Application of 2-ME in preparation of medicine for preventing or treating sepsis cardiomyopathy

Abstract

The invention discloses application of 2-ME in preparation of a medicine for preventing or treating sepsis cardiomyopathy, low-dose 2-ME can recover the activity of myocardial cells damaged by sepsis, reduce the apoptosis rate of the myocardial cells and enhance the contractility of the myocardial cells, so that the heart function is improved; the 2-ME can significantly inhibit cell death, reduce accumulation of disulphide in cells and prevent occurrence of disulphide death, and has potential application prospects in myocardial protection and prevention and treatment of sepsis myocardial injury.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular, relates to the application of 2-ME in preparing a medicine for preventing or treating septic cardiomyopathy. Background Art

[0002] Sepsis is a complex clinical syndrome caused by a dysregulated response of the body to infection, involving abnormal activation of the immune system and inflammatory responses. During the progression of sepsis, patients may suffer end-organ damage, especially to the heart. As one of the important organs of the human body, impaired function of the heart can seriously affect the entire circulatory system and lead to serious health problems. In the cardiovascular manifestations of sepsis, patients usually experience a dynamic process of change. Initially, patients may show a hypodynamic state, which refers to a decrease in the heart's ability to pump blood, resulting in slowed blood circulation. Subsequently, patients may enter a hyperdynamic state, when the heart's output increases and systemic vascular resistance decreases, which may lead to uneven blood flow distribution and insufficient tissue perfusion. During this process, if there is a reversible decrease in cardiac function, that is, the contractility of both the left and right ventricles is weakened, this condition is called septic cardiomyopathy (SICM). SICM is one of the main causes of mortality and morbidity during septic shock and a common complication in patients with sepsis. It is closely associated with high mortality and has a significant impact on the prognosis of patients.

[0003] SICM is characterized by the lack of significant myocardial cell apoptosis in pathology, tissue oxygenation levels usually remain normal, and the organ dysfunction caused by it is reversible after appropriate treatment. This means that if SICM can be identified and treated in time, the patient's cardiac function may be restored to the state before sepsis. However, in clinical practice, the treatment and intervention measures received by patients may affect their physiological parameters, which makes the study of SICM relatively difficult. For example, the use of vasoactive drugs, fluid resuscitation, and mechanical ventilation may change the patient's blood pressure, heart rate, and cardiac output, thereby affecting the diagnosis and evaluation of SICM. In addition, the pathophysiological mechanism of SICM is complex and involves multiple interactions at the cellular and molecular levels, which further increases the complexity of the study.

[0004] Currently, there are no specific treatment guidelines or strategies for SICM. Existing treatments focus on supportive care and infection control, including the use of antibiotics, fluid management, vasoactive drugs, and mechanical ventilation when necessary. However, these treatments are mainly aimed at the general management of sepsis rather than specifically for the treatment of SICM. Therefore, a better understanding of the underlying mechanisms of SICM is crucial for developing targeted treatment strategies and improving patient outcomes. In order to improve the understanding and treatment of SICM, future research needs to focus on the following aspects: First, the pathophysiological mechanisms of SICM need to be further elucidated, including inflammatory mediators, cell signaling pathways, and metabolic changes in cardiomyocytes. Second, diagnostic tools and biomarkers for SICM need to be developed and validated to more accurately identify and evaluate SICM. In addition, clinical trials are needed to evaluate the efficacy of different treatment strategies for SICM, including drug therapy, mechanical circulatory support, and cardiac rehabilitation. Through these studies, if more precise and effective SICM treatment guidelines are developed, the mortality rate of patients with sepsis and their long-term prognosis can be reduced, which is of great significance for the prevention and treatment of SICM.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The technical problem to be solved by the present invention is to overcome the deficiencies of the prior art, provide the use of 2-ME in the preparation of drugs for preventing or treating septic cardiomyopathy, lay a good foundation for the development of new protective drugs for septic cardiomyopathy in terms of mechanism, provide new myocardial protective drugs, reduce the mortality rate of septic patients and improve their long-term prognosis.

[0007] In order to solve the above technical problems, the basic concept of the technical solution adopted by the present invention is:

[0008] 2-Mercaptoethanol (2ME) is a cross-linking agent for disulfide bonds, which can help peptides and proteins form the correct spatial conformation. It is a thiol-containing antioxidant widely used in cell biology research. It can react with cystine to generate cysteine, and then generate glutathione to scavenge free radicals. 2-ME is a compound with a variety of potential health benefits, and it has shown positive effects on health in preclinical models. The most striking of these is its role in tumor suppression. Studies have shown that 2-ME can inhibit the growth and spread of tumor cells by regulating the level of oxidative stress in cells and affecting cell signaling pathways. In addition, 2-ME can also activate the cell's DNA damage response mechanism, help repair damaged DNA, and prevent carcinogenesis caused by DNA damage. In addition to its potential in tumor suppression, 2-ME is also believed to prolong life. In some animal models, 2-ME treatment is associated with an extended average lifespan and delayed aging. This may be due to its powerful antioxidant properties, which help reduce free radical damage to cells, which is a key factor in cell aging and functional decline. In terms of immune function, 2-ME also shows a positive effect. It is able to modulate the activity of immune cells and enhance the body's defense against pathogens. By maintaining a balanced immune system, 2-ME helps improve the body's resistance to infection and disease. Overall, these potential benefits of 2-ME emphasize the importance of further studying its use in preventive medicine and therapeutic applications. With further research, 2-ME may become a powerful tool for the future treatment of a variety of diseases, including cancer and degenerative diseases. However, it is important to note that although laboratory studies have shown positive results, the clinical use of 2-ME requires more research to determine its safety and effectiveness.

[0009] Disulfidptosis is a newly discovered cell death mode that is closely related to 2-ME. Under specific conditions, such as glucose starvation, high expression of SLC7A11 (xCT) leads to the depletion of intracellular NADPH, resulting in abnormal accumulation of disulfides such as cysteine, inducing disulfide stress and ultimately leading to cell death. This cell death mode is different from known cell death types such as apoptosis, necrosis and ferroptosis, and it involves abnormal disulfide bond formation of actin cytoskeleton proteins and the collapse of the F-actin network. 2-ME, as a disulfide bond reducing agent, can effectively inhibit disulfide death. In the experiment, the researchers found that the addition of 2-ME could significantly inhibit H2O2-induced cell death, indicating that 2-ME can reduce the accumulation of intracellular disulfides and prevent the occurrence of disulfide death. In addition, 2-ME and other disulfide bond reducing agents such as TCEP and DTT can completely inhibit the cell death of SLC7A11 high-expressing cells under glucose starvation conditions. These findings suggest that 2-ME and similar disulfide reducing agents may have potential applications in the prevention and treatment of diseases associated with disulfide death. By regulating intracellular disulfide levels, these reducing agents may help protect cells from disulfide death, especially in pathological conditions where glucose supply is limited or NADPH is depleted.

[0010] At present, there is no research on the protective effect of 2-ME on the cell model of myocardial injury in sepsis and its application in the treatment of septic cardiomyopathy. The present invention verifies the protective effect of low-dose 2-ME (20umol / L) on myocardial cell injury in sepsis and the prevention and treatment of septic cardiomyopathy, which can lay a good foundation for the development of new protective drugs for septic cardiomyopathy in terms of mechanism, thereby providing a new myocardial protective drug.

[0011] After adopting the above technical scheme, the present invention has the following beneficial effects compared with the prior art.

[0012] Low-dose 2-ME can restore the activity of myocardial cells damaged by sepsis, reduce the apoptosis rate of myocardial cells, and enhance the contractile ability of myocardial cells, thereby improving cardiac function. We initially found that 2-ME exerts a cardioprotective effect by inhibiting disulfide death. We evaluated the effect of 2-ME through in vitro cell experiments. The experimental results showed that 2-ME can significantly inhibit cell death, reduce the accumulation of intracellular disulfides, and prevent the occurrence of disulfide death. 2-ME has potential application prospects in protecting the myocardium, preventing and treating myocardial damage caused by sepsis.

[0013] The specific implementation modes of the present invention are further described in detail below in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The accompanying drawings are part of this application and are used to provide a further understanding of the present invention. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention, but do not constitute an improper limitation of the present invention. Obviously, the drawings described below are only some embodiments. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. In the drawings:

[0015] Figure 1 This is the construction of the LPS-induced septic cardiomyopathy cell injury model in Example 1 of the present invention, and the model selection of the LPS-induced time point in the subsequent experiments. Wherein, A: cell activity detection of each group; B+C: xCT protein expression level in each group of cells.

[0016] Figure 2 This is the construction of the cecal ligation and puncture model (CLP) of septic cardiomyopathy injury model in Example 2 of the present invention. Wherein, A: Schematic diagram of animal model grouping; B+C: CLP construction of each group.

[0017] Figure 3 The expression levels of the key disulfide death protein xCT at different time points were constructed for the CLP sepsis cardiomyopathy injury model according to Example 3 of the present invention.

[0018] Figure 4 This is the protective effect of 2-ME in Example 4 of the present invention on myocardial injury cells induced by LPS sepsis, and is a cell activity experimental detection.

[0019] Figure 5 The protective effect of 2-ME in Example 5 of the present invention on myocardial injury cells induced by LPS sepsis was detected by flow cytometry cell death experiment.

[0020] Figure 6 This is the fluorescence staining detection of 2-ME increasing the contractility of myocardial cells damaged by sepsis according to Example 6 of the present invention, wherein from left to right are actin-tracker red fluorescence staining, cell nucleus DAPI blue fluorescence staining, and actin-tracker+DAPI merge.

[0021] It should be noted that these drawings and textual descriptions are not intended to limit the conceptual scope of the present invention in any way, but are intended to illustrate the concept of the present invention for those skilled in the art by referring to specific embodiments. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. The following embodiments are used to illustrate the present invention but are not used to limit the scope of the present invention.

[0023] If the specific conditions are not specified in the specific implementation method, the conventional conditions or the conditions recommended by the manufacturer shall be followed. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially. The CCK-8 kit for cell activity experiment in the specific implementation method was purchased from Beyotime with the item number C0038; the Annexin V-FITC kit for cell death experiment was purchased from Beyotime with the item number C1062; the Actin-Tracker Red-Rhodamine kit for cytoskeletal protein staining experiment was purchased from Beyotime with the item number C2207S; the Lipopolysaccharides (LPS) from Escherichia coliO127:B8 reagent was purchased from Sigma with the item number L4516.

[0024] All experiments in the specific implementation were repeated at least 3 times independently, and the data were statistically analyzed using Graph Pad Prism 8.0, and the results were expressed as mean ± standard deviation. The t test was used for comparison between two groups; one-way analysis of variance was used for comparison between multiple groups, and Dunnett's test was used for multiple comparisons between two groups. P < 0.05 indicated that the difference was statistically significant.

[0025] Example 1

[0026] This example studies the optimal conditions for establishing a cell model of myocardial injury caused by sepsis:

[0027] 1. Cell and reagent preparation:

[0028] HL-1 mouse cardiomyocytes were cultured in high-glucose DMEM medium containing 10% fetal bovine serum, 100U / mL penicillin and 100mg / mL streptomycin. The cells were cultured in an incubator at 37°C, 20% oxygen and 5% carbon dioxide. The culture medium was changed every 48 hours. When the cell density was 70% to 80%, different concentrations of LPS purchased from Sigma with the catalog number L4516 were used to induce the cell model of sepsis myocardial injury. The administration time was 12 hours.

[0029] 2. Experimental groups:

[0030] Five LPS concentrations were selected: 0.5, 1, 2, 4, and 8 μg / ml, and two time periods were set: 12 and 24 hours.

[0031] 3. Cell activity The relative cell activity was detected by CCK8 (cell counting kit-8) experiment:

[0032] The cells were seeded in a 96-well plate, and 100uL of cell suspension was added to each well at a concentration of 5×10 4 / well, after pre-culturing the cells in the incubator for 24 hours, add different cell culture solutions containing different concentrations of LPS to each group of cell wells in the culture plate (set up 6 sub-wells for each group), and continue to culture in the cell incubator for 12 or 24 hours after adding drugs. Add 10μL of CCK-8 reagent to each well. After incubation for 2 hours, measure the absorbance at 450nm with an enzyme marker. Relative cell activity (%) = [(As-Ab) / (Ac-Ab)] ×100%. As: absorbance of the experimental group with LPS added; Ab: absorbance of the blank group; Ac: absorbance of the Ctrl group without LPS added.

[0033] 4. WB experiment to detect xCT protein expression:

[0034] RIPA lysis buffer containing Phenylmethanesulfonyl Fluoride (PMSF) (Coolaber, SL1020, Beijing, China) was used to lyse LPS-induced HL-1 cells in different groups to extract proteins. Proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Bio-Rad Laboratories, Inc., 5671034, Hercules, CA) and transferred to PVDF (Thermo Fisher Scientific Inc, 88520, Waltham, MA) membranes. After blocking, the PVDF membranes were incubated with primary antibodies at 4°C overnight. Then, the PVDF membranes were incubated with secondary antibodies at room temperature for 2 hours (1:3000, Abcam, ab6721, Cambridge, England). The primary antibodies used were xCT (Abcam, ab307601, Cambridge, England) and β-actin (Abcam, ab8226, Cambridge, England). The experiment was repeated three times.

[0035] Test results such as Figure 1 As shown, Figure 1Figure A in the middle shows the cell activity of each group of cells after treatment. When the LPS induction concentration is 1μg / ml, the CCK8 value is closest to 50%. This result shows that for the HL-1 mouse cardiomyocyte line, the LPS induction concentration of 1μg / ml is the optimal concentration for studying the cell injury model. Figure 2 B+C show the expression levels of xCT at different induction times when the LPS concentration was 1 μg / ml. It can be seen that compared with the Control group, the expression level of xCT was significantly upregulated at 12 hours and significantly downregulated at 24 hours. Because we want to discuss the pathogenic mechanism of xCT-induced disulfide death in septic cardiomyopathy, we chose the time period when xCT expression was significantly upregulated as the research object (n = 3, P < 0.01).

[0036] The results showed that LPS 1μg / ml for 12 hours was the optimal condition for LPS-induced myocardial injury in sepsis.

[0037] Example 2

[0038] This example studies the method for constructing an in vivo mouse injury model of septic cardiomyopathy:

[0039] 1. Animal and reagent preparation:

[0040] The cecal ligation and puncture (CLP) model is a widely used experimental model for studying sepsis. It induces systemic inflammatory response and multi-organ dysfunction by simulating bacterial peritonitis. The following is the experimental method for constructing the mouse sepsis CLP model in this implementation: 8-week-old male mice C57 strain were selected. The animals were adapted for one week and randomly divided into a control group and a model group. Mice were anesthetized with isoflurane or sodium pentobarbital. Ensure that the anesthesia is adequate by pinching the toe test reaction. The mouse was fixed with the abdomen facing up, the abdominal hair was removed, and it was disinfected with 75% alcohol and iodine. An incision of about 1-2 cm was made in the middle of the lower abdomen of the mouse to open the abdominal cavity. The cecum was found and its distal end was carefully separated from the mesentery of the large intestine. Sterile No. 4 silk thread was used to tightly ligate the distal 1 / 2 of the cecum. A sterile 21-gauge needle was used to puncture the ligation site and gently squeeze out a small amount of intestinal contents to simulate bacterial infection. The cecum was placed back into the abdominal cavity and the peritoneum and skin were sutured. An appropriate amount of saline was given according to the animal's weight to replenish the fluid loss during surgery. The animal was placed on a constant temperature heater to maintain body temperature to prevent hypothermic shock from causing death. The success of the model was evaluated by detecting the levels of inflammatory factors (such as TNF-α, IL-6), white blood cell counts, lymphocyte counts, and platelet counts in mice. In addition, the success of the septic cardiomyopathy model was evaluated by detecting cardiac function indicators through ultrasound.

[0041] 2. Experimental groups:

[0042] The cells were divided into three experimental groups at 12, 24, and 48 hours after CLP modeling.

[0043] 3. Small animal ultrasound cardiac function test:

[0044] First, anesthetize the mouse. Avertin or isoflurane can be used. The anesthetic dose is usually 0.5-0.75 mg / g body weight, and anesthesia is given by intraperitoneal injection or nasal inhalation. After anesthesia, use a depilatory or razor blade to remove the hair on the mouse's chest to facilitate contact with the ultrasound probe and reduce signal attenuation. Place the mouse on the operating table with its abdomen facing up, apply pure water (for conductivity) to the limbs and fix them to the corresponding position on the operating table with tape. Make sure that the mouse's head is slightly higher than the tail so that the apex and bottom of the heart are flush. Apply conductive glue about 2 cm thick on the mouse's chest to improve the conduction of ultrasound signals. Ultrasound detection: Long axis section: Place the ultrasound probe on the mouse's chest, the "notch" on the probe should face the animal's head, and rotate approximately 30° to 45° counterclockwise. You can obtain a B-mode echocardiogram of the long axis of the left ventricle, then turn on the M-mode ultrasound mode, adjust the sampling line to the correct position, and press the M-mode ultrasound mode again to obtain an M-mode echocardiogram of the long axis of the left ventricle. Short axis section: Turn the probe notch clockwise about 90° and adjust the probe to fit the mouse skin until a clear circular cardiogram with two papillary muscles is displayed. Repeat the above steps to obtain B-type and M-type echocardiograms. Measurement parameters: The following parameters are measured by echocardiography: Left ventricular ejection fraction (EF%): reflects the percentage of ventricular ejection and is an important indicator of cardiac function. Left ventricular short axis shortening rate (FS%): reflects the shortening function of myocardial fibers. Left ventricular end-diastolic diameter (LVID.d) and end-systolic diameter (LVID.s): used to evaluate the contractile and diastolic function of the heart. Data analysis: Analyze the acquired ultrasound images, record and calculate the above parameters to evaluate the cardiac function of mice.

[0045] Test results such as Figure 2 As shown in the figure, the cardiac function indexes of mice were significantly decreased after CLP modeling, and the septic cardiomyopathy model was successfully established (n = 3, ** indicates P < 0.01 compared with the Sham group)

[0046] Example 3

[0047] This example studies the optimal conditions for constructing a septic cardiomyopathy injury model in mice:

[0048] 1. Animal and reagent preparation refer to Example 2.

[0049] 2. Experimental grouping refers to Example 2.

[0050] 3. WB experiment to detect xCT protein expression:

[0051] RIPA lysis buffer containing Phenylmethanesulfonyl Fluoride (PMSF) (Coolaber, SL1020, Beijing, China) was used to lyse LPS-induced HL-1 cells in different groups to extract proteins. Proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Bio-Rad Laboratories, Inc., 5671034, Hercules, CA) and transferred to PVDF (Thermo Fisher Scientific Inc, 88520, Waltham, MA) membranes. After blocking, the PVDF membranes were incubated with primary antibodies at 4°C overnight. Then, the PVDF membranes were incubated with secondary antibodies at room temperature for 2 hours (1:3000, Abcam, ab6721, Cambridge, England). The primary antibodies used were xCT (Abcam, ab307601, Cambridge, England) and β-actin (Abcam, ab8226, Cambridge, England). The experiment was repeated three times.

[0052] Test results such as Figure 3 As shown, the expression levels of xCT in the myocardium of mice at different time points after CLP modeling are shown. It can be seen that compared with the Sham group, the expression level of xCT was significantly upregulated at 12 hours and significantly downregulated at 24 hours, and the trend was consistent with the in vitro cell model. We selected the time period when xCT expression was significantly upregulated as the research object (n = 3, P < 0.01).

[0053] This indicates that CLP modeling for 12 hours is the optimal condition for sepsis-induced myocardial injury.

[0054] Example 4

[0055] This example studies whether 2-ME can restore the activity of myocardial cells damaged by sepsis:

[0056] 1. Cell and reagent preparation:

[0057] HL-1 mouse cardiomyocytes were cultured in high-glucose DMEM medium containing 10% fetal bovine serum, 100U / mL penicillin and 100mg / mL streptomycin. They were cultured in an incubator at 37°C, 20% oxygen and 5% carbon dioxide. The culture medium was changed every 48 hours, and a cell model of septic myocardial injury was induced using LPS (1μg / ml) purchased from Sigma with a catalog number of L4516 when the cell density was 70% to 80%. The administration time was 12 hours. 2-ME was purchased from Merck with a catalog number of M6250, prepared with 1μg / ml LPS culture medium, with a concentration of 20μM, and the administration time was 12 hours.

[0058] 2. Experimental groups:

[0059] Ctrl: HL-1 mouse cardiomyocytes were routinely cultured;

[0060] LPS: HL-1 mouse cardiomyocytes were induced by administration of 1 μg / ml LPS for 12 h;

[0061] LPS+2-ME: HL-1 mouse cardiomyocytes were induced by LPS at 1 μg / ml for 12 hours, and 2-ME was added at a final concentration of 20 μM.

[0062] 3. Detect relative cell activity through CCK8 assay:

[0063] The cells were seeded in a 96-well plate, and 100uL of cell suspension was added to each well at a concentration of 5×10 4 / well, after pre-culturing cells in the incubator for 24 hours, add drugs according to different cell groups (set 6 sub-wells for each group), and continue to culture in the cell incubator for 12 hours after adding drugs. Add 10μL of CCK-8 reagent to each well. After incubation for 2 hours, measure the absorbance at 450nm with an enzyme marker. Relative cell activity (%) = [(As-Ab) / (Ac-Ab)] ×100%. As: absorbance of the experimental group with LPS; Ab: absorbance of the blank group; Ac: absorbance of the Ctrl group without LPS.

[0064] Test results such as Figure 4 As shown, in the LPS HL-1 mouse cardiomyocyte sepsis injury model, administration of the disulfide bond stress reducing agent 2ME (20 μM) can restore the activity of mouse cardiomyocytes under sepsis injury.

[0065] Example 5

[0066] This example studies whether 2-ME can reduce myocardial cell death caused by sepsis:

[0067] 1. Cells and reagents were prepared as described in Example 4.

[0068] 2. Experimental grouping refers to Example 4.

[0069] 3. Flow cytometry to detect cell death:

[0070] Aspirate the cell culture medium into a suitable centrifuge tube, wash the adherent cells once with PBS, and add an appropriate amount of trypsin cell digestion solution to digest the cells. Incubate at room temperature until the adherent cells can be blown off by gently blowing, and then aspirate the trypsin cell digestion solution. Avoid excessive trypsin digestion. Note: For adherent cells, the trypsin digestion step is critical. If the trypsin digestion time is too short, the cells need to be blown hard to fall off, which is easy to cause cell membrane damage, resulting in false positive cell necrosis; if the digestion time is too long, it is also easy to cause cell membrane damage and false positive cell necrosis, and even affect the binding of phosphatidylserine on the cell membrane with Annexin V-FITC, thereby interfering with the detection of cell apoptosis. At the same time, the trypsin cell digestion solution should be free of EDTA as much as possible, because EDTA may affect the binding of Annexin V with phosphatidylserine. Add the collected cell culture medium, gently blow the cells off, transfer to a centrifuge tube, centrifuge at 1000g for 5 minutes, discard the supernatant, collect the cells, gently resuspend the cells with PBS and count them. Take 10,000 resuspended cells, centrifuge at 1000g for 5 minutes, discard the supernatant, add 5µl Annexin V-FITC, 10µl propidium iodide staining solution, and mix gently. Incubate at room temperature (20-25ºC) in the dark for 20 minutes. Detection can be performed on the flow cytometer immediately. Annexin V-FITC is green fluorescence, and propidium iodide (PI) is red fluorescence. PI+ is judged as dead cells.

[0071] Test results such as Figure 5 As shown, the cell death rate in the LPS-induced sepsis cardiomyocyte injury model was significantly increased (n=3, P<0.05), while 2-ME could reduce the cell death rate of cardiomyocytes under sepsis injury.

[0072] Example 6

[0073] This example studies the phenomenon that 2-ME can inhibit the contraction of septic cardiomyopathy cells:

[0074] 1. Cells and reagents were prepared as described in Example 4.

[0075] 2. Experimental grouping refers to Example 4.

[0076] 3. Cytoskeleton protein actin staining:

[0077] Wash the cells twice with PBS; fix the cells with 3.7% formaldehyde solution at room temperature for 20 minutes, and then wash them with 0.1% Triton X-100 in PBS for 3 times, 5 minutes each time; dilute Actin-Tracker Red with PBS containing 5% BSA and 0.1% Triton X-100 at a ratio of 1:100 as the staining working solution; stain the collected cells with the staining working solution and incubate them at room temperature in the dark for 60 minutes. Observe and take pictures with a fluorescence microscope. Actin emits red fluorescence with a maximum excitation wavelength of 540nm and a maximum emission wavelength of 565nm.

[0078] Test results such as Figure 6 As shown, in the LPS-induced cardiomyocyte injury model, cells contracted significantly, and this phenomenon was significantly alleviated after the addition of 2ME inhibitor.

[0079] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. 2-ME is used in the preparation of drugs for preventing or treating septic cardiomyopathy.

2. The use according to claim 1, characterized in that: The dosage of 2-ME is 20umol / L.

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