Application of nitro fatty acid in preparation of product for predicting, preventing and improving adverse cardiovascular events of acute coronary syndrome patients

Abstract

The invention discloses an application of nitro fatty acid in preparation of a product for predicting, preventing and improving adverse cardiovascular events of patients with acute coronary syndrome. The invention provides application of nitro fatty acid in preparation of products for preventing and improving adverse cardiovascular events of ACS patients. According to the application, for the ACS patient, when the concentration of the 12-nitro conjugated linoleic acid in the body is larger than 0.196 ng / ml and the concentration of the 9-nitro conjugated linoleic acid is larger than 0.296 ng / ml, it is judged that the risk that the ACS patient is re-hospitalized and dead due to non-fatal myocardial infarction / unstable angina pectoris / blood supply reconstruction is low. By detecting the content of nitro fatty acid in a biological sample and supplementing nitro fatty acid, the occurrence rate of adverse events can be greatly reduced. Meanwhile, the invention further provides a detection method of nitro fatty acid, and the accuracy of biological sample detection is improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of nitro fatty acids in the preparation of products for predicting, preventing and improving adverse cardiovascular events in patients with acute coronary syndrome. Background Art

[0002] Acute coronary syndrome (ACS) is an acute adverse syndrome of the heart caused by a sudden reduction in myocardial blood supply. ACS includes ST-segment elevation myocardial infarction (STEMI), non-ST-segment elevation myocardial infarction (NSTEMI) or unstable angina (UA). Patients with ACS are at high risk of recurrent ischemic events and death. The incidence of 1-year composite adverse events (all-cause death / cardiovascular death, myocardial infarction, stroke and revascularization) is 11.3%, and the incidence of 4-year composite adverse events is 20.7%. Therefore, there is an urgent need for products to prevent and improve adverse cardiovascular events in ACS patients.

[0003] In addition, the current detection and characterization techniques of nitro fatty acids are relatively backward. Due to problems such as low endogenous concentration, poor stability and different distributions in tissues and biological fluids of nitro fatty acids, it is difficult to accurately quantify them in biological samples.

[0004] In view of this, the present invention is specifically proposed. Summary of the Invention

[0005] The purpose of the present invention is to provide the application of nitro fatty acids in predicting, preventing and improving adverse cardiovascular events in patients with acute coronary syndrome (ACS), so as to reduce the risk of adverse cardiovascular events in ACS patients.

[0006] In the first aspect, the present invention provides the application of nitro fatty acids in the products for preventing and improving adverse cardiovascular events in ACS patients; the nitro fatty acids are 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid.

[0007] In the second aspect, the present invention provides the application of products for detecting the content of nitro fatty acids or nitro fatty acids as markers in the preparation of products for predicting the risk of adverse cardiovascular events in ACS patients; the nitro fatty acids are 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid.

[0008] In the present invention, the ACS patients refer to patients with acute cardiac ischemia syndrome caused by a sudden reduction in myocardial blood supply. The adverse cardiovascular events are events involving all-cause death, readmission due to non-fatal myocardial infarction / unstable angina / revascularization after ACS revascularization. The risk of adverse cardiovascular events refers to the increased risk of all-cause death, non-fatal myocardial infarction or ischemia-driven urgent revascularization after ACS revascularization.

[0009] The content of the nitro fatty acid is inversely proportional to the risk of adverse cardiovascular events after ACS revascularization. Specifically, for the ACS patients, when the concentration of 12-nitro conjugated linoleic acid in the body is greater than 0.196 ng / ml, or the concentration of 9-nitro conjugated linoleic acid is greater than 0.296 ng / ml, the risk of adverse cardiovascular events is lower, that is, the risk of non-fatal myocardial infarction / unstable angina / revascularization readmission and death is lower.

[0010] In a third aspect, the present invention provides a method for screening the nitro fatty acid, comprising the following steps:

[0011] (1) Obtain a biological sample from an ACS patient and perform pretreatment on the biological sample;

[0012] (2) Use a mass spectrometry-based detection technique to measure the pretreated biological sample and detect the content of each nitro fatty acid in the biological sample;

[0013] (3) Divide the ACS patients into an adverse event group and a non-adverse event group according to whether adverse cardiovascular events occur, and use the nitro fatty acid with a statistically significantly higher content level in the adverse event group than in the non-adverse event group as a candidate marker;

[0014] (4) Divide the ACS patients into a low level, a medium level, and a high level according to the tertiles of the candidate marker level, compare the incidence of adverse events among the patients at the three levels, and after multi-factor correction, the nitro fatty acid with a statistically significantly lower risk of the main endpoint event in the high level group than in the low level group is determined as a marker for preventing and improving the risk of adverse cardiovascular events in ACS patients.

[0015] In one embodiment, the nitro fatty acids for detection include: 9-nitrooleic acid, 10-nitrooleic acid, 10-nitro linoleic acid, 9-nitro conjugated linoleic acid, 12-nitro conjugated linoleic acid.

[0016] In one embodiment, the markers are 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid.

[0017] In one embodiment, the primary endpoint events include any one or more of all-cause death, non-fatal myocardial infarction, and unplanned revascularization, or any combination thereof.

[0018] Fourthly, the present invention provides a method for detecting the content of nitro fatty acids in a biological sample, comprising the following steps:

[0019] (1) Pretreat the biological sample;

[0020] (2) Detect nitro fatty acids in the pretreated biological sample using a mass spectrometry-based detection technique;

[0021] (3) Calculate the content of nitro fatty acids according to the standard curve equation.

[0022] In step (1), the pretreatment is carried out as follows: add aqueous acetic acid solution and internal standard solution to the serum sample, vortex and mix well, centrifuge at 3000 rpm for 30 s at 4°C; add ethyl acetate containing 0.02% BHT to the centrifuge tube, incubate at 1500 r / min for 20 min at 4°C; centrifuge at 10,000 rpm for 10 min at 4°C; take the supernatant and dry it; dissolve the dried sample in a mixed solvent of methanol (MeOH) and isopropanol (IPA), vortex and mix well for measurement.

[0023] The preparation of the internal standard solution: Take the stock solution of 10-nitrooleic acid-d17 (10-NO2-OA-d17), add an acetonitrile / isopropanol mixed solvent (volume ratio of acetonitrile to isopropanol is 1:1), and prepare a standard product with a concentration of 0.5 μg / mL as the internal standard solution.

[0024] The preparation of the ethanol solution containing BHT: Weigh 0.01 g of BHT powder with a balance, dissolve it in 50 mL of ethyl acetate, and mix well.

[0025] As one of the specific embodiments of the present invention, the mass spectrometry-based detection technique is the LC-MS / MS method; the conditions for detecting by the LC-MS / MS method are:

[0026] Chromatographic column: C18;

[0027] Mobile phase A: water / acetonitrile / formic acid (volume ratio 63 / 37 / 0.02);

[0028] Mobile phase B: acetonitrile / isopropanol (volume ratio 1:1);

[0029] Flow rate: 0.3 mL / min;

[0030] Ion source temperature: 300°C;

[0031] Elution gradient:

[0032] Table 1 Elution Gradient

[0033] Time (min) Mobile phase A Mobile phase B Flow rate (mL / min) 0 100.0 0.0 0.3 2.0 100.0 0.0 0.3 7.0 60.0 40.0 0.3 12.0 60.0 40.0 0.3 13.0 5.0 95.0 0.3 15.0 5.0 95.0 0.3 15.10 100 0.0 0.3 17.0 100 0.0 0.3 。

[0034] In step (3), the standard curve is established according to the following operations: Plot the actual concentration against the peak area ratio of the nitro-fatty acid external / internal standard to construct the standard curve corresponding to each nitro-fatty acid.

[0035] In step (3), the internal standard used for establishing the standard curve is selected from 10-NO2-OA-d17.

[0036] In step (3), the gradient concentration standard solution used for establishing the standard curve is: 0.02 ng / mL, 0.05 ng / mL, 0.1 ng / mL, 0.2 ng / mL, 0.5 ng / mL, 1 ng / mL, 2 ng / mL, 5 ng / mL, 10 ng / mL, 20 ng / mL, 50 ng / mL, 100 ng / mL.

[0037] In a fifth aspect, the present invention provides a kit for predicting the risk of adverse cardiovascular events in ACS patients, and the kit includes a product for detecting the content of nitro-fatty acids.

[0038] The nitro-fatty acids are 9-nitro-conjugated linoleic acid and 12-nitro-conjugated linoleic acid.

[0039] The product is a reagent for detecting the content of nitro-fatty acids in a biological sample from an ACS patient. Preferably, the reagent for detecting the content of nitro-fatty acids in a biological sample from an ACS patient is a reagent for determining the content of nitro-fatty acids by LC-MS / MS method.

[0040] Preferably, the prediction is jointly made based on the concentrations of both 12-nitro-conjugated linoleic acid and 9-nitro-conjugated linoleic acid, that is, when the concentrations of both are higher than the threshold, the risk of adverse cardiovascular events in ACS patients is predicted to be low. More specifically, compared with ACS patients with a concentration of 12-nitro-conjugated linoleic acid less than 0.069 ng / ml and a concentration of 9-nitro-conjugated linoleic acid less than 0.012 ng / ml in the biological sample, the risk of adverse cardiovascular events in ACS patients with a concentration of 12-nitro-conjugated linoleic acid greater than 0.196 ng / ml and / or a concentration of 9-nitro-conjugated linoleic acid greater than 0.296 ng / ml in the biological sample is lower.

[0041] The kit further includes a recording carrier.

[0042] The biological sample is a body fluid sample, including but not limited to blood samples, serum samples, plasma samples, or the blood lipid fraction or blood lipoprotein fraction obtained therefrom, as well as urine samples and serous cavity effusion samples.

[0043] In a sixth aspect, the present invention provides a product for preventing or improving adverse cardiovascular events in ACS patients, comprising 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid.

[0044] In the present invention, the product for preventing and improving adverse cardiovascular events in ACS patients is selected from any one of drugs, foods, and health foods.

[0045] Preferably, the drug includes preparations administered by parenteral routes, preparations administered by digestive routes, or preparations administered by other routes.

[0046] More preferably, the preparations administered by parenteral routes include sterile injection solutions or infusion solutions, etc. The preparations administered by digestive routes include but are not limited to tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, etc. The preparations administered by other routes include but are not limited to preparations administered by inhalation, topical (i.e., transdermal), and mucosal (intranasal, vaginal, etc.) routes, and the preparations include but are not limited to eye drops, intranasal sprays, inhalation aerosol sprays, etc.; ointments, pastes, creams, gels, transdermal patches, etc.

[0047] In one embodiment, the product for preventing and improving adverse cardiovascular events in ACS patients is a drug, and the drug comprises 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid, as well as a pharmaceutically acceptable carrier, or consists of 9-nitro conjugated linoleic acid, 12-nitro conjugated linoleic acid, and a pharmaceutically acceptable carrier.

[0048] In another embodiment, the product for preventing and improving adverse cardiovascular events in ACS patients is a food or a health food, and the food or health food comprises 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid, as well as a physiologically acceptable carrier, or consists of 9-nitro conjugated linoleic acid, 12-nitro conjugated linoleic acid, and a physiologically acceptable carrier.

[0049] In a seventh aspect, the present invention further provides a method for preventing or improving adverse cardiovascular events in a subject, comprising the following steps: administering a prophylactically or therapeutically effective amount of 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid or a product comprising 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid to a subject in need thereof to prevent or improve the adverse cardiovascular events in the subject, wherein the subject is an ACS patient.

[0050] Compared with the prior art, the beneficial effects achieved by the present invention are:

[0051] 1. The present invention firstly proposes a new use of nitro fatty acids in preventing or improving adverse cardiovascular events in ACS patients. As a biomarker, it can play a timely and effective preventive or improving role in the risk of adverse cardiovascular events in ACS patients, greatly reducing the risk of adverse cardiovascular events in ACS patients.

[0052] 2. The present invention also establishes a method for simultaneously detecting the contents of multiple nitro fatty acids, which can effectively solve the problem of difficult accurate quantification in biological samples due to the low endogenous concentration, poor stability, and different distributions in tissues and biological fluids of nitro fatty acids.

[0053] 3. The detection method provided by the present invention also has the advantages of being able to distinguish nitro conjugated linoleic acid and nitro oleic acid isomers in biological samples, achieving accurate detection of both, using less plasma for detection, having a simple plasma sample pretreatment method, and not requiring operations such as derivatization. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 Kaplan-Meier curve graph for grouping ACS patients by nitro conjugated linoleic acid level; in the figure, (a) represents 9-nitro conjugated linoleic acid, and (b) represents 12-nitro conjugated linoleic acid.

[0055] Figure 2 Forest plot for predicting events in ACS patients grouped by nitro conjugated linoleic acid level; in the figure, (a) represents 9-nitro conjugated linoleic acid, and (b) represents 12-nitro conjugated linoleic acid.

[0056] Figure 3 Chromatogram of 5 nitro fatty acids.

[0057] Figure 4 ROC curve; in the figure, (a) and (b) respectively represent the ROC curves of 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid in the development cohort; (c) and (d) respectively represent the ROC curves of 9-nitro conjugated linoleic acid and 12-nitro conjugated linoleic acid in the validation cohort. DETAILED DESCRIPTION OF THE INVENTION

[0058] The present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited to the following embodiments.

[0059] The experimental methods used in the following examples are all conventional methods unless otherwise specified.

[0060] The reagents, materials, instruments, etc. used in the following examples can all be obtained from commercial channels unless otherwise specified.

[0061] The detection method of nitro-fatty acids in biological samples (the samples are body fluid samples, including but not limited to blood samples, serum samples, plasma samples, or the blood lipid part or blood lipoprotein part obtained therefrom, as well as urine samples and serous cavity effusion samples) in the following embodiments is carried out according to the following method:

[0062] 1) Liquid-liquid extraction of LC-MS / MS sample spiking.

[0063] 2) Detection is carried out on an ultra-high pressure liquid chromatography tandem mass spectrometry system ACQUITY UPLC I Class Plus (Waters) in tandem with a SCIEX 7500 (AB) triple quadrupole mass spectrometer.

[0064] 3) Convert the mass and count of the detected peaks obtained by mass spectrometry into a list of corresponding nitro-fatty acid concentrations. Generate a calibration curve to determine the dynamic quantitative range of the detected nitro-fatty acids.

[0065] Apply a strict threshold to separate background noise from actual nitro-fatty acid peaks. Control each sample and accept it only when the acceptance criteria are met. Convert the mass and count of the detected peaks into a list of corresponding nitro-fatty acid names, and perform quantification using the calibration curve according to the ratio of nitro-fatty acids to the internal standard, and obtain their concentrations from the sample volume. The calibration curve is prepared by mixing known amounts of internal standards with various nitro-fatty acid standards.

[0066] The sample extraction operation steps are as follows:

[0067] 1. Standard dilution

[0068] 1) Preparation of nitro-fatty acid stock solution

[0069] Prepare 5 nitro-fatty acids, namely 9-nitrooleic acid, 10-nitrooleic acid, 10-nitro-linoleic acid, 9-nitro-conjugated linoleic acid, and 12-nitro-conjugated linoleic acid.

[0070] Take 5 nitro-fatty acid mother liquors, add LCMS ethanol to prepare 5 nitro-fatty acid stock solutions with a concentration of 1000 ng / ml, and store them at -20 °C;

[0071] 2) Preparation of internal standard solution and diluent

[0072] Preparation of internal standard solution: Take the 10-nitrooleic acid-d17 stock solution (10-NO2-OA-d17), add an acetonitrile / isopropanol mixed solvent (volume ratio of acetonitrile to isopropanol is 1:1) to prepare a standard product with a concentration of 0.5 μg / mL as the internal standard solution.

[0073] Preparation of internal standard dilution solution: Take the internal standard solution and add acetonitrile / isopropanol mixed solvent to prepare an internal standard dilution solution with a concentration of 100 ng / ml.

[0074] 3) Preparation of mixed standard gradient solution

[0075] Preparation of mixed external standard solution: Take 10 μL of each of the above 5 nitro-fatty acid stock solutions and add 50 μL of acetonitrile / isopropanol mixed solvent (volume ratio of acetonitrile to isopropanol is 1:1) to prepare a mixed external standard solution with a concentration of 100 ng / ml.

[0076] Preparation of mixed external standard dilution solution: Take the mixed external standard solution and dilute it with acetonitrile / isopropanol mixed solvent to prepare mixed external standard dilution solutions with corresponding concentrations of 0.02, 0.05, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100 ng / mL.

[0077] Preparation of mixed standard gradient solution: Add the above internal standard dilution solution to the above mixed external standard dilution solution to obtain a mixed standard gradient solution with an internal standard concentration of 4 ppb.

[0078] Note: 1) The pure standard gradient solution is stored for one week. The pure standard gradient solution stored for more than one week cannot be used and should be re-prepared.

[0079] 2. Pretreatment of nitro-fatty acid samples:

[0080] 1) Transfer 200 μL of plasma to a new 1.5 mL Eppendorf tube (operate on ice);

[0081] 2) Add 4 μL of internal standard solution and 5 μL of 20% acetic acid (volume ratio of acetic acid to water is 1:4);

[0082] 3) After vortex mixing for 30 s, centrifuge at 3000 rpm at 4°C for 30 s;

[0083] 4) Add 750 μL of ethyl acetate containing 0.02% BHT to each sample and vortex mix for 2 min;

[0084] Preparation of ethanol solution containing BHT: Weigh 0.01 g of BHT powder on a balance, dissolve it in 50 mL of ethyl acetate, and mix well.

[0085] 5) Incubate the sample at 1500 r / min at 4°C for 20 min;

[0086] 6) Centrifuge the sample at 10,000 rpm at 4°C for 10 min;

[0087] 7) Take 500 μL of the supernatant and transfer it to a new centrifuge tube, and dry it using a nitrogen evaporator;

[0088] 8) Reconstitute with 50 μL of a mixed solvent of LCMS methanol (MeOH) and isopropanol (IPA) (volume ratio of MeOH:IPA is 3:2), add to the sample vial, and wait for measurement.

[0089] 3. High Performance Liquid Chromatography - Tandem Mass Spectrometry Detection

[0090] Detection steps:

[0091] Load the sample extract (50 μL) onto a Kinetex 1.7 μm C18 100A 100 * 2.1 mm chromatographic column at a column temperature of 40 °C for detection; specific data is analyzed using SCIEX OS operating software. Construct a standard curve by plotting the actual concentration against the peak area ratio of nitro fatty acid external standard / internal standard. Calculate the content of the corresponding nitro fatty acid in the sample based on the standard curve and the peak area ratio of nitro fatty acid / internal standard in the sample.

[0092] Detection conditions:

[0093] Chromatographic column: C18;

[0094] Mobile phase A: water / acetonitrile / formic acid (volume ratio 63 / 37 / 0.02);

[0095] Mobile phase B: acetonitrile / isopropanol (volume ratio 1:1);

[0096] Flow rate 0.3 mL / min;

[0097] The gradient is as follows:

[0098] Table 2 Mobile phase gradient settings

[0099]

[0100]

[0101] Ion source temperature is set to 300 °C;

[0102] The standard curve equation for 9 - nitrooleic acid is: y = 3.98897e 5 x - 2.59912e 5 ;

[0103] The standard curve equation for 10 - nitrooleic acid is: y = 3.57381e 5 x - 2.05327e 5 ;

[0104] The standard curve equation for 10 - nitrolinoleic acid is: y = 2.88101e 5 x - 2.37523e 5 ;

[0105] The standard curve equation of 9-nitro conjugated linoleic acid is: y = 2.87591e 5 x - 2.86375e 5 ;

[0106] The standard curve equation of 12-nitro conjugated linoleic acid is: y = 2.93848e 5 x - 1.77360e 5 .

[0107] x is the concentration and y is the mass spectrometry response signal intensity.

[0108] The linear range of nitro fatty acids is 0.02 - 100 ng / mL, and the standard curve r 2 > 0.995.

[0109] Figure 3 is the chromatogram of nitro fatty acids.

[0110] In addition, when the chromatographic column is newly replaced, for the detection of nitro fatty acids, it is necessary to first flush with 100% mobile phase B for 30 min, and then flush with 100% mobile phase A for 30 min; when the samples are not detected for a long time, the column is sealed with 100% mobile phase B.

[0111] 4. Sample Detection

[0112] The within-batch and between-batch precisions of nitro fatty acids are detected by detecting multiple concentrations within the dynamic range (where the concentrations of 10-nitro linoleic acid are 1 ng / mL and 50 ng / mL, and the concentrations of the remaining nitro fatty acids are 0.05 ng / mL, 1 ng / mL, and 50 ng / mL), with 6 samples at each concentration point.

[0113] The results show that:

[0114] The within-batch precisions of 9-nitrooleic acid corresponding to the three concentrations are 17.54%, 5.88%, and 7.3% respectively, and the between-batch precisions corresponding to the three concentrations are 11.93%, 5.99%, and 1.77% respectively.

[0115] The within-batch precisions of 10-nitrooleic acid corresponding to the three concentrations are 16.81%, 1.74%, and 4.36% respectively, and the between-batch precisions corresponding to the three concentrations are 17.96%, 2.44%, and 3.93% respectively.

[0116] The within-batch precisions of 10-nitro linoleic acid corresponding to the two concentrations (1 ng / mL and 50 ng / mL respectively) are 4.66% and 3.58% respectively, and the between-batch precisions corresponding to the two concentrations are 6.73% and 4.1% respectively.

[0117] The within - batch precision of 12 - nitro - conjugated linoleic acid for three concentrations was 13.3%, 3.1% and 2.7% respectively, and the between - batch precision for three concentrations was 9.5%, 5.8%, 3.9%.

[0118] The within - batch precision of 9 - nitro - conjugated linoleic acid for three concentrations was 9.8%, 6.6% and 3.4% respectively, and the between - batch precision for three concentrations was 11.1%, 4.4%, 2.7% respectively.

[0119] Example 1

[0120] 720 ACS patients were followed up. After an average of 4 years, 36 died of all causes, and 38 were readmitted due to non - fatal myocardial infarction / unstable angina pectoris / revascularization.

[0121] The outcome events were all - cause death, readmission due to non - fatal myocardial infarction / unstable angina pectoris / revascularization.

[0122] The experiment was as follows:

[0123] (1) ACS patients were divided into an adverse cardiovascular event group and a non - adverse cardiovascular event group according to the occurrence of adverse cardiovascular events; among them, there were 74 cases in the adverse cardiovascular event group and 646 cases in the non - adverse cardiovascular event group;

[0124] (2) The method of high - performance liquid chromatography - tandem mass spectrometry was used to measure the contents of 5 nitro - fatty acids in the plasma of patients in the adverse cardiovascular event group and the non - adverse cardiovascular event group. The results showed that only 3 nitro - fatty acids could be detected, namely 10 - nitro - oleic acid, 9 - nitro - conjugated linoleic acid and 12 - nitro - conjugated linoleic acid, and the other 2 nitro - fatty acids were not detected; at the same time, the nitro - fatty acids with significantly higher content levels in the adverse event group than in the non - adverse event group were used as candidate markers; the candidate markers were 9 - nitro - conjugated linoleic acid and 12 - nitro - conjugated linoleic acid;

[0125] (3) All patients were divided into low level (T1), medium level (T2), and high level (T3) according to the tertiles of the levels of 9 - nitro - conjugated linoleic acid and 12 - nitro - conjugated linoleic acid, and the occurrence of adverse cardiovascular events among the three groups of T1 / T2 / T3 patients was compared; the results were as Figure 1As shown by the Kaplan-Meier curves, among them, in the 12-nitro-conjugated linoleic acid T3 group (i.e., when the concentration is greater than 0.196 ng / ml), compared with the 12-nitro-conjugated linoleic acid T1 group (i.e., when the concentration is less than 0.069 ng / ml), the adverse cardiovascular events were lower (Log rank, P<0.001), and the cumulative non-event rate was higher; in the 9-nitro-conjugated linoleic acid T3 group (i.e., when the concentration is greater than 0.296 ng / ml), compared with the 9-nitro-conjugated linoleic acid T1 group (i.e., when the concentration is less than 0.012 ng / ml), the adverse cardiovascular events were lower (Log rank, P<0.001), and the cumulative non-event rate was higher.

[0126] After multivariate adjustment, as Figure 2 shown, the risk of adverse events in the 9-nitro-conjugated linoleic acid T3 group was 0.266 times that of the 9-nitro-conjugated linoleic acid T1 group (adjusted HR 0.266, 95% CI: 0.088 - 0.803, P = 0.010), and the risk of adverse events in the 12-nitro-conjugated linoleic acid T3 group was 0.266 times that of the 12-nitro-conjugated linoleic acid T1 group (adjusted HR 0.266, 95% CI: 0.097 - 0.729, P = 0.010), indicating that high levels of 9-nitro-conjugated linoleic acid and 12-nitro-conjugated linoleic acid were significantly associated with a decrease in adverse cardiovascular events. It is suggested that these two nitro-fatty acids can be used as markers to predict the risk of adverse cardiovascular events in ACS patients, and these two nitro-fatty acids can also be used to prevent or improve the adverse cardiovascular events in ACS patients.

[0127] Example 2

[0128] A cohort of 720 ACS patients was assigned to a development cohort and a validation cohort according to the admission time. Among them, the development cohort consisted of 413 ACS patients (57.3%) admitted from January to June 2019, with an incidence of adverse cardiovascular events (10.1%), and the validation cohort consisted of 307 ACS patients (42.7%) admitted from July to December 2019, with an incidence of adverse cardiovascular events (10.2%).

[0129] The receiver operating characteristic (ROC) curve was used to evaluate the efficacy of nitro-conjugated linoleic acid levels in predicting adverse cardiovascular events in ACS patients. The results of the ROC curve analysis are shown in Table 3. Low levels of 9-nitro-conjugated linoleic acid and 12-nitro-conjugated linoleic acid had sensitivity and specificity in predicting specific adverse cardiovascular events, and had significant statistical significance (both P<0.05).

[0130] Table 3 Predictive value of low levels of nitro-conjugated linoleic acid for adverse cardiovascular events in ACS patients

[0131]

[0132] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made thereto based on the present invention, which will be obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of the present invention claimed.

Claims

1. Use of nitro fatty acids in the preparation of a product for predicting the risk of adverse cardiovascular events in ACS patients; The nitro fatty acids are 9-nitro conjugated linoleic acid and / or 12-nitro conjugated linoleic acid.

2. The application according to claim 1, wherein The ACS patients are patients after ACS revascularization of acute ischemic syndrome of the heart caused by a sudden reduction in myocardial blood supply; The risk of adverse cardiovascular events includes an increased risk of all-cause death, non-fatal myocardial infarction, or ischemia-driven urgent revascularization after ACS revascularization.

3. A kit for predicting the risk of adverse cardiovascular events in ACS patients, comprising a reagent for detecting the content of nitro fatty acids; The nitro fatty acids are 9-nitro conjugated linoleic acid and / or 12-nitro conjugated linoleic acid.

4. The kit according to claim 3, characterized in that, The reagent is a reagent for detecting nitro fatty acids in a biological sample from an ACS patient.

5. The kit according to claim 4, wherein The reagent is a reagent for determining the content of nitro fatty acids by LC-MS / MS method.

6. The kit according to any one of claims 3-5, characterized in that, The kit further comprises a recording carrier.

7. Use of the kit according to any one of claims 3-6 in the preparation of a product for predicting the risk of adverse cardiovascular events in ACS patients.

8. The application according to claim 7, wherein The content of the nitro fatty acids is inversely proportional to the risk of adverse cardiovascular events after ACS revascularization.

9. The application according to claim 7 or 8, characterized in that, When the concentration of 12-nitro conjugated linoleic acid in the biological sample is greater than 0.196 ng / ml and / or the concentration of 9-nitro conjugated linoleic acid is greater than 0.296 ng / ml, it is predicted that the ACS patient has a low risk of adverse cardiovascular events.

10. The application according to claim 7 or 8, characterized in that Compared with ACS patients with a concentration of 12-nitro conjugated linoleic acid less than 0.069 ng / ml and a concentration of 9-nitro conjugated linoleic acid less than 0.012 ng / ml in the biological sample, ACS patients with a concentration of 12-nitro conjugated linoleic acid greater than 0.196 ng / ml and / or a concentration of 9-nitro conjugated linoleic acid greater than 0.296 ng / ml in the biological sample have a lower risk of adverse cardiovascular events.

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