CN117357632A - Application of IL-33 recombinant protein - Google Patents
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- CN117357632A CN117357632A CN202311436301.4A CN202311436301A CN117357632A CN 117357632 A CN117357632 A CN 117357632A CN 202311436301 A CN202311436301 A CN 202311436301A CN 117357632 A CN117357632 A CN 117357632A
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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Abstract
The invention relates to the technical field of biological medicines, in particular to application of IL-33 recombinant protein in preparing a medicine for treating acute liver failure, relieving liver immune injury or relieving liver inflammatory injury. Experiments prove that the IL-33 recombinant protein can obviously promote the expression of PD-1 and Ki67 in spleen Treg cells and promote the expression of CTLA4 in liver Treg cells, thereby enhancing the inhibition function of the Treg cells and relieving liver immune injury. Therefore, the IL-33 recombinant protein can be used for preparing a novel medicament for treating acute liver failure or relieving liver immune injury. The invention can effectively provide a new molecular target for treating acute liver failure, relieving liver immune injury or relieving liver inflammatory injury, and provide guidance for drug development or formulation of new treatment schemes.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of IL-33 recombinant protein in preparing a medicine for treating acute liver failure, relieving liver immune injury or relieving liver inflammatory injury.
Background
In liver failure, a great deal of liver cells die, inflammatory cell infiltration and liver ischemic injury are accompanied, and the regeneration capacity of the liver cells is exceeded, so that liver function is decompensated, a series of complications are caused, and finally, the multi-organ failure is caused, and the mechanism mainly relates to direct injury and immune-mediated indirect injury. Direct damage mainly refers to the direct toxic effects of viruses, drugs, poisons, alcohol and other factors on the liver. However, immune mediated indirect injury is critical in the course of liver failure, especially the activation of T cell immune cells, and the imbalance of immune cells, which produce corresponding inflammatory cytokines, causing inflammatory waterfall-like responses. Therefore, how to regulate unbalanced immune cells, block inflammatory cytokine storm and promote regeneration and repair of liver cells is an important and difficult problem of acute liver failure research.
Interleukin 33 (IL-33) is a multifunctional gene found in 2005, is one of important regulatory factors for inflammatory response and immune bias, and can act as a transcription factor for molecules located in the nucleus and can be secreted outside cells to act as a cytokine. IL-33 is a novel member of the IL-1 family, one of the important mediators of inflammatory response and immune bias, and the receptor is ST2 (Interleukin 1like receptor), mainly inducing Th2 type immunity. IL-33 normally exists in the nucleus and functions as a transcription factor, and when the cell is damaged, IL-33 is secreted to the outside and then binds to the receptor ST2, thereby functioning as a cytokine. However, no report on the use of IL-33 recombinant proteins for treating liver failure or reducing liver immune injury is currently seen.
Disclosure of Invention
First, the technical problem to be solved
In view of the above-mentioned shortcomings and disadvantages of the prior art, the present invention provides an IL-33 recombinant protein, which can be used to increase proliferation activity of Treg cells in liver, promote expression of PD-1 and the like in spleen Treg cells, promote expression of CTLA4 and the like in liver Treg cells to enhance inhibition function of Treg cells, thereby reducing immune injury degree of liver and achieving the purpose of treating acute liver failure.
(II) technical scheme
The technical scheme of the invention is as follows:
the application of IL-33 recombinant protein in preparing medicaments for treating acute liver failure, relieving liver immune injury or relieving liver inflammatory injury.
Preferably, the liver immune injury is an immune system dysfunction, resulting in liver injury; including autoimmune factors, genetic factors, viral factors, drug or alcohol induced immune injury of the liver.
Preferably, the liver immune injury is a Con a-induced liver injury.
Preferably, the amino acid sequence of the IL-33 recombinant protein is shown as SEQ ID NO. 1.
Preferably, the medicament is an injection containing IL-33 recombinant protein.
(III) beneficial effects
Experiments prove that the change trend of IL-33 in serum of Con A liver injury model animals is consistent with the liver tissue injury degree. However, if the IL-33 recombinant protein is used for pretreatment before the modeling of the mouse Con a, the inflammatory injury of the liver of the mouse induced by the Con a (canavalin) can be significantly reduced, and the specific manifestations include: can obviously reduce the liver ALT and AST levels of a Con A liver injury model mouse, lighten the liver lobular structural disorder of the mouse and obviously reduce the liver necrosis area, and simultaneously obviously lighten inflammatory cell infiltration. Experiments further prove that after Con A modeling is carried out on mice pretreated by IL-33 recombinant protein, the spleen Treg cells and the control group have no obvious difference, but the liver Treg cells are obviously increased, and the IL-33 recombinant protein can obviously promote the expression of PD-1 and Ki67 in the spleen Treg cells and promote the expression of CTLA4 in the liver Treg cells, so that the inhibition function of the Treg cells is enhanced, and the liver immune injury degree is reduced. Therefore, the IL-33 recombinant protein can be used for preparing a novel medicament for treating acute liver failure or relieving liver immune injury.
The invention can effectively provide a new molecular target for the treatment of acute liver failure and theoretical guidance for drug development or formulation of new treatment schemes.
Drawings
FIG. 1 shows changes in liver injury in mice before and after Con A modeling; a is the ALT change of plasma of a WT mouse 6, 12 and 24 hours before (control) and after Con A molding; b is Con A before molding (control) and Con A after molding 6, 12 and 24 hours, WT mouse plasma AST changes; c is a graph represented by H & E staining of liver tissue of a WT mouse 6, 12 and 24 hours before (represented by 0) and after Con A molding; d is the Ishak inflammatory injury score of HE staining of liver tissue of 6, 12, 24h before (control) and after Con a molding.
FIG. 2 shows IL-33 expression in serum of mice before Con A modeling (control); a is ELISA method to detect IL-33 expression level (pg/mL) of WT mouse plasma before Con A modeling and after Con A modeling at 6, 12 and 24 hours; b is the relative expression level of IL-33mRNA in liver tissues of WT mice 6, 12 and 24 hours before (control) Con A molding and after Con A molding detected by qRT-PCR method.
FIG. 3 is a graph showing the effect of IL-33 recombinant protein on liver injury; a is the difference of plasma ALT level of Con A before molding (control), molding for 12h (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein); b is the difference of plasma AST level of Con A before molding (control), molding for 12h (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein); c is HE staining results of liver tissues of a Con A pre-molding (control), molding 12h (Con A group) and an IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein); d is the pathological score of liver injury in Con A pre-molding (control), molding 12h (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein).
FIG. 4 is the effect of IL-33 recombinant protein on spleen and liver Treg cells; A. e is the proportion of Treg cells in spleen and liver of mice in Con A pre-molding (control), 12h molding (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein); B. f is PD-1 in spleen and liver Treg cells of mice of Con A pre-molding (control), molding 12h (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) + Is a ratio of cells; c, G is the cell proportion of CTLA4+ in spleen and liver Treg cells of mice of Con A before molding (control), 12h (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein); d, H is Ki67 in spleen and liver Treg cells of mice of Con A pre-molding (control), molding 12H (Con A group) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) + Is a cell proportion of (a).
Detailed Description
The invention will be better explained by the following detailed description of the embodiments with reference to the drawings.
Numerous studies have found that the serum and liver of patients with acute liver injury have significantly increased helper T cells 17 (Th 17) cells, and that the pro-inflammatory cytokines IL-17A, IL-22, CXCL8, and GM-CSF, etc., have increased, while Regulatory T cells (Tregs) have decreased inhibitory function. Thus, if the inhibitory function of regulatory T cells could be promoted or enhanced, it would be helpful to alleviate hepatic immune injury. According to animal experiments, the IL-33 recombinant protein can obviously promote the expression of PD-1 and Ki67 in spleen Treg cells and promote the expression of CTLA4 in liver Treg cells, so that the inhibition function of the Treg cells is enhanced, the immune injury degree of the liver is reduced, and acute liver failure is treated. Therefore, the IL-33 recombinant protein can be used for preparing medicines for treating acute liver failure, relieving liver immune injury or promoting regeneration and repair of liver cells.
The invention is illustrated below in connection with specific examples and experimental data/graphs. In the following examples, when comparing liver or spleen changes in mice at various treatment groups or different time points, serum is collected to detect liver function indexes ALT and AST, liver tissue HE is stained to analyze pathological damage, and the liver tissue HE is scored according to Ishak modified liver inflammatory injury scoring criteria (mainly comprising 4 aspects of periportal or outer peripheral surface inflammation, fusion necrosis, focal necrosis and manifold area inflammation, the scoring follows double-blind principle), and expression level of mouse plasma IL-33 is detected by ELISA, and liver mesenchymal cells and spleen cell changes are detected by flow cytometry. The detection method or sample processing method according to each embodiment is as follows:
1. mouse serum liver function index ALT and AST detection:
(1) after sacrificing the mice, collecting mouse plasma, and freezing at-80 ℃ or directly detecting; (2) diluting the plasma 10 times with physiological saline before detection; (3) mouse plasma levels of glutamic pyruvic transaminase (Alanine transaminase, ALT) and glutamic oxaloacetic transaminase (Aspertate aminotransferase, AST) were measured using a fully automatic biochemical analyzer of Japan Olympus-AU 2700.
2. Liver tissue HE staining:
(1) placing the tissue in an embedding box, and washing for multiple times to clean the surface of the tissue from paraformaldehyde;
(2) dehydrating the tissue block, embedding the tissue block in paraffin, slicing the tissue block to a slice thickness of 4 mu m, paving the slice on a glass slide, and baking the slice at 75 ℃ for 2 hours;
(3) dewaxing: dewaxing with xylene for 5 times, each time greater than 30min;
(4) hydration: subjecting the tissue slice to double-distilled dewaxing sequentially with 2 times of absolute methanol, 2 times of 95% alcohol, 1 time of 85% alcohol and 2 times of 75% alcohol;
(5) staining with hematoxylin for 10min, and washing with clear water;
(6) dyeing with eosin for 5min, and washing with clear water;
(7) differentiating with 1% hydrochloric acid alcohol for 30s, and turning blue with ammonia water for 3min;
(8) dehydrating and sealing the sheet: the tissue slice is dehydrated and transparent by 75 percent alcohol for 2 times, 85 percent alcohol for 1 time, 95 percent alcohol for 2 times, absolute alcohol for 2 times and xylene for 2 times in sequence, and then is sealed by neutral resin;
(9) microscopic observations were made using 100×,200×, 400× magnification and scoring was performed according to the Ishak modified version of liver inflammatory injury scoring criteria (see table 1), consisting essentially of 4 aspects of periportal or outer peripheral surface inflammation, fusion necrosis, focal necrosis, and inflammation of the ductal region, following the double blind principle.
Table 1: ishak modified version liver inflammatory injury score (0-18 points)
3. ELISA detection of mouse plasma IL-33 expression levels:
(1) collecting plasma of mice before Con A molding (control), after Con A molding for 12h, and after Con A molding and IL-33 pretreatment group mice, and collecting blood by eyeball blood collection;
(2) preparing ELISA reagent standard according to the specification;
(3) adding the sample and standard substances (100 mu L/hole) with different concentrations into ELISA plate holes (blank control group is added with sample diluent), sealing the reaction holes by using sealing plate gummed paper, and incubating for 90 minutes at 37 ℃;
(4) washing the plate for 4 times: throwing out liquid in the holes, adding 350 mu L of washing liquid into each hole, standing for 30s, throwing out liquid, and beating on thick water-absorbing paper;
(5) biotinylated antibody working solution (100. Mu.L/well) was added. Sealing the reaction hole by using sealing plate gummed paper, and incubating for 60 minutes at 37 ℃;
(6) washing the plate for 4 times;
(7) enzyme conjugate working solution (100. Mu.L/well) was added. Sealing the reaction hole by using sealing plate gummed paper, and incubating for 30 minutes at 37 ℃;
(8) washing the plate for 4 times;
(9) adding 100 mu L/hole of a color reagent, and incubating for 10-20 minutes at 37 ℃ in dark place;
to 100. Mu.L/well of stop solution, and the OD450 value (within 5 minutes) was measured immediately after mixing.
4. qRT-PCR method for detecting liver IL-33 expression:
(1) the tissue was weighed 20-30mg into a 1.5mL centrifuge tube, 350. Mu.L of RLT lysate was added and ground using a freeze grinder. The ground mixture was centrifuged at 13000 Xg for 3min and the whole supernatant was transferred to a purge column (purge column placed in collection tube);
(2) accurately measuring the volume of the filtered liquid by using a pipetting gun, adding 70% ethanol with equal volume, and immediately blowing and uniformly mixing;
(3) immediately transferring the mixture to an adsorption column (the adsorption column is placed in a collection tube), centrifuging for 30s at 13000 Xg, and discarding the filtrate;
(4) adding 700 μl of the rinse solution 1, standing at room temperature for 1min, centrifuging at 13000×g for 30s, and discarding the filtrate;
(5) adding 500 μl of rinse solution 2, centrifuging at 12000×g for 30s, and discarding the filtrate;
(6) repeating the previous step;
(7) placing the adsorption column back into a collecting pipe, and centrifuging with 13000 Xg for 2min to spin-dry the adsorption column to thoroughly remove residual ethanol;
(8) the column was placed in a 1.5mL RNase-Free centrifuge tube, 30. Mu.L-50. Mu.L of the eluent was added to the center of the column membrane, and the column was allowed to stand for 1min and centrifuged at 12000 Xg for 1min.
(9) The collected RNA is put into a refrigerator at-80 ℃ for freezing or directly subjected to downstream experiments.
Placing the extracted RNA on ice to defrost, and preparing a reverse transcription system according to the formula of the table 2; reverse transcription was performed according to the procedure of Table 3 to obtain cDNA;
table 2: reverse transcription reaction system
Table 3: reverse transcription procedure
Diluting the cDNA after reverse transcription by 10 times with sterile and asepsis water;
primers required for qRT-PCR were synthesized according to Table 4;
table 4: qRT-PCR primer sequences
Preparing a qRT-PCR system according to the scheme of Table 5, and performing qRT-PCR according to the procedure of Table 6;
qRT-PCR results were normalized to control, and the calculation method followed 2-DeltaCT.
Table 5: qRT-PCR system
Table 6: qRT-PCR reaction procedure
5. Preparation of liver mesenchymal cell suspension
(1) After sacrificing the mouse, exposing the abdomen of the mouse, finding out portal veins and inferior vena cava, injecting normal saline through portal vein puncture, cutting off inferior vena cava, and flushing blood stasis in the liver;
(2) mouse livers were taken in 10cm dishes containing 8mL of 1 XPEB, and grinded using a cell filter (cell filter) with a filter membrane pore size of 70 μm, and the suspension was collected in a 15mL centrifuge tube;
(3) centrifuging at 2000rpm at 4deg.C for 10min, discarding supernatant, and re-suspending;
(4) re-mixing the cells in 8mL of 40% percoll, lightly paving the cells on the upper layer of a centrifuge tube containing 4mL of 80% percoll, centrifuging at 2000rpm and 4 ℃ for 20min, and adjusting the lifting speed of the centrifuge to 9 and 4;
(5) after centrifugation, a layer of cloud-like cell layer is arranged between 40% of percoll and 80% of percoll, and the cell layer is carefully sucked into a 15mL centrifuge tube containing 4mL of 1 XPBEB and uniformly mixed;
(6) centrifuging at 1500rpm at 4deg.C for 10min, discarding supernatant, and re-suspending;
(7) adding 2mL of 1 XPEB, mixing, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending;
(8) adding 2mL of 1 Xerythrocyte lysate, and performing room temperature lysis for 10min;
(9) adding 1mL of 1 XPEB to terminate the lysis, mixing, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending;
adding 2mL of 1 XPEB, uniformly mixing, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and uniformly re-suspending;
staining was performed by flow cytometry.
6. Preparation of spleen Single cell suspension
(1) After sacrificing the mice, taking spleens of the mice into a culture dish with the diameter of 6cm and containing 4mL of PBEB, roughly grinding the spleens by using a glass slide, and sucking the grinded spleens suspension into a flow type tube;
(2) centrifuging at 1200rpm at 4deg.C for 5min, discarding supernatant, and re-suspending;
(3) split red: adding 2mL of erythrocyte lysate into each tube, mixing uniformly by vortex, and cracking for 10min at room temperature;
(4) terminating the cleavage; adding 1mL of PBEB into each tube to terminate the cracking, vortex and mix uniformly, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending uniformly;
(5) and (3) filtering: adding 4mL of PBEB into each tube, mixing by vortex, filtering the cell suspension into another flow tube by a 400-mesh filter membrane, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and mixing by re-suspension;
(6) 4mL of PBEB is added to each tube, and vortex mixing is carried out for subsequent experiments.
7. Flow cytometry staining
(1) The obtained liver and spleen single cell suspension is divided into new flow tubes according to 10-7 cells;
(2) adding anti-CD 3, anti-CD 4, anti-CD 25 and anti-PD-1 antibodies into each tube of cells according to the antibody specification, mixing uniformly by vortex, and keeping out of light for 15min at 4 ℃;
(3) adding 2mL of PBEB into each tube, mixing by vortex, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending;
(4) rupture of membranes: adding 500 mu L of fixed rupture liquid into each tube, mixing uniformly by vortex, and keeping out of light for 40min at 4 ℃;
(5) adding 1mL of membrane rupture cleaning liquid into each tube, mixing by vortex, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending;
(6) adding PBEB to a volume of 150 mu L into each tube, adding anti-Foxp 3, anti-CTLA 4 antibody and anti-Ki-67 antibody according to an antibody specification, and mixing by vortex at 4 ℃ for 30min in a dark place;
(7) adding 2mL of PBEB into each tube, mixing by vortex, centrifuging at 1200rpm and 4 ℃ for 5min, discarding the supernatant, and re-suspending;
(8) flow cytometry detection was performed.
Example 1
This example demonstrates that in a Con A (Canavalia) induced mouse model, IL-33 expression is consistent with a trend in the severity of liver injury. In order to compare the change rule of the liver injury of the mice in the Con A induced liver injury mouse model, mice are sacrificed 6 hours, 12 hours and 24 hours after the Con A molding, and the liver transaminase and the liver pathological injury conditions of the mice at different time points are compared. The molding method of the embodiment comprises the following steps: female C57BL/6J mice were randomly selected from 6-8 weeks and divided into two groups, namely a Con A model group and a control group, and the treatment method is as follows:con A model group: administration of 10mg/kg of Canavalia gladiata protein was injected via the inner canthus vein;control group: the physiological saline is administered in a corresponding dose.
The experimental results are shown in FIG. 1. In the figure, A is: pre-Con a molding (control group) and post-Con a molding 6, 12, 24h, wt mouse plasma ALT changes (n=5-6); b is: pre-Con a molding (control group) and post-Con a molding 6, 12, 24h, wt mouse plasma AST changes (n=5-6); c is: con A pre-molding (control) and Con A post-molding 6, 12, 24H, WT mouse liver tissue H & E staining represent plots (100×,200×). D is: pretreatment of Con A (control) and 6, 12, 24h after Con A molding, liver tissue HE staining Ishak inflammatory injury score (n=6). ns represents no statistical differences, without statistical differences. * And P <0.05, P <0.01, P <0.001, respectively.
From the results shown in FIG. 1, the ALT and AST were significantly increased after Con A molding, and reached a peak 12 hours after molding, and then gradually decreased (A, B in FIG. 1) as compared with the control group. HE staining of liver tissue showed: after Con A molding, liver lobular structure was disturbed, inflammatory cell infiltration in the sink region was remarkable, the necrotic area of liver tissue was enlarged, and the peak was reached at 12 hours, bridging necrosis occurred (C in FIG. 1), and after Con A molding for 24 hours, the necrotic area of liver in mice was remarkably reduced (C in FIG. 1). Ishak liver tissue inflammatory injury scores also showed a significant increase in liver injury scores after Con A induction, a peak 12 hours and a significant decrease in 24 hours (D of FIG. 1). The results suggest that the liver injury of mice peaks 12 hours after Con A molding.
Meanwhile, the expression of IL-33 in Con A mouse model was further examined in the experiment, and the results are shown in FIG. 2. In the figure, A is: ELISA method for detecting IL-33 expression level (n=8-11) of WT mice plasma 6, 12 and 24 hours before Con A molding (control) and after Con A molding; b is: qRT-PCR method detects IL-33mRNA relative expression level (n=4-6) in liver tissue of WT mouse 6, 12 and 24 hours before Con A molding (control) and after Con A molding; ns represents no statistical differences, without statistical differences. * P <0.01, P <0.001, respectively.
As can be seen from the results shown in FIG. 2, the IL-33 expression was gradually increased after Con A molding, reached a peak (163.2.+ -. 58.83 pg/mL) after 12 hours, and decreased (36.90.+ -. 40.47 pg/mL) after 24 hours, compared with the control group (5.167.+ -. 4.036 pg/mL) (A in FIG. 2). The detection of IL-33mRNA transcription level of liver tissue shows that after Con A modeling, the IL-33 transcription level in liver tissue is gradually increased, the transcription level reaches a peak for 12 hours, the transcription level is about 18 times that of a control group, and the transcription level is about 2 times that of the control group after 24 hours (B of FIG. 2). Taken together with fig. 1 and 2, the results suggest that IL-33 expression in serum of model mice after Con a modeling gradually increases, peaks at 12h, and then gradually decreases, and the change trend is consistent with the degree of liver tissue damage.
Example 2
This example demonstrates that intervention with IL-33 recombinant protein significantly reduces the extent of Con A-induced inflammatory injury to the liver of mice. In order to clearly demonstrate the role of IL-33 recombinant protein in liver inflammatory injury, the present experiment pre-treats mice by injecting IL-33 recombinant protein before Con A molding, then normally molds Con A, compares the liver injury difference of two molded mice which are not treated by IL-33 recombinant protein and are treated by IL-33 recombinant protein, and the comparison indexes comprise: serum detection liver function indexes ALT and AST, liver tissue HE staining analysis pathological injury condition, ishak modified liver inflammatory injury score, plasma IL-33 expression level, liver IL-33mRNA expression level, mouse liver and spleen Treg functions and the like (comprising cell proliferation PD-1 and CTLA-4 expression). By detecting the above indicators, the effect of IL-33 on Treg function was comprehensively evaluated.
The molding method of the embodiment comprises the following steps: female C57BL/6J mice were randomly selected from 6-8 weeks and divided into three groups, namely IL-33 recombinant protein (pretreatment) group, con A group and control group, and the operations were as follows:
con A model group: 2 hours before Con A molding, physiological saline was injected intraperitoneally into 1 g/mouse, and 10mg/kg (Canavalia) of the saline was then administered via intracanthus intravenous injection to perform Con A molding.IL-33 recombinant protein (pretreatment) group: the molding was performed by intraperitoneal injection (IL-33 recombinant protein) in 1 g/mouse 2 hours before Con A molding, followed by injection of 10mg/kg of Canavalia gladiata protein via the inner canthus vein.Control group: the corresponding dose of physiological saline is always administered.
Wherein, the amino acid sequence of the IL-33 recombinant protein used for injection of the IL-33 recombinant protein group is SIQGTSLLTQSPASLSTYNDQSVSFVLENGCYVINVDDSGKDQEQDQVLLRYYESPCPASQSGDGVDGKKLMVNMSPIKDTDIWLHANDKDYSVELQRGDVSPPEQAFFVLHKKSSDFVSFECKNLPGTYIGVKDNQLALVEEKDESCNNIMFKLSKI (see SEQ ID NO. 1).
The experimental results are shown in FIG. 3. In the figure, A is: differences in plasma ALT levels (n=6) in Con a pre-modeling (control), 12h (Con a), and IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein); in the figure, B is: plasma AST level differences (n=6) in the Con a pre-model (control), model 12h (Con a) and IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein). C is: liver tissue HE staining results (100×,200×) of the Con a pre-molding (control), molding 12h (Con a) and IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein). D is: pre-modeling (control), 12h (Con a), and IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein) liver injury pathology score (n=5). * And P <0.05, P <0.01, P <0.001, respectively.
As can be seen from FIG. 3, the liver ALT and AST levels of mice were significantly increased (ALT: 4897.+ -. 1578U/L, AST: 5468.+ -. 1923U/L) after 12h of Con A molding compared with the control group (ALT: 41.33.+ -. 15.95U/L, AST: 131.5.+ -. 37.82U/L), while the IL-33 recombinant protein (pretreatment) levels of mice were significantly decreased (ALT: 1522.+ -. 1377U/L, AST: 1766.+ -. 1314U/L) (A, B of FIG. 3). Liver HE staining showed that the IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein) mice had a reduced liver lobular structural disturbance, a significantly reduced liver necrosis area and significantly reduced inflammatory cell infiltration compared to Con a model 12h (C of fig. 3). Meanwhile, ishak liver tissue pathology injury scores also showed that the IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) had significantly lower liver injury scores compared to the Con A model (D of FIG. 3).
Example 3
Based on the experimental method and model induction method of example 2, this example further compares the changes in mouse Treg cells and Treg cell functional molecules of the IL-33 recombinant protein pretreatment group under the Con a model. The experimental results are shown in fig. 4, in which a and E are respectively: pre-modeling (control), modeling for 12h (Con a), and ratio of Treg cells in spleen and liver of IL-33 recombinant protein pretreatment group (Con a+il-33 recombinant protein) mice (n=7); in the figure, B and F are respectively: PD-1 in mice spleen and liver Treg cells of Con A Pre-model (control), model 12h (Con A) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) + Is a ratio of cells; in the figure, C, G are respectively: CTLA4 in mice spleen and liver Treg cells of Con A pre-model (control), model 12h (Con A) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) + Is a ratio of cells; in the figure, D, H are respectively: ki67 in mice spleen and liver Treg cells of Con A pre-model (control), model 12h (Con A) and IL-33 recombinant protein pretreatment group (Con A+IL-33 recombinant protein) + Is a cell proportion of (a). ns represents no statistical differences, without statistical differences. * Respectively representing P<0.05，P<0.01，P<0.001。
From the results shown in fig. 4, after modeling of Con a, no significant difference was seen between Treg cells in spleen and control (as in fig. 4 a), while Treg cells in liver were significantly increased (as in fig. 4E). As seen in a and E of fig. 4, after pretreatment with IL-33 recombinant protein, although there was no significant difference between Treg cells and the Con a model group, it was not difficult to find from B and D of fig. 4 that IL-33 recombinant protein significantly promoted the expression levels of PD-1, ki67 in spleen Treg cells, while significantly promoting the expression level of CTLA4 in liver Treg cells (e.g., G of fig. 4), but after pretreatment with IL-33 recombinant protein, there was no significant effect on CTLA4 expression in spleen Treg cells and PD-1 and Ki67 expression in liver Treg cells (C, F, H of fig. 4), i.e., neither promotion nor inhibition. Thus, the proliferation activity of the liver and spleen Treg cells can be improved by injecting the IL-33 recombinant protein, and the inhibition function of the Treg cells can be enhanced by promoting the expression of spleen PD-1 and Ki67 and the expression of liver CTLA-4, so that the acute liver failure injury is relieved.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (5)
- Application of IL-33 recombinant protein in preparing medicine for treating acute liver failure, relieving liver immune injury or relieving liver inflammatory injury.
- 2. The use according to claim 1, wherein the liver immune injury is an immune system dysfunction resulting in liver injury; including autoimmune factors, genetic factors, viral factors, drug or alcohol induced immune injury of the liver.
- 3. The use according to claim 3, wherein the liver immune injury is a Con a-induced liver injury.
- 4. The use according to claim 1, wherein the amino acid sequence of the IL-33 recombinant protein is shown in SEQ ID No. 1.
- 5. The use according to claim 1, wherein the medicament is an injection comprising an IL-33 recombinant protein.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111544574A (en) * | 2020-06-16 | 2020-08-18 | 天津大学 | Application of interleukin 5 in preparation of medicine for preventing, treating or relieving liver injury |
| CN112210556A (en) * | 2020-10-15 | 2021-01-12 | 扬州大学 | Group of shRNA for targeted interference of IL-33 expression, recombinant adenovirus vector, and construction method and application thereof |
| CN113588964A (en) * | 2021-07-08 | 2021-11-02 | 苏州市第五人民医院 | Slow-acceleration acute hepatic failure detection kit and preparation method thereof |
| CN114555109A (en) * | 2019-10-11 | 2022-05-27 | 交晨生物医药技术(上海)有限公司 | Methods of treating cancer using IL-33 proteins |
-
2023
- 2023-11-01 CN CN202311436301.4A patent/CN117357632A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114555109A (en) * | 2019-10-11 | 2022-05-27 | 交晨生物医药技术(上海)有限公司 | Methods of treating cancer using IL-33 proteins |
| CN111544574A (en) * | 2020-06-16 | 2020-08-18 | 天津大学 | Application of interleukin 5 in preparation of medicine for preventing, treating or relieving liver injury |
| CN112210556A (en) * | 2020-10-15 | 2021-01-12 | 扬州大学 | Group of shRNA for targeted interference of IL-33 expression, recombinant adenovirus vector, and construction method and application thereof |
| CN113588964A (en) * | 2021-07-08 | 2021-11-02 | 苏州市第五人民医院 | Slow-acceleration acute hepatic failure detection kit and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| HILAL AHMAD KHAN 等: "IL-33 ameliorates liver injury and inflammation in Poly I:C and Concanavalin-A induced acute hepatitis", MICROBIAL PATHOGENESIS, vol. 150, 29 December 2020 (2020-12-29), pages 3 - 5, XP086473297, DOI: 10.1016/j.micpath.2020.104716 * |
| MOK MY 等: "ACCESSION No. NP_001158196.1 interleukin-33 isoform 1 [Mus musculus]", GENBANK数据库, 24 September 2023 (2023-09-24) * |
| 姜绍文 等: "IL-33及其受体ST2在D-GalN/LPS诱导的急性肝功能衰竭小鼠中的表达及意义", 肝脏, vol. 21, no. 4, 30 April 2016 (2016-04-30), pages 267 - 272 * |
| 郑晨宏;谢晓华;: "刀豆蛋白A诱导肝损伤模型中免疫细胞及因子研究进展", 解放军医学院学报, vol. 37, no. 2, 3 November 2015 (2015-11-03), pages 193 * |
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