 0.25% trypsin solution is added for digestion. After the cells become round and float, DMEM / F12 complete medium is added to terminate the reaction, yielding a digestion solution. The digestion solution is collected into a centrifuge tube, centrifuged at 1200 rpm for 5 minutes, the supernatant is discarded, and an appropriate amount of DMEM / F12 complete medium is added to the cell pellet for resuspending. The cell suspension is transferred to a cell culture flask and cultured under the incubation conditions described above. The cultured fourth-generation cells are used for subsequent experiments.

[0041] (2) Construction of oeIL-1Ra-MSCs

[0042] Using the sequence NM_000577.5 from GenBank as the target gene, the target gene was ligated into the GV367 vector (Ubi-MCS-SV40-EGFP-IRES-puromycin) to construct the recombinant expression vector oeIL-1Ra-GV367. This work was commissioned to Shanghai Jikai Gene Technology Co., Ltd. The map of the recombinant expression vector is shown below. Figure 1 As shown in Figure A.

[0043] Using 293T cells (ATCC CRL-3216) as the tool cells, oeIL-1Ra-GV367:psPAX2:pMD2.G was mixed at a mass ratio of 4:3:1. The lentivirus was packaged and transfected into 293T cells with the assistance of Lipofectamine 3000 transfection reagent. After incubation in an incubator for 48 hours, the cell supernatant was collected and centrifuged at 100,000 rpm for 2 hours. The supernatant was removed and the precipitate was resuspended in PBS buffer to obtain oeIL-1Ra lentivirus.

[0044] Viral titers were determined by infecting 293T cells with lentivirus; titers were found to be between 3 × 10⁻⁶. 8 It can be used if the concentration is above TU / mL.

[0045] The MSCs obtained in step (1) were resuspended in Opti-MEM medium, and the MSCs were then thawed at 2 × 10⁻⁶ ppm. 5 Seed cells at a density of 10 cells / well into 6-well plates, add oeIL-1Ra lentivirus at MOI=40 (multiple of infection, the ratio of viral load to cell count), incubate for 12 hours, then replace with DMEM / F12 complete medium and continue culturing; 3 days later, digest the cells according to the method described in step (1); take 1×10 5 The percentage of GFP-positive cells was measured by flow cytometry, which represents the viral infection positivity rate. Puromycin was added to the cells at a final concentration of 2 μg / mL. After 2 days, uninfected cells died and floated to the surface; the surviving cells were identified as positive cells. These positive cells were then cultured and expanded to obtain a stable MSC line overexpressing IL-1Ra: oeIL-1Ra-MSCs.

[0046] The irrelevant sequence (5'-TTCTCCGAACGTGTCACGT-3') was ligated into the GV367 vector to construct the recombinant expression vector Mock-GV367. Then, Mock lentivirus was prepared and MSCs were infected using the same method as above to obtain Mock-MSCs, which served as a blank control group for subsequent experiments.

[0047] II. Identification of oeIL-1Ra-MSCs

[0048] (1) RT-PCR identification

[0049] oeIL-1Ra-MSCs were digested with 0.25% trypsin solution, counted, and 1×10⁻⁶ samples were collected. 6 Cellular mRNA was extracted from cells and reverse transcribed into cDNA. The relative expression of IL-1Ra at the mRNA level was detected by real-time quantitative PCR (RT-PCR) using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal reference gene.

[0050] The RT-PCR primers used for detecting IL-1Ra were IL-1Ra-F and IL-1Ra-R, with primer sequences shown in SEQ ID NOs.1~2, and the product size was 167bp. The RT-PCR primers used for detecting GAPDH were GAPDH-F and GAPDH-R, with primer sequences shown in SEQ ID NOs.3~4, and the product size was 211bp.

[0051] The results are as follows Figure 1 As shown in Figure B, compared with the blank control group, the relative expression level of IL-1Ra at the mRNA level in the stable cell line oeIL-1Ra-MSCs was significantly increased.

[0052] (2) Western blot identification

[0053] Take 1×10 6 One oeIL-1Ra-MSC was lysed with RIPA lysis buffer to prepare protein samples. GAPDH was used as an internal control protein, and the expression level of IL-1Ra was detected by Western blot. Protein concentration was determined using the BCA method. Equal amounts of protein samples were separated by SDS-PAGE gel electrophoresis and transferred to a PVDF membrane. The PVDF membrane was blocked with 5% BSA at room temperature for 1 hour, and anti-human IL-1Ra antibody was added and incubated overnight at 4°C. The membrane was washed with TBST, and horseradish peroxidase-labeled secondary antibody was added and incubated at room temperature for 1 hour. After washing with TBST, the membrane was developed using a chemiluminescent substrate.

[0054] The results are as follows Figure 1 As shown in Figure C, compared with the blank control group, the relative expression level of IL-1Ra in the stable cell line oeIL-1Ra-MSCs was significantly increased.

[0055] (3) ELISA identification

[0056] oeIL-1Ra-MSCs were cultured in DMEM / F12 complete medium at 37°C and 5% CO2 for 72 hours. The cell culture supernatant was collected. The OD of the supernatant was measured using a microplate reader according to the ELISA kit instructions. 450 OD 630 Plot a standard curve, and then calculate the concentration of IL-1Ra in the supernatant based on the standard curve.

[0057] The results are as follows Figure 1 As shown in Figure D, compared with the blank control group, the concentration of IL-1Ra in the cell culture supernatant of the stable cell line oeIL-1Ra-MSCs was significantly increased, which indicates that MSCs overexpressing IL-1Ra can secrete IL-1Ra into the extracellular space.

[0058] (4) Expression of chemokine CXCR5

[0059] The relative expression of chemokine CXCR5 in oeIL-1Ra-MSCs was detected by RT-PCR. The RT-PCR primers used were CXCR5-F and CXCR5-R, and the primer sequences are shown in SEQ ID NOs.5~6, respectively. The product size was 233bp.

[0060] The results are as follows Figure 2 As shown. By Figure 2 The relative expression level of CXCR5 at the mRNA level in oeIL-1Ra-MSCs was significantly higher than that in the blank control group, suggesting that overexpression of IL-1Ra may enhance the chemotactic ability of MSCs by upregulating the expression of CXCR5.

[0061] In summary, compared with the blank control group, the expression levels of IL-1Ra in the stable cell line oeIL-1Ra-MSCs were significantly increased at both the mRNA and protein levels, and the concentration of IL-1Ra in the cell culture supernatant was significantly increased, indicating that the MSCs overexpressing IL-1Ra were successfully constructed.

[0062] Example 2: Migration ability test of MSCs overexpressing IL-1Ra

[0063] oeIL-1Ra-MSCs were resuspended in FBS-free DMEM / F12 complete medium to achieve a cell concentration of 1×10⁻⁶. 5 Cells / mL; meanwhile, Mock-MSCs were used as a blank control group.

[0064] Chemotaxis assays were performed using 24-well plate chemotaxis chambers with 8 μm pore size. 100 μL of the cell resuspension solution was added to the upper chamber, and 600 μL of DMEM / F12 complete culture medium was added to the lower chamber. After 24 hours, excess liquid in the upper chamber was removed, and unmigrated cells were wiped off the inner side of the upper chamber membrane with a cotton swab. The upper chamber was fixed with 4% paraformaldehyde solution for 20 minutes, and then the outer side of the upper chamber membrane was stained with 0.5% crystal violet for 10 minutes. Cell migration was observed in five random fields of view under a microscope, and the number of migrating cells was counted.

[0065] The results are as follows Figure 3 As shown, figures A and B depict cell migration on the outer side of the superior chamber membrane, while figure C presents the statistical results of the number of migrating cells in each group. Figure 3 It was found that, compared with the blank control group, the number of migrating oeIL-1Ra-MSCs was significantly increased (****) p <0.0001), which indicates that MSCs overexpressing IL-1Ra have significantly enhanced chemotactic ability.

[0066] Application Example 1: Treatment of hemorrhagic cystitis with MSCs overexpressing IL-1Ra

[0067] I. Construction of a rat model of hemorrhagic cystitis

[0068] Twenty-four male Wistar rats (250-280g, 6 weeks old) were selected, of which 6 rats served as the normal control group (control, con) without any treatment; the remaining 18 rats were injected intraperitoneally with 40mg / mL cyclophosphamide solution on day 0, with an injection dose of 150mg / kg, to establish a rat model of hemorrhagic cystitis.

[0069] Eighteen rats with hemorrhagic cystitis were randomly divided into three groups of six each: a disease model group (hemorrhagic cystitis), a Mock-MSCs treatment group, and an oeIL-1Ra-MSCs treatment group. The disease model group received no intervention. The Mock-MSCs treatment group received tail vein infusion of Mock-MSCs on days 1 and 3, respectively, while the oeIL-1Ra-MSCs treatment group received tail vein infusion of oeIL-1Ra-MSCs on days 1 and 3, respectively. The infusion dose for both groups was 1×10⁻⁶. 6 Each / each

[0070] II. Observation of Treatment Effect

[0071] (1) Evaluation of blood and urine test strips

[0072] On day 7, urine samples were collected from each rat using blood and urine test strips. The results were compared and scored according to the standard colorimetric spectrum of the blood and urine test strips, and statistical analysis was performed.

[0073] The blood urine test strips showed the following in rats: Figure 4 As shown in Figure A, the hematuria score results are as follows: Figure 4 As shown in Figure B (** p <0.01, *** p <0.001, **** p <0.0001). As shown in Figures A and B, compared with the Mock-MSCs treatment group, the hematuria in the rats in the oeIL-1Ra-MSCs treatment group was significantly alleviated.

[0074] (2) Observation and weighing of rat bladder tissue

[0075] Rats were euthanized, and bladder tissue was collected for photographing and weighing (wet weight) to observe changes in the bladder tissue.

[0076] Photographs of bladder tissue, such as Figure 4 As shown in Figure C, the statistical analysis results of the ratio of bladder tissue wet weight to body weight are as follows: Figure 4 As shown in Figure D (**) p <0.01, **** p <0.0001). As shown in Figures C to D, compared with the Mock-MSCs treatment group, the rats in the oeIL-1Ra-MSCs treatment group had less bladder bleeding, smaller bladder volume, and significantly lower wet weight.

[0077] (3) Chemotaxis and homing ability of oeIL-1Ra-MSCs to rat bladder tissue

[0078] Bladder tissue was fixed with 4% paraformaldehyde solution, embedded in paraffin, and 8 mm bladder tissue sections were prepared. The bladder tissue sections were stained with hematoxylin and eosin according to the standard method, and human CD105 cells (a marker molecule of mesenchymal stem cells) were labeled by immunohistochemistry to observe the chemotactic homing ability of oeIL-1Ra-MSCs to rat bladder tissue.

[0079] The results are as follows Figure 5 As shown in the figure, Figure A shows the immunohistochemical staining results of human CD105 in rat bladder tissue of each group, and Figure B shows the statistical analysis results of the number of CD105 positive cells (light brown, larger cells) in each group (*). p <0.05). By Figure 5 It was found that oeIL-1Ra-MSCs were significantly more localized in the bladder tissue of a rat model of hemorrhagic cystitis.

[0080] In summary, MSCs overexpressing IL-1Ra were more effective than the control group (Mock-MSCs treatment group) in treating hemorrhagic cystitis, and MSCs overexpressing IL-1Ra showed a significantly increased chemotactic ability toward the site of inflammatory cystitis.

[0081] Application Example 2: Treatment of acute pneumonia lesions with MSCs overexpressing IL-1Ra

[0082] I. Construction of a mouse model of acute pneumonia

[0083] Lipopolysaccharide (LPS) was dissolved in PBS buffer to prepare an LPS solution with a concentration of 2 mg / mL.

[0084] Twenty-four male Balb / c mice (20-22g, 6-8 weeks old) were selected and acclimatized for one week. Six mice served as the normal control group, while the remaining 18 mice were intratracheally instilled with LPS solution at a dose of 5 mg / kg to establish an acute pneumonia mouse model. The normal control group was instilled with an equal volume of PBS buffer.

[0085] Eighteen mice with ALI were randomly divided into three groups of six each: an LPS model group, a Mock-MSCs treatment group, and an oeIL-1Ra-MSCs treatment group. In the Mock-MSCs treatment group, Mock-MSCs cells were instilled into the trachea 4 hours after LPS infusion, while in the oeIL-1Ra-MSCs treatment group, oeIL-1Ra-MSCs cells were instilled into the trachea 4 hours after LPS infusion. The infusion dose for both groups was 5 × 10⁻⁶ cells. 5 Each individual; the LPS model group was instilled with an equal volume of PBS buffer.

[0086] II. Observation of Treatment Effect

[0087] (1) Chest imaging examination

[0088] Small animal micro-CT technology was used to perform chest imaging examinations on mice in each group to assess the degree of lung lesions. The results are as follows: Figure 6 As shown in Figure A.

[0089] Depend on Figure 6 As shown in Figure A, the lungs of mice in the normal control group had normal brightness and no obvious abnormalities; patchy exudative shadows were visible in the lungs of mice in the LPS model group, and some showed actual lesions; the inflammatory lesions in the lungs of mice in the Mock-MSCs treatment group were reduced compared with those in the LPS model group; compared with the Mock-MSCs treatment group, the inflammatory lesions in the lungs of mice in the oeIL-1Ra-MSCs treatment group were reduced more significantly, and there were no obvious exudative shadows.

[0090] (2) Observation of pathological sections

[0091] Pathological sections of the lungs of mice in each group were prepared and stained with hematoxylin and eosin. The degree of lung lesions was then observed under a microscope. The results are as follows: Figure 6 As shown in Figure B.

[0092] Depend on Figure 6 As shown in Figure B, the lungs of mice in the normal control group showed a small number of inflammatory cells, full alveoli, thin and intact alveolar walls, and no exudation in the alveolar cavities. The lungs of mice in the LPS model group showed full-field inflammatory cells, widened pulmonary interstitium, alveolar collapse, exudation in the alveolar cavities, and pulmonary capillary congestion and hemorrhage. The lung edema, hemorrhage, and congestion of mice in the Mock-MSCs treatment group were reduced compared to the LPS model group. Compared to the Mock-MSCs treatment group, the reduction in lung edema, hemorrhage, and congestion was more significant in the oeIL-1Ra-MSCs treatment group.

[0093] In summary, MSCs overexpressing IL-1Ra can effectively treat acute pneumonia.

Claims

1. Use of mesenchymal stem cells overexpressing IL-1Ra, characterized in that, The application is applied to the preparation of a medicine for treating hemorrhagic cystitis, which promotes the migration of mesenchymal stem cells overexpressing IL-1Ra to the site of tissue injury; wherein the mesenchymal stem cells overexpressing IL-1Ra are mesenchymal stem cells overexpressing IL-1Ra.

2. Use according to claim 1, wherein The method for overexpressing IL-1Ra in mesenchymal stem cells is to prepare a lentivirus by using a plasmid containing IL-1Ra, then infect the mesenchymal stem cells with the lentivirus, and obtain the mesenchymal stem cells overexpressing IL-1Ra after screening.

3. The use according to claim 1, wherein The source of the mesenchymal stem cells is umbilical cord, bone marrow, fat or dental pulp.

4. The use according to claim 1, wherein The source of the mesenchymal stem cells is umbilical cord.

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