WO2021073249A1 - USE OF β-NMN IN PREPARATION OF DRUG FOR TREATING AND PREVENTING SEPSIS-INDUCED ORGAN DAMAGE - Google Patents

Abstract

Disclosed is the use of β-nicotinamide mononucleotide in the preparation of a drug for treating and/or preventing sepsis-induced organ damage. The new use of β-nicotinamide mononucleotide has a therapeutic effect and a preventive effect on sepsis-induced multiple organ damage, and β-nicotinamide mononucleotide is easily soluble in water, is widespread and highly operable, and has various administration routes.

Description

Application of β-NMN in the preparation of drugs for the treatment and prevention of sepsis organ damage Technical field

The invention relates to the field of biomedicine, in particular to the application of β-NMN in the preparation of a medicine for the treatment and prevention of septic organ damage.

Background technique

Sepsis (sepsis) is a life-threatening multiple organ dysfunction caused by the imbalance of the body response after infection, which can develop into septic shock and multiple organ dysfunction syndrome (MODS), which is clinical One of the leading causes of death in critically ill patients. According to reports, there are nearly 18 million sepsis patients in the world each year, and the number of sepsis patients in my country is estimated to be more than 5 million each year, and the number of patients is increasing year by year at a rate of nearly 10%, causing a serious medical burden. The research data of domestic and foreign scholars shows that despite the continuous progress of "clustered" treatment strategies such as capacity assessment and resuscitation, early anti-infection and organ support, the mortality rate of patients remains high. So far, there are no effective measures to prevent and treat sepsis. Therefore, it is urgent to find new targets and new treatment measures for the prevention and treatment of sepsis.

So far, there is no clinically effective treatment for sepsis. At present, supportive therapy is the main clinical practice, which is briefly described as follows:

1. Fluid resuscitation: Most patients with sepsis will have circulatory failure in the early stage. It is determined that patients with sepsis have persistent hypotension. Before being admitted to the intensive care unit, the initial resuscitation and volume expansion treatment will be carried out through intravenous rehydration. However, in patients with sepsis, the diastolic function of the heart is limited, and a large amount of crystalloid infusion within a short period of time may exceed the compensatory capacity of the patient’s heart. This will not only not increase cardiac output, but will cause pulmonary edema and high center. Serious hemodynamic consequences including liver and kidney damage caused by venous pressure. Therefore, this also puts forward higher requirements for clinicians in the evaluation of fluid type, measurement, and course of treatment.

2. Application of vasoactive drugs: In order to make the patient's average arterial pressure greater than or equal to 65mmHg, vasopressor drugs are needed. Generally, norepinephrine is the first choice, and vasopressin can be used to increase the average arterial pressure value. However, the actual application, dosage and choice of drugs for patients with septic shock are also controversial. In addition to norepinephrine and vasopressin, angiotensin II has gradually become more prominent in the role of vasoactive drugs. Studies have found that angiotensin II can effectively increase the arteries of patients with vasodilatory shock whose conventional vasoactive drugs are ineffective. blood pressure. As for the timing of the application of vasoactive drugs and the appropriate reduction or even discontinuation, there is still no unified conclusion.

3. The use of antibiotics: Earlier studies have found that for every hour of delay in applying antibiotics in patients with septic shock, the fatality rate is estimated to increase by 7.6%. Subsequent studies did not find a correlation between the time from emergency triage to antibiotic application 6 hours before resuscitation treatment and the mortality rate during hospitalization, but delaying the application of antibiotics to septic shock significantly increased the risk of death. Recent clinical studies have found that there is no statistically significant difference in the 28-day or 90-day mortality of sepsis patients with antibiotics in ambulances compared with antibiotics after admission. The current evidence does not support that all patients with sepsis should use antibiotics within a strict time window, and the abuse of antibiotics can also cause an increase in drug resistance.

4. Other adjuvant treatment methods: patients with severe sepsis also need to use high-flow nasal catheters or mask oxygen to protect ventilation to treat respiratory distress and hypoxemia; when the patient's blood sugar level continuously exceeds 180mg/dl, then Insulin is needed, but the blood sugar level of capillaries detected by bedside indicators cannot fully reflect the blood sugar of the patient's plasma and arteries.

In summary, clinically, fluid resuscitation, the use of vasoactive drugs and anti-infective drugs have always been the core strategy of the campaign to save sepsis, but there is a lack of specific treatment measures. Therefore, it is particularly urgent to continue pre-clinical basic research to provide effective and specific prevention and treatment programs.

Nicotinamide Mononucleotide (NMN) is a nucleotide derived from ribose and nicotinamide. It can be synthesized in the body through two pathways: one is catalyzed by nicotinamide via nicotinamide phosphoribosyl transferase (NAMPT) Synthesis; the other way is by Nicotinamide riboside (Nicotinamide riboside) catalyzed synthesis by nicotinamide riboside kinase. β-Nicotinamide Mononucleotide (β-Nicotinamide Mononucleotide, β-NMN) is the active form of nicotinamide mononucleotide, which acts as Nicotinamide adenine dinucleotide (NAD ⁺ ; also known as coenzyme) The function of the precursor of I) is mainly realized by NAD ⁺ . NAD ⁺ is a coenzyme that transfers protons. It is an indispensable coenzyme in glycolysis, gluconeogenesis, tricarboxylic acid cycle, and respiratory chain. It plays an important role in providing energy to cells, repairing DNA, and anti-aging. The role of substitution. Therefore, it is particularly important to ^{maintain the content of NAD + in the cell.} Although there are many ways to synthesize NAD ⁺ in the body, studies have shown that β-nicotinamide mononucleotide is produced by the catalysis of Nicotinamide nucleotide adenylytransferase 1-3 (NMNAT1-3) NAD ⁺ accounts for 85% of the total body's NAD ⁺ and is the most important way to ^{maintain the NAD+ content in the body.} Interestingly, studies have reported that the NAD ⁺ content in the body significantly decreases under sepsis conditions, and it may be closely related to the continued deterioration of the main organs in sepsis. Recent studies by the inventor’s team have confirmed that the application of nicotinamide riboside to increase the NAD ⁺ content can effectively reduce the damage to the main organs of sepsis, but it requires preventive administration before the occurrence of sepsis, such as administration of cigarettes after the occurrence of sepsis. Amide riboside has no protective effect. The reason is that sepsis conditions severely reduce the expression of nicotinamide riboside kinase, which causes the nicotinamide riboside to be unable to be converted into β-nicotinamide mononucleotide in the body (Hong G, et al. Free Radiac Bio Med 2018; 123: 125-137). Therefore, nicotinamide riboside cannot be used to treat major organ damage in sepsis.

Summary of the invention

In order to solve the above technical problems, the purpose of the present invention is to provide an application of β-NMN in the preparation of drugs for the treatment and prevention of septic organ damage. The present invention discloses a new type of treatment and/or prevention of septic organ damage. medicine.

The technical scheme of the present invention is as follows:

The invention discloses the application of β-nicotinamide mononucleotide (β-NMN) in the preparation of medicines for the treatment and/or prevention of organ damage caused by sepsis.

Further, organ damage includes one or more of heart damage, lung damage, liver damage, and kidney damage.

Further, the drug is administered by injection or oral administration.

Further, injection administration includes intravenous and/or intraperitoneal injection administration.

Further, the administration dose of the drug is 300-1000 mg/kg. Preferably, the dosage of the drug is 500 mg/kg.

Further, the medicine is used to increase the ^{reduction of coenzyme I (NAD +} ) content caused by sepsis.

Further, the medicine is used to reduce the activity of myeloperoxidase and the content of active oxygen and malondialdehyde in organs caused by sepsis.

With the above solution, the present invention has at least the following advantages:

1. The present invention discloses the new use of β-nicotinamide mononucleotide, which has therapeutic, preventive and protective effects on multiple organ damage caused by sepsis, thereby providing new treatment methods and approaches for diseases in related fields. Before or after the onset of sepsis, a single administration of β-nicotinamide mononucleotide can increase the ^{level of NAD +} in the body, improve the heart function of septic mice, and reduce the level of oxidative stress in sepsis mice. Reduce the damage of heart, liver, lung, kidney and other organs.

2. The preparation method of β-nicotinamide mononucleotide is simple: β-nicotinamide mononucleotide is easy to dissolve in water, no other special solvents are required, and it is very popular and maneuverable.

3. The route of administration can be diversified: In the present invention, both intravenous injection and intraperitoneal injection are effective. In addition, it can also be administered orally.

The above description is only an overview of the technical solution of the present invention. In order to understand the technical means of the present invention more clearly and implement it in accordance with the content of the description, the preferred embodiments of the present invention are described below with detailed drawings.

Description of the drawings

Figure 1 is the detection result of heart function, oxidative stress and inflammatory response in Example 2 of the present invention;

Figure 2 is the detection results of lung histopathology, oxidative stress, inflammatory response and cell apoptosis in Example 2 of the present invention;

Fig. 3 is the detection result of oxidative stress, cell damage and inflammatory reaction of liver tissue in Example 2 of the present invention;

Figure 4 is the detection results of oxidative stress, cell damage and inflammatory reaction in kidney tissue in Example 2 of the present invention;

Figure 5 is the detection results of heart function, oxidative stress and inflammatory response in Example 3 of the present invention;

Figure 6 is the detection results of lung tissue oxidative stress, inflammatory response and cell apoptosis in Example 3 of the present invention;

Fig. 7 is the detection result of oxidative stress, cell damage and inflammatory reaction of liver tissue in Example 3 of the present invention;

Fig. 8 shows the detection results of oxidative stress, cell damage and inflammatory reaction in kidney tissue in Example 3 of the present invention.

Detailed ways

The specific implementation of the present invention will be described in further detail below in conjunction with examples. The following examples are used to illustrate the present invention, but not to limit the scope of the present invention.

The present invention mainly uses the following experimental methods to detect representative biochemical parameters and damage indicators of multiple organs in septic mice, which are briefly described as follows:

1. Ultrasound evaluation of cardiac function: 1% isoflurane anesthetize mice, use high-resolution cardiac ultrasound probe (35-MHz linear array transducer attached to Vevo 2100 ultrasonic system) to obtain a ventricular section at the level of the papillary muscle of the mitral valve. The M-mode image measures the thickness of the anterior and posterior walls of the ventricle, the diameter of the ventricular cavity and the movement of the ventricular wall, and then analyzes the systolic function: short axis shortening rate (FS%) and left ventricular ejection fraction (EF%). In the apical four-chamber view, pulse Doppler tissue echocardiography was used to analyze the diastolic function: peak atrial blood flow velocity A, peak early diastolic blood flow velocity E, and their ratio (E/A).

2. Serological testing: Collect mouse peripheral serum, and use commercial kits to detect alanine aminotransferase (Alamine aminotransferase, also known as GPT) and aspartate aminotransferase (AST also known as GOT) that reflect liver function indicators; reaction Creatinine (Cr) and blood urea nitrogen (BUN), which are indicators of renal function.

3. Detection of tissue reactive oxygen species: Grind mouse heart, lung, liver and kidney tissues, add horseradish peroxidase and Amplex Red to detect the amount of reactive oxygen species produced in each tissue. The final form of reactive oxygen species in tissue cells is hydrogen peroxide. The product of hydrogen peroxide catalyzed by horseradish peroxidase can be labeled with a fluorescent signal carried by Amplex Red. The strength of the fluorescence signal represents the production of reactive oxygen species in the tissue.

4. Malondialdehyde (MDA) detection: The detection method is based on the color reaction of MDA and Thiobarbituric acid (Thiobarbituric acid, TBA) to produce a red product. MDA is a natural product produced by organisms after lipid oxidation. When the cells of the body undergo oxidative stress, lipid oxidation also occurs. Some fatty acids will be decomposed into a series of complex compounds after oxidation, including MDA. The level of lipid oxidation in the body can be reflected by detecting the content of MDA, so the measurement of MDA is widely used as an indicator of lipid oxidation. At higher temperatures in an acidic environment, MDA can react with TBA to form a red MDA-TBA adduct. The latter has a maximum absorption at 535nm, so it can be detected by colorimetry. The specific reaction principle is as follows:

5. Detection of protein carbonyl groups: during oxidative stress, the effects of free radicals on proteins include protein peptide chain scission, cross-linking and polymerization between protein molecules, oxidative deamination of protein amino acids, and free radicals attacking protein reducing groups, The malondialdehyde produced by oxidative cleavage of lipids is cross-linked with molecules produced by amino groups on proteins. At present, there are two main detection indicators for protein oxidative damage, namely protein carbonyl production and nitrotyrosine production. In the present invention, the protein carbonyl detection kit of Sigma-Aldrich is used to form a stable dinitrophenylhydrazine complex by the interaction between the protein carbonyl and 2,4-dinitrophenylhydrazine (DNPH), the latter can be detected at 375nm Maximum light absorption.

6. Detection of cell apoptosis: The commonly used detection methods for cell apoptosis mainly include: caspas-3 activity determination, caspase-3 cleavage fragment determination and TUNEL in situ staining. Among them, caspase-3 activity determination and TUNEL in situ staining are performed using commercial kits, and caspase-3 cleavage fragments are detected by western blot using related antibodies.

7. Detection of pulmonary capillary permeability: Evans blue staining method is mainly used in the present invention to evaluate the permeability of pulmonary capillary vessels. 30 minutes before euthanasia, the mice were injected with 0.4% Evans Blue solution (50 mg/kg) through the tail vein. After lavage with 10 ml of normal saline, the mouse lung tissue was weighed and homogenized with normal saline. Mix the tissue lysate with 2ml of formamide, incubate at 60°C for 16 hours and then centrifuge at 20000g at 4°C for 5 minutes. Aspirate the supernatant to determine the concentration of Evans Blue by spectrophotometry (620, 740nm), and evaluate the permeability of the capillaries by the leakage rate of Evans Blue in the lung tissue.

8. Detection of myeloperoxidase (myeloperoxidase, MPO): Myeloperoxidase is a heme protease containing heme prosthetic group secreted by neutrophils, monocytes and macrophages in certain tissues. It is a member of the heme peroxidase superfamily. Myeloperoxidase is a function and activation mark of neutrophils, and its level and activity changes represent the function and activity status of neutrophil polymorphonuclear leukocytes. In the present invention, a commercial kit is used to reflect the level of enzyme activity by detecting the content of the myeloperoxidase catalytic substrate.

9. Histopathological analysis: In the present invention, Hematoxylin-Eosin (HE) staining method is mainly used for pathological analysis of mouse lung tissue. After the mice were euthanized, the right lobe of the lung was collected, and an appropriate volume of 10% formalin tissue fixative was added. After being allowed to stand at room temperature for 24 hours, it was dehydrated by alcohol and embedded in paraffin. A paraffin section with a thickness of 0.5 mm was cut with a paraffin microtome, and hematoxylin-eosin staining was performed through deparaffinization, hydration, nuclear and cytoplasmic staining, and decolorization, and then the pathological changes of lung tissue structure were observed under a microscope.

Example 1: Preparation of mouse sepsis model

The following examples of the present invention use the method of intraperitoneal injection of stool to prepare a sepsis mouse model, and the specific method steps are as follows:

1. Mice: 7-8 weeks old C57 male mice (body weight around 25 grams) are placed in the animal turnover room for later use. Randomly select several mice as fecal donor mice for future use. Then the remaining mice were randomly divided into four groups: the control group, the control group plus β-nicotinamide mononucleotide group, the implanted stool group and the implanted stool plus β-nicotinamide mononucleotide group (the number of each group was greater than 5). only).

2. Stool collection: After euthanasia, the donor mice were dissected, and the cecum was collected and weighed. Each 75 mg of feces was dissolved in 1 ml of normal saline (the implanted feces volume was 50 ml/kg body weight), and mixed thoroughly. 4 Let stand for 24 hours, set aside. For example, to prepare a sepsis model for 10 mice weighing 25 grams, 938 mg of feces should be collected and dissolved in 12.5 ml of normal saline.

3. Implant feces to prepare a sepsis model: take out the prepared fecal suspension, and implant feces into the abdominal cavity of experimental mice by intraperitoneal injection at a ratio of 50 ml/kg body weight. Subsequently, 1 ml of normal saline containing 4 micrograms of buprenorphine was subcutaneously injected into the mice to replenish electrolytes and relieve pain in the mice. The end of the experiment was 6 hours later, and the damage of the heart, liver, and kidney of mice in different experimental groups was analyzed. The control group and the control group plus β-nicotinamide mononucleotide group did not undergo any operation, the stool implantation group and the stool implantation plus β-nicotinamide mononucleotide group were first prepared according to the above method to prepare the sepsis model.

Example 2: β-nicotinamide mononucleotide prevents multiple organ damage in septic mice

In the present invention, the administration method of intraperitoneal injection is adopted, and the specific steps are as follows:

1. Prepare β-nicotinamide mononucleotide solution: weigh the β-nicotinamide mononucleotide required by the experimental mice at a ratio of 500 mg/kg body weight, and dissolve it in an appropriate amount of normal saline (each mouse The injection volume is 100 microliters). For example, when preparing the β-nicotinamide mononucleotide injection required for 10 mice weighing 25 grams, the weighed 125 mg of β-nicotinamide mononucleotide should be dissolved in 1 ml of normal saline.

2. Intraperitoneal injection of β-nicotinamide mononucleotide: For the implanted stool plus β-nicotinamide mononucleotide group constructed in Example 1, 100 microliters were injected into the mouse's abdominal cavity while the experimental mice were implanted in the stool The prepared β-nicotinamide mononucleotide solution, for the control group of Example 1 plus β-nicotinamide mononucleotide group, the mice were injected with the same amount of β-nicotinamide mononucleotide. The control group and the stool implantation group were injected with the same amount of normal saline.

The heart function of the four groups of mice was measured by ultra-high resolution echocardiography after implantation of feces for 6 hours. After euthanasia, peripheral serum and heart, liver, lung and kidney tissue samples were collected from all mice for analysis of different tissues.

The mice were anesthetized with 1-2% isoflurane, and the contractile function of the mouse heart was detected by ultra-high-resolution echocardiography. The results are shown in Figure 1. In Figure 1, Vehicle represents control mice; Feces represents sepsis In the mouse group, Saline represents normal saline; NMN represents the application of β-nicotinamide mononucleotide at the onset of sepsis. Fractional Shortening (%) is the short-axis shortening rate, which represents cardiac contractility; MPO represents myeloperoxidase activity; ROS represents the production of reactive oxygen species; MDA represents the content of malondialdehyde. The data in Figure 1 is the mean ± standard deviation, using one-way analysis of variance, n=5, *P＜0.05vs Vehicle (Saline),

vs Feces(Saline). In Figure 1, the cardiac contractile function of septic mice was significantly weaker than that of control mice, and β-nicotinamide mononucleotide improved the cardiac function of septic mice (Figure 1A). Grinding mouse myocardial tissues, using a commercial kit to detect the activity of myeloperoxidase and the production of reactive oxygen species and malondialdehyde in myocardial tissues. The results are shown in Figure 1B-D. Sepsis leads to myocardial tissue marrow. Increased peroxidase activity and excessive production of reactive oxygen species and malondialdehyde, β-nicotinamide mononucleotide significantly reduced myeloperoxidase activity and reactive oxygen species and malondialdehyde in myocardial tissue of septic mice content. The results show that β-nicotinamide mononucleotide can improve the heart function of septic mice.

Figure 2 shows the relevant test results of lung tissue. In the figure, Caspase-3 activity represents the level of apoptosis. The other letters and symbols have the same meaning as in Figure 1. The lung tissue is embedded in formalin, dehydrated and embedded in paraffin. After sectioning, hematoxylin-eosin staining was performed. The representative picture is shown in 2A: the lung tissue of septic mice showed inflammatory reactions such as alveolar wall thickening and cell nucleus aggregation. β-nicotinamide mononucleotide effectively improved sepsis Inflammatory damage to lung tissue caused by disease. This result is also consistent with the detection results of myeloperoxidase activity representing inflammatory injury, reactive oxygen species and malondialdehyde representing the level of oxidative stress, and apoptosis (Figure 2B-D). Sepsis induces apoptosis of lung tissue cells, and β-nicotinamide mononucleotide reduces the level of apoptosis in lung tissue cells of sepsis mice (Figure 2E). In summary, β-nicotinamide mononucleotide can alleviate lung tissue lesions in septic mice.

Figure 3 shows the relevant test results of liver tissue. In the figure, Protein carbonyl represents the content of protein carbonyl group, AST represents aspartate aminotransferase that causes liver cell damage, and other letters and symbols have the same meaning as Figure 1. Protein in liver tissue of septic mice The content of carbonyl and malondialdehyde and the activity of myeloperoxidase were significantly higher than those of control mice, suggesting that the liver tissue of sepsis mice had oxidative stress and inflammatory damage. Similarly, β-nicotinamide mono Nucleotides can significantly improve the level of oxidative stress and inflammatory damage in liver tissue caused by sepsis (Figure 3A, B and D). In addition, aspartate aminotransferase, which represents liver cell damage, was significantly increased in the peripheral serum of septic mice, and β-nicotinamide mononucleotide could reduce the expression of aspartate aminotransferase in septic mice (Figure 3C). In summary, β-nicotinamide mononucleotide can improve liver tissue lesions in septic mice.

Figure 4 shows the related test results of kidney tissue. In the figure, BUN stands for urea nitrogen that reflects kidney cell damage. Other letters and symbols have the same meaning as Figures 1 and 3. The protein carbonyl and malondialdehyde in the kidney tissue of septic mice The content and the activity of myeloperoxidase were significantly higher than those of control mice, suggesting that oxidative stress and inflammatory damage occurred in the kidney tissue of sepsis mice. Similarly, β-nicotinamide mononucleotide can be significantly Improve the level of oxidative stress and inflammatory damage in the kidney tissue caused by sepsis (Figure 4A, B and D). The increase in serum urea nitrogen content indicates renal insufficiency in septic mice. After the application of β-nicotinamide mononucleotide, the urea nitrogen content is down-regulated to protect renal function (Figure 4C). In summary, β-nicotinamide mononucleotide can improve renal tissue lesions in septic mice.

Example 3: β-nicotinamide mononucleotide treatment of multiple organ damage in septic mice

In the present invention, the administration method of intraperitoneal injection is adopted, and the specific steps are as follows:

1. Prepare β-nicotinamide mononucleotide solution: weigh the β-nicotinamide mononucleotide required by the experimental mice at a ratio of 500 mg/kg body weight, and dissolve it in an appropriate amount of normal saline (each mouse The injection volume is 100 microliters). For example, when preparing the β-nicotinamide mononucleotide injection required for 10 mice weighing 25 grams, the weighed 125 mg of β-nicotinamide mononucleotide should be dissolved in 1 ml of physiological saline.

2. Intravenous injection of β-nicotinamide mononucleotide: For the implanted feces plus β-nicotinamide mononucleotide group constructed in Example 1, one hour after the fecal implantation of the experimental mice, the mice were injected with 100 into the tail vein Microliters of the prepared β-nicotinamide mononucleotide solution, for the control group of Example 1 plus β-nicotinamide mononucleotide group, mice were injected with the same amount of β-nicotinamide mononucleotide. The control group and the stool implantation group were injected with the same amount of normal saline.

The heart function of the four groups of mice was measured by ultra-high resolution echocardiography after implantation of feces for 6 hours. After euthanasia, peripheral serum and heart, liver, lung and kidney tissue samples were collected from all mice for analysis of different tissues.

The mice were anesthetized with 1-2% isoflurane, and the contractile function of the mouse heart was detected by ultra-high resolution echocardiography. The results are shown in Figure 5. In the figure, NMN-1H represents the application of β- when sepsis is onset for 1 hour. Nicotinamide mononucleotide, other letters and symbols have the same meaning as above. The cardiac contractile function of sepsis mice was significantly weaker than that of control mice. One hour after the onset of sepsis, β-nicotinamide mononucleotide was used to improve the cardiac function of sepsis mice (Figure 5A). The mouse myocardial tissue was ground, and a commercial kit was used to detect the activity of myeloperoxidase and the production of reactive oxygen species and malondialdehyde in the myocardial tissue. The results are shown in Figure 5B-D. Sepsis leads to myocardial tissue marrow. Increased peroxidase activity and excessive production of reactive oxygen species and malondialdehyde. Application of β-nicotinamide mononucleotide 1 hour after the onset of sepsis significantly reduced myeloperoxidase activity in myocardial tissue of septic mice And the content of active oxygen and malondialdehyde. In summary, the application of β-nicotinamide mononucleotide one hour after the onset of sepsis can improve the heart function of septic mice.

Figure 6 illustrates the relevant test results of lung tissue. In the figure, NMN-1H represents the application of β-nicotinamide mononucleotide at the onset of sepsis one hour, and other letters and symbols have the same meaning as above. Formalin-embedded lung tissues were dehydrated and paraffin-embedded in sections and then stained with hematoxylin and eosin. A representative picture is shown in 6A: The lung tissue of septic mice showed inflammation such as alveolar wall thickening and cell nucleus aggregation. In response, the application of β-nicotinamide mononucleotide 1 hour after the onset of disease can effectively improve the inflammatory damage of lung tissue caused by sepsis. This result is also consistent with the detection results of myeloperoxidase activity representing inflammatory damage, reactive oxygen species and malondialdehyde representing the level of oxidative stress, and cell apoptosis (Figure 6B-D). Sepsis induces apoptosis of lung tissue cells, and β-nicotinamide mononucleotide is applied to reduce the apoptosis level of lung tissue cells in sepsis mice 1 hour after the onset (Figure 6C). In summary, one hour after the onset of sepsis, β-nicotinamide mononucleotide can be used to reduce lung tissue lesions in septic mice.

Figure 7 shows the results of the related tests of liver tissue. In the figure, NMN-1H represents the application of β-nicotinamide mononucleotide at the onset of sepsis one hour, and other letters and symbols have the same meaning as above. The content of protein carbonyl, malondialdehyde and the activity of myeloperoxidase in the liver tissues of sepsis mice were significantly higher than those of control mice, suggesting that the liver tissues of sepsis mice had oxidative stress and inflammatory damage Similarly, the application of β-nicotinamide mononucleotide 1 hour after the onset of onset can significantly improve the level of oxidative stress and inflammatory damage in liver tissue caused by sepsis (Figure 7A, B and D). In addition, aspartate aminotransferase, which represents liver cell damage, was significantly increased in the peripheral serum of septic mice, and the application of β-nicotinamide mononucleotide can reduce the expression of peripheral serum aspartate aminotransferase in septic mice ( Figure 7C). In summary, the application of β-nicotinamide mononucleotide one hour after the onset of sepsis can improve liver tissue lesions in septic mice.

Figure 8 illustrates the related test results of kidney tissue. In the figure, NMN-1H represents the application of β-nicotinamide mononucleotide at the onset of sepsis one hour, and other letters and symbols have the same meaning as above. The content of protein carbonyl and malondialdehyde and the activity of myeloperoxidase in the kidney tissue of sepsis mice were significantly higher than those of control mice, suggesting that the kidney tissue of sepsis mice had oxidative stress and inflammatory damage Similarly, the application of β-nicotinamide mononucleotide 1 hour after the onset of disease can significantly improve the level of oxidative stress and inflammatory damage in the kidney tissue caused by sepsis (Figure 8A, B and D). The increase in peripheral serum urea nitrogen content indicates renal insufficiency in septic mice. β-nicotinamide mononucleotide was used to down-regulate the urea nitrogen content and protect renal function one hour after the onset of disease (Figure 8C). In summary, β-nicotinamide mononucleotide can be used to improve renal tissue lesions in septic mice one hour after the onset of sepsis.

The present invention proves through preliminary in vivo experiments that the intraperitoneal injection of β-nicotinamide mononucleotide at the same time or after the preparation of the mouse sepsis model effectively improves the damage of the main organs of the mouse sepsis, and proves that the β-nicotinamide mononuclear The preventive and therapeutic protective effects of Glycolic acid on multiple organs of septic mice indicate that β-nicotinamide mononucleotide can be used to prepare drugs for the treatment and prevention of septic organ damage, thereby providing a good basis for diseases in related fields. New treatment methods and approaches.

The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. It should be pointed out that for those of ordinary skill in the art, several improvements can be made without departing from the technical principles of the present invention. And variants, these improvements and variants should also be regarded as the protection scope of the present invention.

Claims (6)

1. Application of β-nicotinamide mononucleotide in the preparation of drugs for the treatment and/or prevention of organ damage caused by sepsis.
2. The application according to claim 1, wherein the organ damage includes one or more of heart damage, lung damage, liver damage and kidney damage.
3. The application according to claim 1, wherein the drug is administered by injection or oral administration.
4. The application according to claim 3, wherein the injection administration includes intravenous and/or intraperitoneal injection administration.
5. The application according to claim 3, characterized in that: the dosage of the drug is 300-1000 mg/kg.
6. The application according to claim 1, characterized in that: the medicine is used to increase the reduction of coenzyme I content caused by sepsis.

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