Use of rage receptor inhibitor in preparation of drug for inhibiting lymphocyte reduction or death

Abstract

Disclosed is use of a Rage receptor inhibitor in the preparation of a drug for inhibiting lymphocyte reduction or death. Using Rage as a target for in vitro screening of the drug for inhibiting lymphocyte reduction or death provides a new research direction for preparing more efficient tumor drugs.

Description

Application of Rage receptor inhibitors in the preparation of drugs that inhibit lymphopenia or lymphocyte death Technical Field

[0001] This application belongs to the field of biotechnology, specifically relating to the application of Rage receptor inhibitors in the preparation of drugs that inhibit lymphocyte reduction or death. Background Technology

[0002] The incidence and mortality rates of malignant tumors and severe infections are on the rise, becoming the second and third leading causes of death globally, respectively. One common characteristic of malignant tumors and severe infections is lymphocyte depletion, leading to persistent immunosuppression or tumor immune escape. Reducing lymphocyte depletion, enhancing lymphocyte activity, and promoting the body's immune response can help improve overall survival. Because immunotherapy can enhance the body's immune capacity and has the potential for high specificity and low side effects, it shows great promise for future development.

[0003] Immune checkpoint inhibitors (ICIs) work by blocking the immunosuppressive ligand-receptor interactions involving CTLA-4 and PD-1. Immune checkpoint blockade (ICB), a representative of tumor immunotherapy, has revolutionized cancer treatment. While ICB treatment reduces major suppressor signals in T lymphocytes and enhances their potential T-cell-mediated anti-tumor capabilities, it is also accompanied by progressive exhaustion and death of tumor-reactive T cells, leading to widespread drug resistance and an inability to exert a sustained effect, thus impacting treatment efficacy.

[0004] For overall clinical treatment efficacy, it is necessary to improve the response rate and duration of the body's immune function to achieve better results. Therefore, specific small molecule inhibitors may not only inhibit key oncogenic signaling pathways but also maintain cell viability while enhancing lymphocyte response, thus playing a more durable role and serving as adjunctive therapy to existing immune checkpoint inhibitors. Current ICB therapy is only effective for a small number of cancer patients, and many patients cannot benefit from treatment long-term. Therefore, the scientific community urgently needs to find mechanisms to enhance immune cell viability and avoid immune cell depletion, relieve immunosuppression, and thus research new target immune checkpoint inhibitors, combining immune checkpoint inhibitors with different targets, or combining immune checkpoint inhibitors with other treatment methods to enhance immune cell viability and reduce immune cell depletion.

[0005] Summary of the Invention

[0006] This application aims to at least address one of the technical problems existing in the prior art described above. To this end, this application proposes the use of Rage as a target in the in vitro screening of products that inhibit lymphopenia or lymphocyte death.

[0007] This application also proposes the use of Rage receptor inhibitors in the preparation of products that inhibit lymphopenia or lymphocyte death.

[0008] This application also proposes a method for screening drugs that inhibit lymphopenia or lymphocyte death.

[0009] According to the first aspect of this application, the use of Rage as a target in the in vitro screening of products that inhibit lymphopenia or lymphocyte death is proposed.

[0010] In some embodiments of this application, the lymphocytes include T lymphocytes and B lymphocytes.

[0011] According to a second aspect of this application, the use of Rage receptor inhibitors in the preparation of products that inhibit lymphopenia or lymphocyte death is proposed.

[0012] In some embodiments of this application, the Rage receptor inhibitor refers to a molecule that can inhibit the transcription or translation of the Rage gene, or a molecule that can specifically inhibit the expression or activation of the Rage protein.

[0013] In some embodiments of this application, the Rage receptor inhibitor is a nucleic acid molecule, an antibody, or a small molecule compound.

[0014] In some embodiments of this application, the nucleic acid molecule is selected from siRNA, shRNA, or sgRNA.

[0015] In some embodiments of this application, the small molecule compound is selected from one or both of FPS-ZM1 and Azirige.

[0016] In some embodiments of this application, the product that inhibits lymphocyte reduction or death has any one of the following functions (1) to (3):

[0017] (1) Increase CD3 + T cell count;

[0018] (2) Increase CD19 + B cell count;

[0019] (3) Inhibit programmed cell death.

[0020] In some embodiments of this application, the programmed cell death is pyroptosis, apoptosis, or necroptosis.

[0021] In some embodiments of this application, the Rage receptor inhibitor may be used alone or in combination with other drugs.

[0022] In some embodiments of this application, the product that inhibits lymphocyte reduction or death includes a single Rage receptor inhibitor or a combination of the Rage receptor inhibitor with other drugs.

[0023] In some embodiments of this application, the Rage receptor inhibitor is combined with pharmaceutically acceptable excipients to form a pharmaceutical composition.

[0024] In some embodiments of this application, the pharmaceutically acceptable excipient is selected from one or more of carriers, diluents, binders, lubricants, and wetting agents.

[0025] In some embodiments of this application, the pharmaceutical composition is selected from one or more of the following forms: solution, injection, spray, nasal drops, aerosol, powder, tablet, capsule, and granule.

[0026] In some embodiments of this application, the pharmaceutical composition may be introduced into the body, such as into muscles, intradermal, subcutaneous, venous, or mucosal tissues, by means of injection, spray, nasal drops, eye drops, penetration, absorption, or physical or chemical mediation; or it may be introduced into the body after being mixed or encapsulated with other substances.

[0027] According to a third aspect of this application, a method for screening drugs that inhibit lymphopenia or lymphocyte death is proposed, the method comprising: using Rage as a drug target, searching for substances that can inhibit or block the expression and / or function of Rage as candidate drugs.

[0028] In some embodiments of this application, the method includes: applying a candidate drug to a cell or animal model in vitro, and detecting the reduction or death of lymphocytes in the cell or animal model after co-culturing.

[0029] In some embodiments of this application, the cells may be derived from mammals.

[0030] Researchers can determine whether a drug has therapeutic significance by detecting the reduction or death of lymphocytes after co-culture. Generally speaking, drugs that reduce the reduction or death of lymphocytes by 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% respectively compared to the control group can be considered therapeutically significant.

[0031] In this application, treatment refers to slowing down, interrupting, preventing, controlling, stopping, alleviating, reducing, or reversing the progression or severity of a sign, symptom, disorder, condition, or disease after the disease has begun to develop, but does not necessarily involve the complete elimination of all disease-related signs, symptoms, conditions, or disorders.

[0032] In this application, prevention refers to all actions that suppress or delay symptoms by applying the product described in this application.

[0033] In some embodiments of this application, the drug is deemed to be therapeutically significant if it can reduce lymphocyte reduction or death by at least 50%.

[0034] According to some embodiments of this application, at least the following beneficial effects are achieved: This application creatively uses Rage as a target to screen drugs that inhibit lymphocyte reduction or death in vitro, providing a new research direction for the preparation of more efficient tumor drugs. Attached Figure Description

[0035] The present application will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0036] Figure 1 is a schematic diagram of the experimental operation of Embodiment 1 of this application;

[0037] Figure 2 shows the upper chamber Rage of Embodiment 1 of this application. + / + or Rage - / - Flow cytometry results of T and B lymphocyte survival rate and number in spleen cells, where CD3-FITC, CD4-AF700, and CD8-PE represent T lymphocytes, and CD19-APC / Cy7 represent B lymphocytes; Rage + / + S and Rage - / - S Indicates the lower room Rage + / + PM Different genotypes of upper ventricular spleen cells were co-cultured for 72 h; Vehicle represents an equal volume control group, and IFNγ (50 ng / mL) is the experimental group; the numbers in red font represent the proportion of lymphocytes in the cell count.

[0038] Figure 3 is a statistical graph of T and B lymphocyte survival in the spleen cells of the upper chamber corresponding to the flow cytometry detection results of Example 1 of this application and Figure 2; where %Vehicle represents Rage + / + S and Rage - / - S The ratio of lymphocyte count in the experimental group to the lymphocyte count in their respective control groups; A represents Rage. + / + Sand Rage - / - S CD3 in the experimental group + The graph shows the ratio of T lymphocyte count to the lymphocyte count in their respective control groups. B represents Rage. + / + S and Rage - / - S CD4 in the experimental group + The graph shows the ratio of T lymphocyte count to the lymphocyte count in their respective control groups. C represents Rage. + / + S and Rage - / - S CD8 in the experimental group + The graph shows the ratio of T lymphocyte count to the lymphocyte count in their respective control groups, where D represents Rage. + / + S and Rage - / - S CD19 in the experimental group + The graph shows the ratio of T lymphocyte count to the lymphocyte count in their respective control groups. * indicates p < 0.05, and ** indicates p < 0.01.

[0039] Figure 4 shows the CD3+ levels detected by flow cytometry in the endotoxemia model of Example 2 of this application. + T lymphocyte count statistics, where * indicates p < 0.05, ** indicates p < 0.01, and **** indicates p < 0.0001;

[0040] Figure 5 shows the CD4+ levels detected by flow cytometry in the endotoxemia model of Example 2 of this application. + T lymphocyte count statistics, where ** indicates p < 0.01, and **** indicates p < 0.0001;

[0041] Figure 6 shows the CD8+ levels detected by flow cytometry in the endotoxemia model of Example 2 of this application. + T lymphocyte count statistics, where * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001;

[0042] Figure 7 shows the CD3 content of spleen tissue in the endotoxemia model of Example 2 of this application. + Immunohistochemical image of T lymphocytes;

[0043] Figure 8 shows the CD3+ values ​​detected by flow cytometry in the cecal ligation and perforation model of Example 2 of this application. + T lymphocyte percentage chart, where ** indicates p < 0.01, *** indicates p < 0.001, and **** indicates p < 0.0001;

[0044] Figure 9 shows the CD4 count obtained by flow cytometry in the cecal ligation and perforation model of Example 2 of this application.+ T lymphocyte percentage chart, where ** indicates p < 0.01, *** indicates p < 0.001, and **** indicates p < 0.0001;

[0045] Figure 10 shows the CD8+ values ​​detected by flow cytometry in the cecal ligation and perforation model of Embodiment 2 of this application. + T lymphocyte percentage chart, where * indicates p < 0.05 and ** indicates p < 0.01;

[0046] Figure 11 shows the CD19 levels detected by flow cytometry in the cecal ligation and perforation model of Embodiment 2 of this application. + A statistical chart of the proportion of B lymphocytes, where *** indicates p < 0.001 and **** indicates p < 0.0001;

[0047] Figure 12 shows the results of Western blotting detection of programmed cell death proteins in Example 2. Detailed Implementation

[0048] The following will clearly and completely describe the concept and technical effects of this application in conjunction with embodiments, so as to fully understand the purpose, features and effects of this application. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are all within the scope of protection of this application.

[0049] Experimental materials: Rage - / - Rage gene knockout mice with C57 background, Rage + / + and Rage + / - Wild-type littermate control mice for this gene knockout mouse (given to Southern Medical University; specific construction and identification methods are referenced in "RAGE Control of Diabetic Nephropathy in a Mouse Model: Effects of RAGE Gene Disruption and Administration of Low-Molecular Weight Heparin") were used, and the experimental mice were 8-12 weeks old.

[0050] Example 1: Application of Rage receptor inhibitors in the preparation of products that inhibit lymphopenia or lymphocyte death

[0051] This embodiment provides the application of Rage receptor inhibitors in the preparation of products that inhibit lymphocyte reduction or death, specifically using Rage knockout mice for subsequent experimental verification.

[0052] Experimental Method: The protocol is shown in Figure 1, and can be summarized as follows: Rage in the lower chamber of the Transwell was pre-stimulated with IFNγ at a final concentration of 50 ng / mL or an equal volume of opti-MEM (Vehicle). + / + peritoneal macrophages (Rage) + / + PM After 12 hours, the lower chamber culture medium was discarded and replaced with an equal volume of 1640 complete culture medium. After another 12 hours of culturing, spleen cells (Rage cells) were added to the upper chamber of the Transwell. + / + or Rage - / - spleen cells (Rage) + / + S or Rage - / - S After co-culturing cells in the upper and lower chambers of Transwell for 72 hours, the upper chamber Rage was detected by flow cytometry. + / + or Rage - / - The survival rate and number of various lymphocytes in spleen cells are determined through the following steps:

[0053] (1) Extraction of peritoneal macrophages from wild-type mice:

[0054] 1) Preparation:

[0055] For Rage infants aged 8-12 weeks + / + Mice (10-week-old mice in this example) were intraperitoneally injected with 3% mercaptoacetate: The injection site and surrounding area were disinfected (the injection site is ideally chosen at the midpoint of the line connecting the midline of the abdomen and the groin, where there are fewer abdominal organs and the risk of puncture is lower). Holding the syringe in the right hand (drawing 2.5 mL of 3% mercaptoacetate), with the bevel facing upwards, the syringe was slowly inserted 2 mm into the skin at a slight angle. The needle was then inserted labyrinthically at a 45° angle to the abdomen into the abdominal cavity. A distinct feeling of loss of resistance was felt, and no blood return was observed upon aspiration. Broth was slowly injected into the abdominal cavity, and the needle was then withdrawn. The mice were returned to their cages, and their condition was observed. If the injection had been administered into a blood vessel, the mouse might have died immediately.

[0056] 2) Cell Extraction: Three days later, mice were euthanized by cervical dislocation and completely immersed in alcohol for 5 minutes for disinfection. 15 mL of 1640 culture medium was drawn up using a 20 mL syringe. The laminar flow hood was opened, and the mouse was removed and placed on sterilized kitchen paper, head to the left, in a supine position. The skin of the lower abdomen was gently lifted with curved forceps, and an incision of approximately 2-3 mm was made along the forceps with ophthalmic scissors. The skin was gently torn open to fully expose the peritoneum. The peritoneum was lifted with curved forceps in the left hand, and the 20 mL syringe containing the 1640 culture medium was immediately injected into the peritoneum to flush out the greater omentum and intestines adjacent to the abdominal wall. Holding the syringe with the bevel facing the user, the peritoneum was repeatedly flushed back and forth 5-8 times. The 1640 culture medium was observed to become cloudy. The peritoneal lavage fluid was collected in a 50 mL sterile centrifuge tube.

[0057] 3) Cell Counting and Plating: Transfer all cells to a clean bench and filter using a cell strainer to remove fatty tissue and other impurities from the washing fluid. Centrifuge at 129×g for 5 min at 4℃. Discard the supernatant and add 2 mL of preheated 1640 complete culture medium. Pipette the cells approximately 50 times. Transfer 10 μL of the cell suspension to 390 μL of 1640 complete culture medium and repeat the pipetting. Transfer 10 μL of the suspension to a counting chamber using a micropipette and count the cells under a microscope.

[0058] Count the total number of cells in the two diagonally opposite large squares on the counting plate, and then calculate the mean. The resulting cell count is the mean number of cells in the two large squares per milliliter of cell suspension × 10. 4 .

[0059] Cell seeding: After cell counting, transfer the cells to the appropriate culture plates according to the seeding status. The experimental plates are 24-well plates, with 500 μL of cell suspension per well, and a cell density of 5 × 10⁶ cells per 500 μL of 1640 complete culture medium. 5 1640 cells (complete culture medium: fetal bovine serum containing 10% inactivated complement + 1% penicillin / streptomycin + 10 μmol / L β-mercaptoethanol).

[0060] 4) Rage was prestimulated with mouse recombinant protein IFNγ at a final concentration of 50 ng / mL. + / + Peritoneal macrophages were cultured for 12 hours, after which the cell culture supernatant was discarded and replaced with fresh 1640 complete culture medium. The cells were then returned to the cell culture incubator for another 12 hours. Rage cells treated as described above... + / + peritoneal macrophages (Rage + / + PM This serves as the lower chamber of the Transwell. The control vehicle had its recombinant protein IFNγ replaced with an equal volume of opti-MEM prestimulated Rage. + / + Peritoneal macrophages.

[0061] (2) Extracting Rage - / - Mice and Rage + / + Spleen cells of mice:

[0062] 1) Take Rage - / - Mice and age- and sex-matched Rage + / + Mouse spleen, single spleen cell suspension prepared using a 40μm filter;

[0063] 2) Centrifuge the single spleen cell suspension to settle the cells. The centrifugation conditions are 4℃, 500×g, and 5 minutes.

[0064] 3) Discard the supernatant after centrifugation, add 1× red blood cell lysis buffer (5 mL / spleen), and lyse the red blood cells at room temperature for 5 to 8 minutes;

[0065] 4) Wash the cells with 5 mL of 1×PBS on ice, then centrifuge again at 4°C, 500×g for 5 minutes, repeating 3 times.

[0066] 5) Discard the washed PBS and resuspend the washed cells in 2 mL of 1640 complete medium (1640 complete medium: fetal bovine serum containing 10% inactivated complement + 1% penicillin / streptomycin + 10 μmol / L β-mercaptoethanol);

[0067] 6) Filter the resuspended cells through a 40μm filter;

[0068] 7) Cell counting and assessment of cell viability using 0.4% trypan blue; cell viability must be ≥85% before proceeding to the next step.

[0069] 8) Use the Rage extracted and processed in (1) + / + Mouse peritoneal macrophages were used as the lower chamber of the Transwell, and Rage cells were added to the upper chamber of the Transwell. - / - mice or Rage + / + Mouse spleen cells (5 × 10⁶ cells per well) 5 Continue culturing for 72 hours (50 μL / piece);

[0070] 9) Collect upper chamber spleen cells from Transwell and stain them with CD3-FITC, CD4-AF700, CD8-PE, and CD19-APC / Cy7. Analyze the upper chamber Rage data by flow cytometry. - / - mice or Rage + / + Survival status of T and B lymphocytes in mouse spleen cells.

[0071] The results are shown in Figure 2-3. It can be seen from the figure that under the same processing conditions, Rage... - / - SThe survival rate and number of T and B lymphocytes in group A were significantly higher than those in group B. + / + S The study demonstrates that Rage gene knockout can inhibit the depletion of T and B lymphocytes. The Rage target identified in this application can be used to maintain T and B lymphocyte viability and inhibit their depletion. The screening method described in this application can be effectively used to screen drugs that inhibit T and B lymphocyte depletion.

[0072] Example 2: Application of Rage receptor inhibitors in the preparation of reagents for inhibiting programmed cell death

[0073] This embodiment demonstrates through in vivo animal experiments that blocking Rage can inhibit programmed cell death (pyroptosis, apoptosis, and necroptosis). The specific steps are as follows: 1. Verification experiment on inhibiting lymphopenia or cell death.

[0074] (1) Animal in vivo experiments: Endotoxemia mouse model.

[0075] 1) Experimental Groups: Rage + / + -NS group, Rage + / + -LPS group, Rage + / - -LPS group and Rage - / - -LPS group, where LPS represents intraperitoneal injection of 8 mg / kg lipopolysaccharides (LPS), and NS (Normal Saline) represents intraperitoneal injection of an equal volume of physiological saline; Rage + / + This means not knocking out the Rage gene, Rage + / - Represents a single knockout of the Rage gene, Rage - / - This represents a double knockout of the Rage gene. Each group should contain at least 3 mice (8-12 weeks old; 10-week-old mice were used in this example).

[0076] 2) Mice were weighed, and control mice were intraperitoneally injected with 100 μL of physiological saline. + / + -LPS, Rage + / - -LPS and Rage - / - - The LPS group received intraperitoneal injections of LPS at a final concentration of 8 mg / kg, which was equal in volume to that of normal saline.

[0077] 3) 72 hours after modeling, the spleen of the mouse was taken and ground to prepare a single spleen cell suspension (spleen taking method is the same as in Example 1) to analyze the survival of lymphocytes.

[0078] The results of the endotoxemia mouse model are shown in Figures 4-7. As can be seen from the figures, the Rage [of mice] after intraperitoneal injection of LPS [experienced significant changes]. - / - and Rage + / - The number of surviving T and B lymphocytes in group A was significantly higher than that in group Rage. + / + This indicates that knockout or knockdown of the Rage gene can inhibit the depletion of T and B lymphocytes.

[0079] (2) Animal in vivo experiments: Cecal ligation and perforation (CLP) model.

[0080] 1) Experimental Groups: Rage + / + -Sham Group, Rage - / - -Sham Group, Rage + / + -CLP Group and Rage - / - -CLP group, where Sham represents the sham surgery group; CLP represents the mouse cecal ligation and perforation surgery group.

[0081] 2) CLP modeling steps:

[0082] ①Based on the mouse's weight, the mice were anesthetized with 1% sodium pentobarbital, and the skin of each group of mice was prepared with a razor.

[0083] ② Fix the mouse's limbs, place it on its abdomen, and disinfect it with alcohol and iodine.

[0084] ③ Make a cut in the skin slightly below the midline of the abdomen, and make a longitudinal incision of less than 1 cm. Use scissors to bluntly separate the skin and rectus abdominis muscle.

[0085] ④ Cut open the rectus abdominis muscle to expose the abdominal contents;

[0086] ⑤ Use curved forceps to lift the skin and rectus abdominis muscle of the left abdomen, use straight forceps to gently explore the position of the cecum, gently remove it and place it on gauze in the correct direction;

[0087] ⑥ Ligation of the cecum: Ligate the distal end of the cecum (the ligation length is 35% of the total length of the cecum);

[0088] ⑦ Cecal perforation: Determine whether there is feces at the distal end of the cecum. If there is no feces, gently push the contents of the cecum from the proximal end to the end of the cecum. Use a 10ml syringe needle to make a single hole at the end of the cecum, avoiding blood vessels. Squeeze out some feces from the hole, wipe it away with a cotton swab, and then squeeze out a sesame seed-sized amount of feces.

[0089] ⑧ Close the abdominal cavity: Lift the right abdominal muscles and the right skin, and use straight forceps to push the intestines back in. Be careful to open and expose the right side as much as possible, and do not let feces get on the incision.

[0090] ⑨ Postoperative fluid resuscitation and recovery: Subcutaneous injection of 37°C preheated saline (5ml / 100g, the amount used in this example is about 1ml / mouse) was injected into both sides of the back. The mice were then placed on a heating pad to keep them warm. The mice's awakening indicated that the model was successfully established.

[0091] ⑩ Seven days after modeling, the spleen of the mouse was taken and ground to prepare a single spleen cell suspension (spleen taking method is the same as in Example 1) to analyze lymphocyte death.

[0092] Note: The procedure was the same for the Sham sham surgery group, but the cecum was not ligated or perforated.

[0093] The results of the CLP model are shown in Figures 8 to 11. It can be seen from the figures that Rage... - / - The survival rates of T and B lymphocytes in the CLP group were significantly higher than those in the Rage group. + / + Group. This indicates that blocking Rage significantly reduces T and B lymphocyte death.

[0094] 2. Blocking Rage can be used in experiments to inhibit programmed cell death.

[0095] Western blotting was used to detect the effects of blocking Rage in an animal model of endotoxemia (i.e., detecting Rage). + / + and Rage - / - The pyroptosis, apoptosis, and necroptosis of spleen cells in an animal model of endotoxemia.

[0096] The Western Blot results are shown in Figure 12. As can be seen from the figure, blocking Rage can significantly reduce programmed cell death, such as pyroptosis, apoptosis, and necroptosis.

[0097] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the protection of this application. Furthermore, unless otherwise specified, the embodiments and features described in the embodiments of this application can be combined with each other.

Claims

1. Use of Rage as a target in screening a product for inhibiting lymphocyte reduction or death in vitro.

2. Use according to claim 1, characterized in that, The lymphocytes include T lymphocytes and B lymphocytes.

3. Use of a Rage receptor inhibitor in the preparation of a product for inhibiting lymphocyte reduction or death.

4. Use according to claim 3, characterized in that, The Rage receptor inhibitor is a molecule capable of inhibiting the transcription or translation of the Rage gene, or a molecule capable of specifically inhibiting the expression or activation of the Rage protein; Preferably, the Rage receptor inhibitor is a nucleic acid molecule, an antibody, or a small molecule compound; More preferably, the nucleic acid molecule is selected from siRNA, shRNA, or sgRNA; More preferably, the small molecule compound is selected from one or both of FPS-ZM1 and azirugin.

5. Use according to claim 3, characterized in that, The product for inhibiting lymphocyte reduction or death has any one of the following (1) to (3) functions: (1) increasing the content of CD3 + T cells; (2) increase CD19 + B cell content; (3) inhibiting programmed cell death; Preferably, the programmed cell death is pyroptosis, apoptosis, or necroptosis.

6. Use according to claim 3, characterized in that, The product for inhibiting lymphocyte reduction or death includes a single Rage receptor inhibitor or a combination of the Rage receptor inhibitor and other drugs.

7. Use according to claim 3, characterized in that, The Rage receptor inhibitor is combined with a pharmaceutically acceptable excipient to form a pharmaceutical composition; preferably, the pharmaceutically acceptable excipient is selected from one or more of a carrier, a diluent, a binder, a lubricant, and a wetting agent.

8. Use according to claim 7, characterized in that, The form of the pharmaceutical composition is selected from one or more of a solution, an injection, a spray, a nose drop, an aerosol, a powder spray, a tablet, a capsule, and a granule.

9. A method of screening for a drug that inhibits lymphocyte depletion or death, characterized by, The method includes using Rage as a drug target to find a substance capable of inhibiting or blocking the expression and / or function of Rage as a candidate drug.

10. The method of claim 9, wherein, The method includes applying the candidate drug to cells or animal models in vitro, and detecting the reduction or death of lymphocytes in the cells or animal models after co-culture.

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