Application of deoxyribonuclease I in preparation of medicine for treating wrapped peritoneal sclerosis
By using DNase I to inhibit the formation of macrophage extracellular traps, the problem of lack of effective treatments for encapsulated peritoneal sclerosis is solved, abdominal adhesions and inflammation are significantly improved, and a new prevention and treatment strategy for encapsulated peritoneal sclerosis is provided.
Patent Information
- Application Number
- CN202411393209.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-08
- Publication Date
- 2025-01-21
AI Technical Summary
The prior art lacks effective treatments to deal with the severe peritoneal dialysis complications of encapsulated peritoneal sclerosis (EPS), resulting in poor prognosis and high mortality rates in patients.
Deoxyribonuclease I (DNase I) is used as an inhibitor of macrophage extracellular traps (METs). DNase I solution is injected intraperitoneally to inhibit the formation of METs and reduce abdominal adhesion and inflammation.
DNase I significantly reduced the expression of METs in the peritoneal tissue of mice with encapsulated peritoneal sclerosis, improved the abdominal adhesion score, peritoneal inflammatory infiltration, collagen deposition area in the adhesion area, and peritoneal thickness, thereby improving related pathological changes.
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Abstract
Description
Technical Field
[0001] The present application relates to the field of biomedicine technology, and in particular to the application of deoxyribonuclease I in the preparation of a drug for treating encapsulated peritoneal sclerosis. Background Art
[0002] Encapsulating peritoneal sclerosis (EPS) is one of the most serious complications of peritoneal dialysis. It is also called abdominal cocooning or sclerosing encapsulating peritonitis. Its main characteristics are intermittent or persistent gastrointestinal obstruction symptoms, and peritoneal thickening, hardening, calcification or fiber encapsulating the small intestine confirmed by macroscopic and / or imaging. Encapsulating peritoneal sclerosis has a poor prognosis and a high mortality rate. However, the mechanism of the occurrence and development of EPS abdominal adhesions is still poorly understood, and there is a lack of effective treatment methods.
[0003] Extracellular traps (ETs) are a new programmed cell death pathway, called "ETosis", and in neutrophils, they are called "NETosis", that is, neutrophile extracellular traps (NETs). When cells are stimulated by multiple external factors, protein arginine deiminase type-4 (PAD4) in their nuclei is activated, mediating the arginine on histones to undergo citrullination, leading to chromatin depolymerization and cytoskeleton disintegration. Subsequently, nuclear contents including chromatin are mixed with proteins in the cytoplasm such as myeloperoxidase (MPO) and neutrophile elastase (NE), and are squeezed out of the cell body to form a mesh structure outside the cell that can capture and kill microorganisms. The basic skeleton is composed of DNA fibers with a diameter of 15-17nm and a spherical region, covered with citrullinated histone H3, MPO, NE and other protein particles. In addition to neutrophils, other immune cells such as macrophages, monocytes, mast cells, eosinophils and basophils can also produce "extracellular trapping net" structures.
[0004] ETosis was originally described as an antibacterial protective mechanism for capturing and killing bacteria. As research on ETosis deepens, it has been found that ETs are also produced in a variety of sterile inflammatory diseases and have pathogenic effects. Stimuli that induce the formation of ETs in vivo include but are not limited to microorganisms and their products, inflammatory cytokines and chemokines, immune complexes, autoantibodies, crystals, chemicals, and activated platelets. Excessive production or impaired degradation of NETs can lead to the initiation or development of a variety of diseases.
[0005] Currently, there is no drug that can effectively treat EPS complications caused by long-term peritoneal dialysis, peritonitis, abdominal surgery, etc., thereby improving the prognosis of the above patients. Summary of the invention
[0006] The purpose of the present application is to overcome the deficiencies of the above-mentioned prior art and provide a method for using deoxyribonuclease I in the preparation of a drug for treating encapsulating peritoneal sclerosis.
[0007] To achieve the above purpose, the technical solution adopted by this application is:
[0008] The present application provides the use of deoxyribonuclease I in preparing an inhibitor of macrophage extracellular traps.
[0009] The inventor of the present application unexpectedly discovered through a large number of experiments that deoxyribonuclease I can be used as an inhibitor of macrophage extracellular traps. Deoxyribonuclease I (DNase I) is a naturally occurring enzyme in the body, and has the characteristics of high safety and low toxicity. At the same time, because the peritoneum is in a state of aseptic inflammation under conventional peritoneal dialysis and EPS conditions, the use of deoxyribonuclease I to ablate the existing extracellular trap structure in the peritoneal cavity will not cause peritoneal infection, providing a better choice for peritoneal immunotherapy, which can prevent or improve EPS peritoneal adhesions while avoiding the risk of infection and hepato-renal toxicity as much as possible.
[0010] The present application also provides the use of deoxyribonuclease I as an extracellular trap inhibitor in the preparation of a drug for treating encapsulating peritoneal sclerosis.
[0011] This application uses scRNA-seq analysis of peritoneal dialysis effluent cells to find that there is a significant fibrosis process in the peritoneal environment of EPS patients, and it is related to the effect of macrophages on fibroblasts. Subsequent macrophage subpopulation classification and enrichment analysis found that pro-inflammatory macrophages (S100A8 + The proportion of macrophages in the mitochondria was significantly increased, and the macrophage extracellular trap (METs) formation pathway was significantly enriched in these cells.
[0012] This application constructed an EPS mouse model by intraperitoneal injection of SHS solution (Surgical hygiene solution) and found that fibroblasts and macrophages are the main cellular components in the peritoneal adhesion area, and the expression level of METs is positively correlated with the severity of EPS.
[0013] The present application unexpectedly discovered that the use of deoxyribonuclease I can inhibit METs. Deoxyribonuclease I can effectively reduce the expression of METs in the peritoneal tissue of mice with encapsulating peritoneal sclerosis, and can also significantly improve the abdominal adhesion score, peritoneal inflammatory infiltration, collagen deposition area in the adhesion area, and peritoneal thickness of mice with encapsulating peritoneal sclerosis, thereby improving the pathological changes associated with encapsulating peritoneal sclerosis. Therefore, the present application constructs a new strategy for the prevention and treatment of encapsulating peritoneal sclerosis by targeting METs to treat encapsulating peritoneal sclerosis.
[0014] As a preferred embodiment of the application described in the present application, the extracellular trap inhibitor is a macrophage extracellular trap inhibitor.
[0015] According to animal experiments, the use of the deoxyribonuclease I of the present application can significantly inhibit the production of METs on the peritoneal surface.
[0016] As a preferred embodiment of the application described in the present application, the deoxyribonuclease I is prepared into a deoxyribonuclease I solution using deionized water as a solvent.
[0017] The solvent may also be a buffer commonly used in the art, and is not limited to deionized water.
[0018] As a preferred embodiment of the application described in the present application, the concentration of deoxyribonuclease I in the deoxyribonuclease I solution is 5 mg / mL.
[0019] As a preferred embodiment of the application described in the present application, the deoxyribonuclease I solution is stored at a temperature of -20°C to -30°C for future use, and the deoxyribonuclease I solution is injected intraperitoneally.
[0020] As a preferred embodiment of the application described in the present application, the effective dosage of the deoxyribonuclease I used in animal experiments is 5 mg / kg.
[0021] As a preferred embodiment of the application described in the present application, the encapsulating peritoneal sclerosis includes the encapsulating peritoneal sclerosis formed as a result of peritoneal dialysis or abdominal surgery.
[0022] Deoxyribonuclease I treatment significantly improved the mutual adhesions between the abdominal organs and between the abdominal organs and the parietal peritoneum of mice, alleviated the symptoms of intestinal obstruction such as slowed intestinal peristalsis and enlarged intestinal cavity, and reduced the adhesion score and IL-6 level in the peritoneal lavage fluid, indicating that ablation of METs structure by deoxyribonuclease I reduced the level of peritoneal inflammation of encapsulating peritoneal sclerosis and improved abdominal adhesions.
[0023] In addition, the results of histopathological examination also showed that MASSON staining of the mouse abdominal cross-section showed that the degree of adhesion of the abdominal organs in the deoxyribonuclease I treatment group was significantly improved compared with the Saline group, and the collagen deposition in the adhesion area was significantly reduced; deoxyribonuclease I treatment also significantly reduced the thickness of the parietal peritoneum and visceral peritoneum of EPS mice, as well as the deposition of ECM-related proteins such as α-SMA, COL1A1, and Fibronectin on the peritoneal surface. The above results suggest that ablating the METs that have already been produced in the abdominal cavity through deoxyribonuclease I treatment can effectively alleviate peritoneal fibrosis and ECM deposition on the peritoneal surface, and ultimately improve the formation of intra-abdominal organ adhesions.
[0024] As a preferred embodiment of the application described in the present application, the deoxyribonuclease I specifically inhibits the extracellular traps of macrophages, thereby treating encapsulating peritoneal sclerosis.
[0025] As a preferred embodiment of the application described in this application, the application includes at least one of the following aspects:
[0026] 1) DNase I significantly reduced the expression of METs in the peritoneal cavity;
[0027] 2) Deoxyribonuclease I improves peritoneal adhesion and inflammation in mice with encapsulated peritoneal sclerosis;
[0028] 3) Deoxyribonuclease I improves peritoneal thickening and fibrosis in mice with encapsulating peritoneal sclerosis.
[0029] As a preferred embodiment of the application described in the present application, the drug also includes excipients commonly used in pharmacy.
[0030] At present, the research focusing on the extracellular trap of macrophages requires the construction of mice with conditional knockout of the PAD4 gene in macrophages (Pad fl / fl The construction and breeding of conditional knockout mice requires a long time and a lot of money. In vivo experiments have shown that mice with partial knockout of the PAD4 gene cannot specifically knock out the extracellular traps of macrophages, and therefore cannot completely rule out the influence of neutrophils.
[0031] In patients with encapsulating peritoneal sclerosis (EPS) and in a mouse model of EPS, it was demonstrated that macrophages were the immune cells that infiltrated the most and increased the most in the peritoneum during the progression of EPS, while the proportion of infiltrating neutrophils was relatively low (<1%). Flow cytometry also confirmed that cells that produce extracellular traps (CitH3 + ) are also mainly macrophages.
[0032] Therefore, the technical solution of the present application can still prove that intraperitoneal injection of DNase I achieves the effect of treating encapsulating peritoneal sclerosis mainly by inhibiting the formation of extracellular traps of macrophages in mice with encapsulating peritoneal sclerosis.
[0033] Compared with the prior art, this application has the following beneficial effects:
[0034] The present application provides the use of deoxyribonuclease I in the preparation of a drug for treating encapsulating peritoneal sclerosis. The present application finds that deoxyribonuclease I can inhibit METs. Deoxyribonuclease I can effectively reduce the expression of METs in the peritoneal tissue of mice with encapsulating peritoneal sclerosis, and can also significantly improve the abdominal adhesion score, peritoneal inflammatory infiltration, collagen deposition area in the adhesion region, and peritoneal thickness of mice with encapsulating peritoneal sclerosis, thereby improving pathological changes associated with encapsulating peritoneal sclerosis. Therefore, the present application constructs a new prevention and treatment strategy for encapsulating peritoneal sclerosis by targeting METs to treat encapsulating peritoneal sclerosis. BRIEF DESCRIPTION OF THE DRAWINGS
[0035] Figure 1 Single-cell sequencing UMAP map of peritoneal dialysis effluent from EPS patients ( Figure 1 -A to Figure 1 -D is the clinical data and samples taken from peritoneal tissue of healthy control group (n=3), short-term peritoneal dialysis (n=6), long-term peritoneal dialysis (n=4), and peritoneal effluent of EPS patients (n=2); Figure 1 -A is the abdominal CT of the four groups of people, which shows that the peritoneum of EPS patients is thickened (★) and the mesentery is shrunken to form "cauliflower-shaped intestinal loops" (→). Figure 1 -B is a schematic diagram of the single-cell sequencing process for 4 groups of people; Figure 1 -C is the UMAP map of single-cell data from four groups of people; Figure 1 -D is the cell marker gene based on which cells are classified in Figure C, where UPK3B marks mesothelial cells, PDGFRA marks fibroblasts, ACTA2 marks myofibroblasts, EMCN marks endothelial cells, CD68 marks macrophages, CD1C marks monocyte-derived dendritic cells, CD3D marks T lymphocytes, and MS4A1 marks B lymphocytes);
[0036] Figure 2 The results show that the interaction between macrophages and parenchymal cells in EPS increased significantly ( Figure 2 -A is a bar graph showing the intercellular communication intensity values obtained by analyzing the single-cell data of 4 groups of patients through cellchat. The intercellular communication increased significantly with the extension of peritoneal dialysis time, and the highest value was found in EPS; Figure 2-B is the specific changes in cell communication in different groups of patients. The thickness of the arc represents the change in communication intensity. The red arc represents an increase, and the blue arc represents a decrease. The most significant increase in the interaction between parenchymal cells, especially fibroblasts and macrophages in EPS);
[0037] Figure 3 This is the enrichment result of EPS peritoneal macrophages in the extracellular trap formation pathway ( Figure 3 -A: Macrophages were identified into three subtypes: pro-inflammatory macrophages (S100A8+Macro), anti-inflammatory macrophages (C1QA+Macro), and mixed-Macro macrophages in an intermediate state (S100A8+C1QA+Macro, mixed-Macro); Figure 3 -B is the result graph showing a significant increase in the proportion of pro-inflammatory macrophages in the EPS group; Figure 3 -C is a graph showing the differentially expressed genes of three macrophage subtypes in four groups of patients; Figure 3 -D is the KEGG pathway enrichment result diagram of differentially expressed genes in S100A8+ macrophages; Figure 3 -E is the result of GSVA scoring to compare the ETs generation ability of the three macrophage subtypes in the four groups of patients, ***p<0.001);
[0038] Figure 4 The results show that DNase I treatment significantly reduced the expression of METs in the peritoneal cavity ( Figure 4 -A is a schematic diagram of the DNase I treatment of EPS animal experiments: divided into three groups: Control, EPS+Saline, EPS+DNase I; Figure 4 -B is immunofluorescence showing that DNaseI treatment significantly reduced the expression of METs on the peritoneal surface; Figure 4 -C is a statistical graph of MPO-dsDNA concentration in peritoneal lavage fluid measured by ELISA; **p<0.01);
[0039] Figure 5 The results of DNase I treatment to improve the abdominal adhesion and inflammation in EPS mice ( Figure 5 -A: Row a: Small animal ultrasound detection of abdominal adhesion and intestinal peristalsis in living mice; Row b: Naked visual observation of abdominal adhesion; Figure 5 -B is a statistical chart of macroscopic abdominal adhesion scores; Figure 5 -C is the ELISA quantitative detection of IL-6 concentration in peritoneal lavage fluid; *p<0.05, **p<0.01);
[0040] Figure 6 The results of DNase I treatment to improve peritoneal thickening and fibrosis in EPS mice ( Figure 6-A is a histopathological comparison of EPS mice with or without DNase I treatment: row a is a low-power view of the abdominal cross-section showing the severity of intra-abdominal organ adhesions; row b is a MASSON of the adhesion area + Staining images; rows c and d are the thickness of the parietal peritoneum and visceral peritoneum, respectively; rows e and g are the results of immunohistochemistry, showing the differences in the expression of ECM-related proteins in the parietal peritoneum, namely α-SMA, COL1A1 and Fibronectin; Figure 6 -B shows the MASSON in the adhesion area of the three groups of mice + Area statistics chart; Figure 6 -C is the statistical diagram of the subcutaneous thickness of the parietal peritoneum; Figure 6 -D is the statistical diagram of the thickness of the visceral peritoneum, *p<0.05, **p<0.01);
[0041] Figure 7 The figure shows the result that DNase I treatment did not produce cytotoxicity (*p<0.05). DETAILED DESCRIPTION
[0042] In order to better illustrate the purpose, technical solutions and advantages of the present application, the present application will be further described below in conjunction with the accompanying drawings and specific embodiments.
[0043] In the following examples, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used are all commercially available unless otherwise specified, and the components and raw materials used in each parallel experiment are of the same type.
[0044] Example 1
[0045] EPS is diagnosed based on the patient's medical history, clinical symptoms, peritoneal function, and imaging findings.
[0046] like Figure 1 As shown in Figure 2, abdominal CT showed that compared with healthy controls and PD patients without EPS, EPS patients had diffuse peritoneal thickening and "cauliflower-like" intestinal loops ( Figure 1 -A).
[0047] Peritoneal dialysis effluent from 2 patients diagnosed with EPS was collected for single-cell sequencing, and integrated with the single-cell sequencing data of peritoneal dialysis effluent from ordinary peritoneal dialysis patients previously published by the research group (GSE130888) (Si M, Wang Q, Li Y, et al. Inhibition of hyperglycolysis in mesothelial cells prevents peritonealfibrosis. Sci Transl Med. 2019; 11(495):eaav5341.doi:10.1126 / scitranslmed.aav5341), a total of 100,843 cells ( Figure 1 -B).
[0048] Figure 1 -C shows the UMAP map of single-cell data from healthy peritoneal tissue (Healthy Control, HC, n = 3), short-term peritoneal dialysis (SPD, n = 6), long-term peritoneal dialysis (LPD, n = 4), and peritoneal effluent from EPS patients (n = 2) with normal renal function. Figure 1 -D Figure shows the marker genes used for cell classification. This application identified a total of 8 cell types, including mesothelial cells (Mes), fibroblasts (Fib), myofibroblasts (Myofib), endothelial cells (Endo), macrophages (Macro), monocyte-derived dendritic cells (monoDC), T lymphocytes (T cell), and B lymphocytes (B cell).
[0049] The peritoneal cavity is a complex environment composed of multiple cells, and many pathophysiological functions are achieved through cell-cell interactions. In order to explore the causes of changes in parenchymal cells (mesothelial cells, fibroblasts) in the EPS peritoneal environment, the R software package cellchat (Jin S, Guerrero-Juarez CF, Zhang L, et al. Inference and analysis of cell-cell communication using CellChat. Nat Commun. 2021; 12(1): 1088. Published 2021Feb 17. doi: 10.1038 / s41467-021-21246-9) was used to further analyze the interactions of various cells in single-cell sequencing.
[0050] like Figure 2As shown, Figure 2 -A is a bar graph showing the intercellular communication intensity values obtained by analyzing the single cell data of 4 groups of patients through CellChat. Intercellular communication increased significantly with the extension of peritoneal dialysis time, and was the highest in EPS. Figure 2 -B is a circle plot showing the specific changes in cell communication in different groups of patients. The thickness of the arc represents the change in communication intensity. The red arc represents an increase, and the blue arc represents a decrease. In EPS, the increase in the interaction between parenchymal cells, especially fibroblasts and macrophages, is the most significant.
[0051] The analysis showed that the degree of fibrosis gradually increased from the beginning of peritoneal dialysis to the occurrence of EPS. This process was accompanied by the enhancement of cell-to-cell interactions, reaching the highest level in the EPS group. Among them, the increase in the interaction between macrophages and fibroblasts was the most significant. This result suggests that during the formation of EPS, macrophages may affect the transformation of parenchymal cells into fibrosis-related phenotypes by interacting with parenchymal cells (mesothelial cells, fibroblasts).
[0052] The extracted macrophages were clustered again to obtain three macrophage subsets. Combining their strongest specific genes and the definition of macrophage function in previous literature (literature: Yao W, Chen Y, Li Z, et al. Single Cell RNA Sequencing Identifies a Unique Inflammatory Macrophage Subset as a Druggable Target for Alleviating Acute Kidney Injury. Adv Sci (Weinh). 2022; 9(12): e2103675. doi: 10.1002 / advs.202103675), they can be divided into pro-inflammatory macrophages (S100A8+Macro), anti-inflammatory macrophages (C1QA+Macro), and mixed-like macrophages in an intermediate state (S100A8+C1QA+Macro, mixed-Macro) ( Figure 3 -A). The results showed that the proportion of S100A8+ pro-inflammatory macrophages in EPS patients increased significantly, suggesting that macrophages in the EPS environment switch to a pro-inflammatory phenotype ( Figure 3 -B).
[0053] Further differential gene analysis was performed on the three macrophage subtypes from the four groups of patients ( Figure 3-C), and KEGG pathway enrichment analysis was performed on the specific expression genes of S100A8+ macrophages that were significantly increased in the EPS group. We found that they were significantly enriched in pathways such as chemokines, neutrophil extracellular trap (NETs) formation, apoptosis, and inflammation ( Figure 3 -D); GSVA was performed to evaluate the ability of the three macrophage subtypes to form ETs, and it was found that the S100A8+ macrophages in the EPS group had the highest score ( Figure 3 -E), suggesting that the production of extracellular traps (METs) in pro-inflammatory macrophages may be involved in the occurrence and development of EPS.
[0054] Example 2
[0055] This example is a preliminary in vivo animal treatment plan, which can provide a preliminary experimental basis for subsequent in vivo treatment in humans.
[0056] Specific implementation:
[0057] ① Preparation of DNase I solution for animal treatment experiments: Dissolve DNase I dry powder in ddH2O as solvent to prepare 5 mg / mL DNase I solution, shake thoroughly to dissolve and store at -20℃ for later use. In animal experiments, the working dose for the treatment of EPS is 5 mg / kg.
[0058] ②Establishment of EPS mouse model and DNase I treatment: Male 8-week-old C57BL / 6 mice were purchased and used to construct the chemical encapsulating peritoneal sclerosis (EPS) animal model.
[0059] SHS solution (containing 15% ethanol and 0.1% glucose chlorhexidine) was prepared in advance for modeling. In this scheme, the model was divided into a saline group, an EPS group, and an EPS+DNase I treatment group. The model group was intraperitoneally injected with SHS solution (10 mL / kg) every other day for EPS modeling; before the injection of SHS, an equal volume of Saline or DNase I was injected into the peritoneal cavity half an hour in advance, and the mouse's abdominal cavity was gently rubbed to allow the drug to diffuse in the abdominal cavity; while the Saline group was injected with the corresponding volume of saline throughout the whole process, which lasted for 6 weeks. In the late stage of EPS modeling, because adhesions had appeared in the abdominal cavity of some mice, it was necessary to avoid inserting the needle into the dilated intestinal cavity or other organs when injecting drugs intraperitoneally. Before inserting the needle, the abdomen should be gently massaged with fingers to find a suitable insertion site.
[0060] ③ Evaluation of therapeutic effect: After the modeling (6 weeks), the mice were subjected to in vivo abdominal ultrasound examination to evaluate abdominal adhesion and intestinal peristalsis. The mice were then killed for macroscopic adhesion scoring, and the peritoneum was tested by immunofluorescence, immunohistochemistry, ELISA and other methods to evaluate the therapeutic effect of DNase I on EPS. The experimental methods and corresponding results are as follows:
[0061] like Figure 4 As shown in Figure 2, this study used an animal model to explore the feasibility of using DNase I to treat EPS ( Figure 4 -A). The results showed that intraperitoneal administration of DNase I could significantly inhibit the production of METs on the peritoneal surface ( Figure 4 -B), reducing the concentration of MPO-dsDNA in peritoneal lavage fluid ( Figure 4 -C).
[0062] Small animal ultrasound was used to detect abdominal adhesions and intestinal motility in living mice. Figure 5 shown.
[0063] Compared with the mice in the Saline group, DNase I treatment significantly improved the adhesions between the abdominal organs and between the abdominal organs and the parietal peritoneum, and alleviated the symptoms of intestinal obstruction such as slowed intestinal peristalsis and enlarged intestinal cavity ( Figure 5 -A), reduced adhesion score ( Figure 5 -B) and IL-6 levels in peritoneal lavage fluid ( Figure 5 -C), indicating that ablation of METs structure by DNase I reduced the level of peritoneal inflammation in EPS and improved peritoneal adhesions.
[0064] Histopathological examination also compared the differences in abdominal adhesion and fibrosis deposition between different groups of mice. MASSON staining of the mouse abdominal cross-section showed that the degree of adhesion of the abdominal organs in the DNase I treatment group was significantly improved compared with the Saline group ( Figure 6 -A, row a), collagen deposition was significantly reduced in the adhesion area ( Figure 6 - row b in A, Figure 6 -B).
[0065] DNase I treatment also significantly reduced the thickness of the parietal peritoneum and visceral peritoneum in EPS mice ( Figure 6 - row c in A - row d, Figure 6 -C and Figure 6 -D), and the deposition of ECM-related proteins such as α-SMA, COL1A1, and Fibronectin on the peritoneal surface ( Figure 6-e-g in A). The above results suggest that ablating the METs produced in the peritoneal cavity by DNase I treatment can effectively alleviate peritoneal fibrosis and ECM deposition on the peritoneal surface, and ultimately improve the formation of intra-abdominal organ adhesions.
[0066] like Figure 7 As shown, the CCK8 experiment compared the cell activity of peritoneal primary mesothelial cells after exposure to therapeutic doses of DNase I and LiCl. Compared with the control group, DNase I had no significant effect on cell activity, which was better than LiCl (*p<0.05).
[0067] Currently, there are few treatment options for EPS. This application mainly focuses on inhibiting the pathological response of macrophages during EPS—the formation of extracellular traps, which can improve the pathological changes of peritoneal parenchymal cells. Due to the use of DNase I, a drug that specifically inhibits extracellular traps, this regimen has better clinical applicability and lower toxicity than the lithium chloride regimen of Rebecca Herzog et al., and can reduce related treatment side effects.
[0068] Deoxyribonuclease I (DNase I) is a naturally occurring enzyme in living organisms, and is characterized by high safety and low toxicity. At the same time, because the peritoneum is in a state of sterile inflammation during conventional peritoneal dialysis and EPS, the use of this drug to ablate the existing extracellular trapping network structure in the peritoneal cavity will not cause peritoneal infection, providing a better choice for peritoneal immunotherapy, which can prevent or improve EPS peritoneal adhesions while minimizing the risk of infection and hepato-renal toxicity.
[0069] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present application rather than to limit the scope of protection of the present application. Although the present application has been described in detail with reference to the preferred embodiments, ordinary technicians in this field should understand that the technical solution of the present application can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present application.
Claims
1. Application of deoxyribonuclease I in the preparation of inhibitors of macrophage extracellular traps.
2. Application of deoxyribonuclease I as an extracellular trap inhibitor in the preparation of drugs for the treatment of encapsulating peritoneal sclerosis.
3. The use according to claim 2, characterized in that The extracellular trap inhibitor is a macrophage extracellular trap inhibitor.
4. The use according to claim 2, characterized in that The deoxyribonuclease I is prepared into a deoxyribonuclease I solution by using deionized water as solvent.
5. The use according to claim 4, characterized in that The concentration of deoxyribonuclease I in the deoxyribonuclease I solution is 5 mg / mL.
6. The use according to claim 2, characterized in that The deoxyribonuclease I solution is stored at a temperature of -20°C to -30°C for later use, and the deoxyribonuclease I solution is injected intraperitoneally.
7. The use according to claim 2, characterized in that The effective dosage of the deoxyribonuclease I used in animal experiments is 5 mg / kg.
8. The use according to claim 2, characterized in that The encapsulating peritoneal sclerosis includes the encapsulating peritoneal sclerosis caused by peritoneal dialysis or abdominal surgery.
9. The use according to claim 2, characterized in that The deoxyribonuclease I specifically inhibits the macrophage extracellular traps, thereby treating encapsulating peritoneal sclerosis.
10. The use according to claim 2, characterized in that The application includes at least one of the following aspects: 1) DNase I significantly reduced the expression of METs in the peritoneal cavity; 2) Deoxyribonuclease I improves peritoneal adhesion and inflammation in mice with encapsulated peritoneal sclerosis; 3) Deoxyribonuclease I improves peritoneal thickening and fibrosis in mice with encapsulating peritoneal sclerosis.