Application of disulfiram medicine in improving sepsis-induced cardiomyopathy

Abstract

The invention discloses an application of a disulfiram medicine in improving sepsis induced cardiomyopathy. The disulfiram drug is administered in a sepsis mouse model, and the disulfiram drug is proved to have obvious anti-inflammatory effect and myocardial protection effect. The disulfiram medicine can relieve myocardial hypertrophy of mice, relieve the degree of myocardial fibrosis and reduce myocardial interstitial inflammatory exudation, ZO-1 reconstructs continuous linear distribution after administration, and the degree of myocardial edema is obviously relieved. The invention provides a new application of the disulfiram medicine in improving sepsis-induced cardiomyopathy, and provides a new strategy for myocardial protection of sepsis.

Description

TECHNICAL FIELD

[0001] The present application relates to the application of disulfiram in improving sepsis-induced cardiomyopathy, belonging to the technical field of biological medicine. BACKGROUND

[0002] Disulfiram (DSF, molecular formula C 10 H 20 N2S4) is a white to off-white crystalline powder with a slight sulfur smell. It is relatively stable at room temperature, but easily decomposes under high temperature or strong light; it is slightly soluble in water, slightly soluble in ethanol, and easily soluble in organic solvents such as chloroform and diethyl ether. As an artificially synthesized drug, it is usually prepared by reacting tetraethylthiuram disulfide with dimethylamine.

[0003] Disulfiram is an acetaldehyde dehydrogenase (ALDH) inhibitor that mainly targets ALDH2 isomers. By forming a covalent bond with the cysteine residues in the active site of ALDH2, disulfiram can irreversibly inhibit its activity. Ethanol metabolism in the body includes two steps: first, it is converted to acetaldehyde by alcohol dehydrogenase (ADH), and then it is converted to acetic acid under the action of acetaldehyde dehydrogenase (ALDH), especially ALDH2. Disulfiram blocks the conversion of acetaldehyde to acetic acid by inhibiting ALDH2, leading to the accumulation of acetaldehyde and causing facial flushing, headache, nausea, vomiting, palpitations, and other discomforts, which serve as a negative reinforcement mechanism to aid in alcohol cessation and has been approved by the FDA for the treatment of chronic alcohol dependence. Disulfiram can also interact with dopamine beta-hydroxylase (DBH) and P450 enzymes, but its clinical significance is still unclear.

[0004] In recent years, disulfiram has attracted widespread attention due to its anti-tumor activity and anti-inflammatory effects. In cancer treatment, disulfiram induces cancer cell apoptosis by promoting the generation of oxidative stress (ROS), but in inflammation treatment, it can reduce the degree of oxidative stress. This seemingly contradictory effect is actually related to the mechanism of action in different environments.

[0005] Disulfiram exerts an anti-tumor effect by specifically inducing oxidative stress in cancer treatment. Its metabolite, diethyl dithiocarbamate (DTC), forms a DTC-Cu complex with copper ions, which inhibits superoxide dismutase (SOD) activity and interferes with the mitochondrial electron transport chain, leading to a burst of reactive oxygen species (ROS) accumulation. In addition, DTC-Cu can also inhibit 26S proteasome function, leading to the accumulation of misfolded proteins and triggering endoplasmic reticulum stress (ERS), further amplifying the ROS signal. High concentrations of ROS break through the antioxidant defense threshold of cancer cells, activate pro-apoptotic pathways such as p53 / JNK / MAPK, and inhibit survival signals such as NF-κB and Nrf2, ultimately selectively inducing tumor cell apoptosis.

[0006] In inflammatory diseases, disulfiram reduces the level of oxidative stress through multi-target regulation. Low concentration of DSF directly inhibits the nuclear translocation of NF-κB and the activation of NLRP3 inflammasome, blocks the release of pro-inflammatory factors (such as TNF-α, IL-1β); at the same time, it activates the Keap1 / Nrf2 pathway, up-regulates the expression of antioxidant proteins such as glutathione synthetase (GCLC / GCLM), heme oxygenase-1 (HO-1), and inhibits the activity of NADPH oxidase (NOX), reducing the level of intracellular ROS. This synergistic effect reshapes the redox balance and reduces the free radical damage driven by immune cells. In addition, DSF can also reduce the inflammatory response by inhibiting angiogenesis (such as reducing the production of TNF-α and VEGF).

[0007] Sepsis is a systemic inflammatory response syndrome triggered by infection, with a mortality rate of 30%-70%. Its pathological process begins with an uncontrolled inflammatory storm and eventually progresses to multiple organ dysfunction. Sepsis-induced cardiomyopathy (SIC) is a critical complication of sepsis, characterized by myocardial contractile dysfunction and progressive myocardial edema-fibrosis remodeling, significantly worsening patient prognosis. The STING pathway triggers the release of type I interferon (IFN-1) and inflammatory factors by recognizing bacterial / host-derived DNA in sepsis, thereby activating endothelial cells. Activated endothelial cells rapidly release angiopoietin-2 (Ang-2) and von Willebrand factor (vWF) from Weibel-Palade bodies, leading to increased vascular permeability, capillary leakage, and microthrombosis, exacerbating myocardial edema and causing severe cardiac dysfunction.

[0008] Existing studies have confirmed that disulfiram reduces systemic inflammation in sepsis by inhibiting the NF-κB pathway, but its targeted regulation mechanism and effects on myocardial edema have not been clearly defined. SUMMARY

[0009] The purpose of the present application is to provide the application of disulfiram drug in improving sepsis-induced cardiomyopathy.

[0010] To achieve the above-mentioned purpose, the present application provides the application of disulfiram drug in preparing a drug for treating or alleviating sepsis-induced cardiomyopathy or related conditions.

[0011] Preferably, the sepsis-induced cardiomyopathy-related conditions include myocardial edema caused by sepsis.

[0012] Preferably, the drug comprises an effective ingredient and a pharmaceutically acceptable carrier or excipient, and the effective ingredient is disulfiram.

[0013] Preferably, the dosage form of the drug is injection, tablet, powder, suspension or capsule.

[0014] Compared with the prior art, the present application has the following beneficial effects:

[0015] The present application provides a new use of disulfiram in improving sepsis-induced cardiomyopathy, focuses on the molecular mechanism of disulfiram targeting to improve sepsis cardiomyopathy myocardial edema, and shows that disulfiram has obvious anti-inflammatory effect and myocardial protection effect by administering disulfiram to a sepsis mouse model; disulfiram can relieve myocardial hypertrophy, reduce the degree of myocardial fibrosis, and reduce inflammatory exudation in myocardial interstitium, and after administration, ZO-1 is reestablished in a continuous linear distribution, and the degree of myocardial edema is significantly reduced; that is, disulfiram intervenes in the key link of edema formation in multiple dimensions, and the present application provides a new strategy for sepsis myocardial protection. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 The spleen of the model group and the control group is taken for a photo;

[0017] Figure 2 The lung Masson staining result is shown in the following table:

[0018] Figure 3 The liver Masson staining result is shown in the following table:

[0019] Figure 4 The kidney Masson staining result is shown in the following table:

[0020] Figure 5 The heart of the model group and the administration group is taken for a photo;

[0021] Figure 6 The myocardial WGA staining result is shown in the following table:

[0022] Figure 7 The myocardial Masson staining result is shown in the following table:

[0023] Figure 8 The myocardial HE staining result is shown in the following table:

[0024] Figure 9 The ZO-1 protein immunohistochemistry is shown in the following table: DETAILED DESCRIPTION

[0025] In order to make the present application more obvious and easy to understand, the preferred embodiments are described in detail below with reference to the accompanying drawings.

[0026] The experimental methods not specified in the following examples are generally carried out according to conventional conditions or according to the conditions recommended by the manufacturer. The materials, reagents, etc. used in the following examples are commercially available reagents and materials unless otherwise specified.

[0027] EXAMPLE

[0028] This study investigated the protective effect of disulfiram against septic cardiomyopathy using a mouse cecal ligation (CLP) model of sepsis. Eighteen 8-10 week old male C57BL / 6 mice were randomly divided into three groups based on similar weight and age: a Control group, a CLP group, and a CLP+DSF group. The CLP and CLP+DSF groups underwent CLP to establish a sepsis model, while the Control group received no treatment. The CLP+DSF group received 50 mg / kg of disulfiram solution daily for four consecutive days prior to surgery, while the CLP and Control groups received an equal volume of saline.

[0029] After the model was established, all mice were fed for 2 weeks, and then the heart, liver, spleen, lungs and kidneys were removed for tissue fixation and pathological sectioning.

[0030] The results showed that the spleen size of mice in the CLP+DSF group was smaller than that in the CLP group. Figure 1 As shown, but slightly larger than the Control group. Pathological examination of sections revealed that, compared with the CLP group, the CLP+DSF group mice showed significantly reduced inflammatory responses in the lungs, liver, and kidneys, as shown in the figure. Figures 2 to 4 The Masson staining results for each tissue are shown. This indicates that the cecal ligation and perforation model was successfully established, the CLP group showed a significant inflammatory immune response, and disulfiram had a significant anti-inflammatory effect.

[0031] In a cecal ligation and puncture (CLP) sepsis model, the CLP+DSF group exhibited a clear cardioprotective effect. From a macroscopic morphological perspective, the CLP+DSF group showed a significant reduction in heart volume and ventricular wall thickness, suggesting a alleviation of myocardial hypertrophy. Figure 5 As shown in the figure. WGA staining results showed that the cross-sectional area of ​​cardiomyocytes was increased in the CLP group, while the CLP+DSF group was close to the normal level. Figure 6 As shown; Masson staining revealed an increased area of ​​blue collagen deposition in the interstitial matrix of the model group, while the degree of fibrosis was reduced in the CLP+DSF group, as shown. Figure 7 As shown; HE staining results also indicated that the CLP group had a large amount of inflammatory exudate in the myocardial interstitium and perivascular "cuff-like" edema, while the CLP+DSF group had reduced exudate, as shown. Figure 8 As shown in the immunohistochemical results, the CLP group exhibited fragmented and dissociated expression of the microvascular tight junction protein (ZO-1), accompanied by inflammatory infiltration and interstitial edema. In contrast, the CLP+DSF group showed reconstructed continuous linear distribution of ZO-1, and reduced myocardial edema. Figure 9 As shown.

[0032] To further verify the transcriptional regulation of disulfiram (DSF) on the "STING-Ang-2 / vWF-ZO-1" axis, we quantitatively analyzed the expression levels of key target genes in the myocardial tissue of CLP mice by real-time fluorescence quantitative PCR (qPCR). The myocardial tissue of the Control, CLP and CLP+DSF groups at 14 days after the operation was taken, total RNA was extracted by Trizol method, and cDNA was synthesized by reverse transcription. The verified qPCR primers (Ang-2, vWF, internal reference GAPDH) were selected for amplification, and the reaction system and procedure followed the standard process of SYBR Green: pre-denaturation at 95°C for 30s; 95°C for 5s, 60°C for 30s, 40 cycles; melting curve analysis to confirm the specificity of the product. Taking GAPDH as the internal reference, the relative expression was calculated by the 2^–ΔΔCt method.

[0033] The qPCR detection confirmed that disulfiram inhibited the transcriptional expression of Ang-2 / vWF. The mRNA expression levels of Ang-2 and vWF in the myocardial tissue of the CLP group were significantly increased, indicating that sepsis induced endothelial cell activation and Weibel-Palade body release. The expression levels of Ang2 and vWF in the CLP+DSF group were lower than those in the CLP group, but were still higher than those in the control group. The results confirmed from the transcriptional level that disulfiram significantly down-regulated the mRNA levels of Ang-2 and vWF by inhibiting the STING pathway, thereby blocking the capillary leakage mediated by the "Ang-2 / vWF-ZO-1" axis and reducing sepsis-induced myocardial edema, which was consistent with the phenotype of ZO-1 connection repair and WGA staining edema reduction in immunohistochemistry.

[0034] The present application proposes a new use of disulfiram in the treatment of sepsis-induced cardiomyopathy, which intervenes in sepsis-induced cardiomyopathy (SIC) myocardial edema through the "STING-Ang-2 / vWF-ZO-1" axis multi-target. Specifically, disulfiram can inhibit the STING pathway, block endothelial cell activation and Weibel-Palade body release of Ang-2 / vWF; stabilize the ZO-1 / VE-cadherin tight junction, reduce capillary leakage; reduce the systemic inflammatory storm, and synergistically reduce the volume of protein-rich interstitial fluid in the myocardium, thereby relieving edema-fibrosis remodeling and improving cardiac dysfunction.

[0035] The above is only a preferred embodiment of the present application, and is not a limitation on the form and substance of the present application. It should be noted that those skilled in the art can make some improvements and supplements without departing from the present application, and these improvements and supplements should also be considered as the protection scope of the present application.

Claims

1. Use of disulfiram in the manufacture of a medicament for treating or ameliorating sepsis-induced cardiomyopathy or a related condition.

2. Use according to claim 1, wherein The sepsis-induced cardiomyopathy-related condition includes myocardial edema caused by sepsis.

3. The use according to claim 1, wherein The medicament comprises an effective ingredient of disulfiram and a pharmaceutically acceptable carrier or excipient.

4. Use according to any one of claims 1 to 3, characterized in that, The dosage form of the medicament is injection, tablet, powder, suspension or capsule.

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