Mononuclear cell for treating sepsis cardiomyopathy and application thereof

Abstract

The invention provides a mononuclear cell for treating sepsis cardiomyopathy, and also relates to application of the mononuclear cell in preparation of drugs for improving sepsis and sepsis cardiomyopathy. The mononuclear cell provided by the invention can have a better targeted enrichment characteristic, so that the effective rate of treatment is improved, and meanwhile, toxic and side effects caused by non-specific distribution of drugs are reduced. Besides, the mononuclear cells are derived from the body of a healthy body, so that the mononuclear cells are more stable in metabolism, higher in absorption efficiency and more excellent in enrichment effect when acting on the body, and the technical problems that in-vivo half-life period of drugs is short and catabolism is easy in the prior art are solved.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a monocyte for treating septic cardiomyopathy, and also relates to the use of monocytes in preparing drugs for improving sepsis and septic cardiomyopathy. Background Art

[0002] Sepsis is a life-threatening disease caused by a dysregulated host immune response to infection, often manifesting as rapidly progressive organ dysfunction and high mortality. The incidence of sepsis is particularly high in intensive care units (ICUs). Its pathological mechanisms are extremely complex, involving multiple components, including the immune system, inflammatory responses, microcirculatory dysfunction, and cellular metabolic imbalance. During the progression of sepsis, cytokines and endotoxins (such as lipopolysaccharide) released by pathogens activate the host immune system, leading to overactivation of immune cells and the release of inflammatory mediators. While these responses may help control infection in the short term, excessive inflammation can cause endothelial damage, leading to microcirculatory impairment, hemodynamic instability, and tissue hypoxia, exacerbating organ failure and causing an immunosuppressive state, increasing the risk of secondary infection. Sepsis is often accompanied by septic cardiomyopathy, which occurs in approximately 60% of patients with sepsis or septic shock and carries a mortality rate of 70% to 90%. Clinical manifestations of septic cardiomyopathy typically include decreased ejection fraction, left ventricular dilation, and early, reversible myocardial dysfunction. Septic cardiomyopathy is not only one of the main causes of multiple organ failure but also a significant factor in poor prognosis and mortality in patients with sepsis. Therefore, research on the treatment of septic cardiomyopathy is particularly important, especially finding treatments with minimal side effects and strong targeting.

[0003] Currently, the treatment of sepsis primarily involves early anti-infective therapy, hemodynamic support, oxygen therapy, renal replacement therapy, and immune response regulation. Clinical treatment for septic cardiomyopathy, however, still focuses on suppressing cardiac inflammation, reducing cardiac workload, and enhancing myocardial contractility, but targeted therapeutic options are lacking. Therefore, in-depth research into the pathogenesis of sepsis, particularly septic cardiomyopathy, and the exploration of new therapeutic targets are crucial for improving patient survival and therapeutic outcomes. Summary of the Invention

[0004] In response to the above technical problems, the present invention provides a monocyte for treating septic cardiomyopathy, and also provides the use of monocytes in preparing drugs for improving sepsis and septic cardiomyopathy.

[0005] In order to achieve the above-mentioned object of the invention, the technical solution of the present invention includes: The present invention first provides a monocyte for treating septic cardiomyopathy. The monocyte is a monocyte treated with sodium lactate in vitro.

[0006] The present invention has expanded new approaches to shaping monocyte function. Specifically, the use of sodium lactate to acclimate monocytes has the following unique features: (1) Metabolic reprogramming: Sodium lactate alters the phenotype and function of monocytes by affecting cellular metabolic pathways (e.g., lactate metabolism and mitochondrial function). This strategy of shaping cellular function through metabolic regulation is still a gap in the existing technology. (2) Histone lactylation: Sodium lactate induces histone lactylation in monocytes, thereby regulating monocyte gene expression and endowing the cells with new functional properties.

[0007] Treating sodium lactate-acclimated monocytes in vitro and then reinfusing them is a unique cell therapy model. Its innovations are reflected in the following aspects: (1) Enhanced targeting: Through in vitro acclimation, monocytes are given specific functions (promoting repair), improving their specificity and effectiveness in treating septic cardiomyopathy in vivo. (2) Avoiding side effects of direct administration: Direct use of sodium lactate in the body may cause systemic metabolic disorders, but using cells as "carriers" to indirectly exert the therapeutic effects of monocytes can reduce the risk of side effects. The in vitro and in vivo combined model of the present invention enhances the therapeutic effect of sodium lactate while avoiding the side effects that may be caused by direct drug application.

[0008] Furthermore, the in vitro treatment process includes the following steps, S1: Isolate mononuclear cells from tissues or body fluids by density gradient centrifugation; S2: The monocytes isolated in step S1 were suspended in RPMI-1640 medium containing 5-20 mM sodium lactate and cultured in an incubator at 36-38°C and 5%-10% CO2 for 24 hours.

[0009] Sodium lactate, as the sodium salt of lactic acid, has a milder acidity than lactic acid. It can improve sepsis-related metabolic imbalances through a new post-translational modification called lactation, and plays an important role in cell therapy. Sodium lactate promotes the effective function of cells in the cardiac microenvironment through lactation modification, and enhances the effect of cell therapy through mechanisms such as improving microcirculation, reducing myocardial cell apoptosis, and regulating immune responses. It shows unique clinical potential, especially in the treatment of sepsis-induced heart failure. Therefore, the application of sodium lactate in sepsis cell therapy not only helps to improve cardiac function, but also promotes the improvement of treatment effects by regulating metabolism and immune responses, becoming a new direction for the future treatment of sepsis.

[0010] Furthermore, in step S2, the RPMI-1640 culture medium contains 5-10% fetal bovine serum and 1%-2% penicillin-streptomycin.

[0011] Furthermore, the monocytes are derived from one or more of peripheral blood, spleen and bone marrow.

[0012] Based on this, the present invention also provides the use of the monocytes in the preparation of drugs for improving sepsis and septic cardiomyopathy.

[0013] Application expansion of the present invention for septic cardiomyopathy This invention, based on metabolically regulated monocyte therapy, offers a novel therapeutic strategy for septic cardiomyopathy. While current treatments for septic cardiomyopathy primarily focus on supportive therapies or targeted therapies targeting inflammatory factors, this invention focuses on the metabolic abnormalities underlying septic cardiomyopathy: 1) Correcting monocyte dysfunction through metabolic remodeling offers a novel perspective for the treatment of septic cardiomyopathy. 2) Potential cardioprotective effects: After acclimating monocytes with sodium lactate, the treatment may exert its effects by inhibiting cardiac inflammation or promoting myocardial repair.

[0014] This invention improves the safety and durability of treatment, opening up a new avenue for cell therapy. In vitro acclimation followed by reinfusion of monocytes offers the following advantages: 1) Longer-lasting efficacy: Treated monocytes can continue to exert their regulatory effects without relying on the continuous infusion of exogenous drugs. 2) Optimized safety: Systemic drug exposure is reduced, while potentially toxic metabolites within the cells are reduced through in vitro conditioning.

[0015] This invention enables customized and precise cell therapy, tailored to the specific conditions of each patient. By adjusting the sodium lactate concentration and acclimation time based on the patient's specific condition, personalized treatment is achieved. This flexibility is difficult to achieve with existing fixed-dose drug therapies.

[0016] Furthermore, monocytes are used in the preparation of drugs for improving cardiac function in septic cardiomyopathy, wherein cardiac function includes left ventricular ejection fraction and left ventricular fractional shortening.

[0017] Furthermore, mononuclear cells are isolated from one or more of peripheral blood, spleen and bone marrow.

[0018] Compared with the prior art, the beneficial effects of the present invention are embodied in:

[0019] (1) After the occurrence of sepsis, especially when complicated by septic cardiomyopathy, a large number of mononuclear macrophages are recruited in the heart. After in vitro sodium lactate stimulation, the mononuclear cells are also recruited to the heart along with the blood circulation to play a therapeutic role. Compared with the traditional treatment strategy of inhibiting cardiac inflammatory response, reducing cardiac load, and enhancing myocardial contractile function, this cell therapy has better targeted enrichment characteristics, thereby improving the treatment efficiency and reducing the toxic side effects caused by the non-specific distribution of drugs. In addition, since the monocytes of the present invention are derived from the body of a healthy organism, when they act on the body, their metabolism is more stable, the absorption efficiency is higher, and the enrichment effect is better, which solves the technical problems of low half-life and easy decomposition and metabolism of drugs in the prior art.

[0020] (2) The present invention discloses the application of monocytes prepared by sodium lactate for the treatment of sepsis. By constructing a C57 mouse septic cardiomyopathy model, the feasibility of using sodium lactate to prepare monocytes for the treatment of sepsis is explored. The experimental results confirmed that monocyte treatment after sodium lactate stimulation can significantly improve the hypothermia and weight loss caused by sepsis, and significantly improve the cardiac function during septic cardiomyopathy. In addition, the immunofluorescence results showed that monocyte treatment after sodium lactate stimulation can significantly reduce the apoptosis and oxidative stress of myocardial cells during septic cardiomyopathy. These results show that monocyte treatment after sodium lactate stimulation can indeed improve the systemic infection symptoms caused by sepsis, and significantly improve the damage to the heart caused by septic cardiomyopathy, and improve the cardiac function of the heart with septic cardiomyopathy. The present invention proposes and proves for the first time the application of sodium lactate-stimulated monocytes in sepsis, providing a new idea for the clinical treatment of sepsis and septic cardiomyopathy.

[0021] (3) The present invention has expanded new approaches to shaping monocyte function. Specifically, the use of sodium lactate to domesticate monocytes has the following unique features: 1) Metabolic reprogramming: Sodium lactate changes the phenotype and function of monocytes by affecting cellular metabolic pathways (such as lactate metabolism and mitochondrial function). This strategy of shaping cell function through metabolic regulation is still a blank in the existing technology. 2) Histone lactylation: Sodium lactate induces histone lactylation in monocytes, thereby regulating monocyte gene expression and giving cells new functional properties.

[0022] (4) The in vitro and in vivo combined model of the present invention enhances the therapeutic effect of sodium lactate while avoiding the side effects that may be caused by direct drug application. Specifically, the in vitro treatment of monocytes acclimated with sodium lactate and then reinfusion is a unique cell therapy model. The innovation is reflected in: 1) Enhanced targeting: Through in vitro acclimation, monocytes are given specific functions (promoting repair), thereby improving their specificity and effectiveness in treating septic cardiomyopathy in vivo. 2) Avoiding the side effects of direct administration: Sodium lactate directly used in the body may cause systemic metabolic disorders, while using cells as "carriers" to indirectly exert the therapeutic effects of monocytes can reduce the risk of side effects.

[0023] (5) Currently, treatments for septic cardiomyopathy mostly focus on supportive therapy or targeted therapy against inflammatory factors. However, this invention focuses on the metabolic abnormalities of septic cardiomyopathy: 1) Correcting monocyte dysfunction through metabolic remodeling provides a new perspective for the treatment of septic cardiomyopathy. 2) Potential cardioprotective effects: After sodium lactate acclimates monocytes, it may exert its effects by inhibiting cardiac inflammation or promoting myocardial repair. Therefore, this invention provides a new treatment strategy for septic cardiomyopathy based on metabolically regulated monocyte therapy.

[0024] (6) This invention improves the safety and durability of treatment, opening up a new avenue for cell therapy. In vitro acclimation followed by reinfusion of monocytes has the following advantages: 1) Longer-lasting therapeutic effects: The treated monocytes can continue to exert regulatory effects without relying on the continuous injection of exogenous drugs. 2) Optimized safety: Reduced systemic drug exposure and the reduction of potentially toxic metabolites within the cells through in vitro treatment.

[0025] (7) The present invention adjusts the sodium lactate concentration and acclimation time according to the patient's specific condition, thereby achieving personalized treatment. This flexibility is difficult to achieve with existing fixed-dose drug therapies. As can be seen, the present invention achieves customized and precise cell therapy, adapting to the specific conditions of different patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a comparison chart of the results of Western blotting experiments after monocytes were stimulated with sodium lactate.

[0027] Figure 2 This is a control chart for PKH26 immunofluorescence staining.

[0028] Figure 3 This is a graph showing the results of body temperature in animal experiments.

[0029] Figure 4 This is a graph showing the body weight results in animal experiments.

[0030] Figure 5This is the cardiac ultrasound result after sodium lactate-stimulated monocyte treatment in animal experiments.

[0031] Figure 6 This is a cardiac immunofluorescence image (TUNEL) after sodium lactate-stimulated monocytes were treated in animal experiments.

[0032] Figure 7 This is a cardiac immunofluorescence image (DHE) after sodium lactate-stimulated monocytes were treated in animal experiments. DETAILED DESCRIPTION

[0033] The technical solution of the present invention is further described in detail below with reference to specific embodiments.

[0034] The present invention provides a mononuclear cell for treating septic cardiomyopathy. The mononuclear cell is a mononuclear cell treated with sodium lactate in vitro.

[0035] Specifically, the in vitro treatment process includes the following steps: S1: Isolate mononuclear cells from tissues or body fluids by density gradient centrifugation; S2: The monocytes isolated in step S1 were suspended in RPMI-1640 medium containing 5-20 mM sodium lactate and cultured in an incubator at 36-38°C and 5%-10% CO2 for 24 hours.

[0036] RPMI-1640 culture medium contains 5-10% fetal bovine serum and 1-2% penicillin-streptomycin. Mononuclear cells are isolated from one or more of peripheral blood, spleen, and bone marrow.

[0037] Also provided is the use of the aforementioned monocytes in the preparation of a drug for ameliorating sepsis and septic cardiomyopathy. Cardiac function includes left ventricular ejection fraction and left ventricular fractional shortening. The monocytes are derived from one or more of peripheral blood, spleen, and bone marrow. Example 1 This example aims to study the potential therapeutic effect of stimulating monocytes with sodium lactate in vitro and then infusing them into mice with septic cardiomyopathy. The specific steps include: S1: Mononuclear cells were isolated from peripheral blood of wild-type mice by density gradient centrifugation; S2: The monocytes isolated in step S1 were resuspended in RPMI-1640 medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin) containing 20 mM sodium lactate and cultured in an incubator at 37°C and 5% carbon dioxide for 24 hours to allow the cells to be pretreated with sodium lactate.

[0038] S3: Then, the lactate-stimulated monocytes were injected into the septic cardiomyopathy mice via tail vein injection.

[0039] Application Example 1 Stimulatory effect of sodium lactate on histone lactylation in monocytes.

[0040] Sodium lactate was added to monocytes, and the cells were divided into three groups according to different sodium lactate concentrations. 5mM sodium lactate, 20mM sodium lactate, and normal saline were added as a control. The cells were cultured for 24 hours, and the cell histones were extracted to detect the expression of histone lactylation in monocytes.

[0041] Depend on Figure 1 As shown in the immunoblotting results, sodium lactate has a significant stimulating effect on monocyte histone lactylation. Application Example 2 Animal experiments verified the therapeutic effect of sodium lactate-stimulated monocyte therapy on septic cardiomyopathy.

[0042] 1. A septic cardiomyopathy model was established, and mice were divided into three groups. Mononuclear cells stimulated with sodium lactate, mononuclear cells not stimulated with sodium lactate, and normal saline were injected into the tail vein, respectively.

[0043] 2. Add PKH26 (a cell membrane dye) to monocytes stimulated with sodium lactate and monocytes not stimulated with sodium lactate, detect PKH26 immunofluorescence staining in frozen heart sections, and observe the infiltration of mononuclear macrophages in cardiac tissue after sodium lactate stimulation.

[0044] Depend on Figure 2 As shown in the immunofluorescence results in the middle figure, sodium lactate-stimulated monocyte transfusion can enter the heart and play a role in septic cardiomyopathy.

[0045] 18 hours after the onset of sepsis, the body temperature and weight of the mice were measured, cardiac function was assessed by cardiac ultrasound, and myocardial cell apoptosis and oxidative stress levels were assessed by frozen sections. Figure 3-7 .

[0046] Depend on Figure 3-7 It can be seen that the treatment of mononuclear cells stimulated by sodium lactate can significantly improve the hypothermia and low body weight caused by sepsis, and significantly improve the damage to the heart caused by sepsis cardiomyopathy, and improve the cardiac function of the heart with sepsis cardiomyopathy. The body temperature, body weight, cardiac ejection fraction (EF%), and left ventricular short axis shortening (FS%) of mice treated with mononuclear cells stimulated by sodium lactate were significantly higher than those of the control group ( Figure 3-5 ), while myocardial cell apoptosis and oxidative stress were significantly lower than those in the control group ( Figure 6-7 ).

[0047] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Any simple modification, equivalent change, and modification based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A mononuclear cell for treating septic cardiomyopathy, characterized in that: The monocytes are monocytes treated in vitro with sodium lactate.

2. The mononuclear cells for treating septic cardiomyopathy according to claim 1, characterized in that: The in vitro treatment process comprises the following steps: S1: Isolate mononuclear cells from tissues or body fluids by density gradient centrifugation; S2: The monocytes isolated in step S1 were suspended in RPMI-1640 medium containing 5-20 mM sodium lactate and cultured in an incubator at 36-38°C and 5%-10% CO2 for 24 hours.

3. The mononuclear cells for treating septic cardiomyopathy according to claim 2, characterized in that: In step S2, the RPMI-1640 culture medium contains 5-10% fetal bovine serum and 1%-2% penicillin-streptomycin.

4. The mononuclear cells for treating septic cardiomyopathy according to claim 2, characterized in that: In step S1, mononuclear cells are isolated from one or more of peripheral blood, spleen and bone marrow.

5. Use of the monocytes according to any one of claims 1 to 4 in the preparation of a drug for improving sepsis and septic cardiomyopathy.

6. The use according to claim 5, characterized in that: The monocytes are derived from one or more of peripheral blood, spleen and bone marrow.

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