Application of lactic acid phenylalanine in preparation of product for distinguishing healthy people from hypertensive patients and improving blood pressure of hypertensive patients

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of lactic acid phenylalanine in preparation of products for distinguishing healthy people from hypertensive patients and improving blood pressure of the hypertensive patients. The invention discloses lactic acid phenylalanine, a product for detecting lactic acid phenylalanine or application of lactic acid phenylalanine as a marker in preparation of a product for distinguishing healthy populations from hypertension patients. The invention provides application of lactic acid phenylalanine in preparation of products for preventing, improving or treating hypertension. Meanwhile, the invention further provides a detection method of lactic acid phenylalanine, and the accuracy of biological sample detection is improved. The content of lactic acid phenylalanine in a biological sample is detected, lactic acid phenylalanine is supplemented, and hypertension can be prevented, improved or treated.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to application of lactate phenylalanine in preparation of products for distinguishing healthy people from hypertension patients and improving blood pressure of hypertension patients. BACKGROUND

[0002] Hypertension refers to that, without using antihypertensive drugs, the systolic pressure is continuously greater than or equal to 130 mmHg or the diastolic pressure is greater than or equal to 80 mmHg, which is associated with increased risk of cardiovascular disease events, including coronary heart disease, heart failure and stroke, and increased risk of death. Detailed understanding of the mechanism leading to hypertension is crucial for effective prevention or management of hypertension. Blood pressure-lowering drug intervention is widely used to reduce blood pressure through various mechanisms, including promoting vasodilation, inhibiting the renin-angiotensin-aldosterone pathway and increasing diuresis to reduce blood volume. Therefore, there is a need for new plasma biomarkers to assist in reducing blood pressure in hypertension patients.

[0003] Lactate phenylalanine (Lac-Phe, N-lactoyl-phenylalanine), also known as N-lactoyl phenylalanine, is a conjugated product of lactic acid and phenylalanine, and is the most abundant among N-lactoyl amino acids. Exercise-induced lactate phenylalanine can reduce food intake and obesity, but the relationship between lactate phenylalanine and the onset and development of hypertension is not yet clear.

[0004]

[0005] Metabolites from gut microbiota can play a role in blood pressure regulation-related mechanisms through vasodilation, renal sodium / potassium regulation, immune regulation and neurotransmitter regulation. For example, phenylacetyl glutamine is a gut microbiota-dependent metabolite associated with cardiovascular disease and drives platelet reactivity and thrombosis through adrenergic receptors. Phenylacetyl glutamine directly induces endothelial cell activation by inducing oxidative stress, leading to endothelial dysfunction, which is an early event in the development of several cardiovascular diseases, but there is no public information on whether phenylacetyl glutamine is involved in the mechanism of hypertension.

[0006] Choline, L-carnitine and betaine are metabolized by gut microbiota to produce trimethylamine, which is rapidly oxidized by flavin-containing monooxygenase-3 to form trimethylamine N-oxide in the liver. Gamma-butyrobetaine is produced during the metabolism of L-carnitine by intestinal microbes and plays a role in promoting atherosclerotic intermediates in the production of trimethylamine N-oxide. Increasing evidence suggests that trimethylamine N-oxide and its precursors are important factors in hypertension, but the significance of the levels of the above substances as specific metabolites in hypertension patients is not yet certain.

[0007] Applicants believe that analysis of phenylalanine lactate, phenylacetylglutamine and trimethylamine N-oxide related precursors can provide valuable information about the relationship between metabolites and hypertension. SUMMARY

[0008] The present application aims to provide the use of phenylalanine lactate in the preparation of products for distinguishing healthy people from hypertensive patients and for improving blood pressure in hypertensive patients. The technical problems to be solved are not limited to the technical subject described, and other technical subjects not mentioned herein can be clearly understood by those skilled in the art through the following description.

[0009] In a first aspect, the present application provides the use of phenylalanine lactate, a product for detecting phenylalanine lactate in a biological sample or phenylalanine lactate as a marker in the preparation of a product for distinguishing a healthy population from a hypertensive population.

[0010] In one embodiment, the hypertensive population is a hypertensive human population. Specifically, the hypertensive human population is a patient with essential hypertension. Preferably, the hypertensive human population is a population with a systolic blood pressure of ≥ 130 mm Hg and / or a diastolic blood pressure of ≥ 80 mm Hg without the use of antihypertensive drugs.

[0011] In another embodiment, the distinguishing of the healthy population from the hypertensive population is that the plasma phenylalanine lactate level of the healthy population is significantly higher than that of the hypertensive population.

[0012] In one embodiment, the biological sample is a body fluid sample including but not limited to a blood sample, a serum sample, a plasma sample or a blood lipid fraction or a blood lipoprotein fraction obtained therefrom, and a urine sample, a serous cavity effusion sample.

[0013] In another embodiment, the product for detecting phenylalanine lactate in a biological sample is a product for determining the content of phenylalanine lactate in a biological sample by a mass spectrometry-based assay.

[0014] In one embodiment, the product for distinguishing a healthy population from a hypertensive population includes but is not limited to a reagent, a kit.

[0015] In another embodiment, the product further comprises a carrier with the following judgment criteria: if the phenylalanine lactate in the biological sample is less than 5.827 ng / mL, then the individual from which the biological sample is derived is classified into the hypertensive population.

[0016] In a second aspect, the present application provides a method for screening a marker for distinguishing a healthy population from a hypertensive population in vitro, comprising the following steps: (1) obtaining a biological sample from a subject, and pretreating the biological sample; (2) determining the level of metabolite in the pretreated biological sample by mass spectrometry-based assay; (3) dividing the subjects into healthy population and hypertension population according to their blood pressure levels without using blood pressure lowering drugs, and taking the metabolite whose level in the healthy population is statistically significantly different from that in the hypertension population as a candidate marker; (4) analyzing the level of the candidate marker by receiver operating characteristic curve, and taking the metabolite whose area under the curve is greater than 0.7 and has significant statistical significance as a marker.

[0017] In one embodiment, the mass spectrometry-based assay is liquid chromatography tandem mass spectrometry (LC-MS / MS) method.

[0018] In another embodiment, the marker screened for distinguishing the healthy population and the hypertension population is phenylalanine lactate.

[0019] In a third aspect, the present application provides a method for detecting the content of metabolite in a biological sample, comprising the following steps: 1) pretreating the biological sample; 2) detecting the metabolite in the pretreated biological sample by mass spectrometry-based assay; 3) calculating the content of the metabolite according to the standard curve equation.

[0020] In one embodiment, in step 1), the pretreatment is performed according to the following operation: adding methanol and internal standard working solution to the biological sample, vortexing and mixing; centrifuging at 4℃, 14000 rpm for 10 min; taking the supernatant for detection.

[0021] In another embodiment, the mass spectrometry-based assay is liquid chromatography tandem mass spectrometry (LC-MS / MS) method; the detection conditions of the LC-MS / MS method are as follows: chromatographic column: HSS T3 (1.8 μm, 100×3.0 mm); mobile phase A: aqueous solution containing 0.005% formic acid and 0.05 mM ammonium formate; mobile phase B: methanol; flow rate: 0.4 mL / min; ion source temperature: 650℃; elution gradient: .

[0022] In one embodiment, the preparation of the internal standard working solution: take phenylacetyl glutamine-d5 powder, gamma-butyrobetaine-d9 powder, phenylalanine-d8 powder, betaine-d3 powder, respectively, add to methanol to prepare a standard solution with a concentration of 2 μg / mL; take trimethylamine N-oxide-d9 powder, choline-d4 powder, respectively, add to methanol to prepare a standard solution with a concentration of 20 μg / mL; take creatinine-d3 powder, add to pure water to prepare a standard solution with a concentration of 20 μg / mL; take L-carnitine-d3 powder, add to methanol to prepare a standard solution with a concentration of 100 μg / mL; take 10 μL of the above concentration solution of phenylacetyl glutamine-d5 solution, 10 μL of gamma-butyrobetaine-d9 solution, 10 μL of phenylalanine-d8 solution, 10 μL of betaine-d3 solution, 10 μL of trimethylamine N-oxide-d9 solution, 10 μL of choline-d4 solution, 10 μL of creatinine-d3 solution, and 20 μL of L-carnitine-d3 solution, add to 910 μL of 50% methanol aqueous solution to prepare a mixed solution of 1000 μL as an internal standard solution.

[0023] Specifically, the internal standard working solution of phenylacetyl glutamine-d5, phenylalanine-d8, gamma-butyrobetaine-d9 and betaine-d3 is 20 ng / mL; the concentration of trimethylamine N-oxide-d9, choline-d4 and creatinine-d3 is 200 ng / mL; the concentration of L-carnitine-d3 is 2000 ng / mL.

[0024] In another embodiment, in step 3), the establishment of the standard curve is performed according to the following operation: plot the peak area ratio of the external standard / internal standard of lactate phenylalanine and other metabolites against the actual concentration to construct the standard curve corresponding to each metabolite.

[0025] In one embodiment, in step 3), the internal standard used for the establishment of the standard curve is as follows: the internal standard of lactate phenylalanine, phenylacetyl glutamine, trimethylamine N-oxide, choline, creatinine, betaine, gamma-butyrobetaine and L-carnitine corresponds to phenylalanine-d8, phenylacetyl glutamine-d5, trimethylamine N-oxide-d9, choline-d4, creatinine-d3, betaine-d3, gamma-butyrobetaine-d9 and L-carnitine-d3, respectively.

[0026] In another embodiment, in step 3), the gradient concentration standard solution used for the establishment of the standard curve is: 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000, 20000 ng / mL.

[0027] In a fourth aspect, the present application provides use of phenylalanine in the preparation of a product for preventing, ameliorating or treating hypertension.

[0028] In one embodiment, the hypertension is essential hypertension. Specifically, In another embodiment, the product comprises phenylalanine in a unit dose of 50 mg / kg and a pharmaceutically acceptable carrier.

[0029] In one embodiment, the ameliorating or treating of hypertension is a significant reduction in systolic or / and diastolic blood pressure.

[0030] In another embodiment, the product is selected from any one of a drug, a food and a health food.

[0031] In one embodiment, the product comprises a formulation for administration via the digestive tract, a formulation for administration via a route other than the digestive tract or a formulation for administration via other routes. Specifically, the formulation for administration via the digestive tract includes, but is not limited to, tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups and the like. The formulation for administration via a route other than the digestive tract includes, but is not limited to, sterile injection solutions or infusion solutions and the like. The formulation for administration via other routes includes, but is not limited to, formulations for administration via inhalation, topical (i.e., transdermal) and / or mucosal (intranasal, vaginal and the like) routes; the formulation includes, but is not limited to, eye drops, intranasal sprays, inhalation aerosol sprays and the like; ointments, pastes, creams, gels, transdermal patches and the like.

[0032] In a fifth aspect, the present application provides a method for preventing, ameliorating or treating blood pressure in a subject, comprising the step of: administering to the subject an effective amount of phenylalanine or a product comprising phenylalanine to reduce blood pressure in the subject.

[0033] In one embodiment, the subject can be a mammal (e.g., a rodent, a primate or a human).

[0034] In another embodiment, the subject is a hypertensive patient; the subject is a hypertensive patient when the subject has a systolic blood pressure of ≥ 130 mm Hg or / and a diastolic blood pressure of ≥ 80 mm Hg without taking blood pressure lowering drugs.

[0035] In one embodiment, the effective amount is an amount sufficient to prevent, ameliorate or treat hypertension in the subject.

[0036] In another embodiment, the route of administration can include administration via a digestive route, administration via a non-digestive route, or administration via another route; the administration via a digestive route includes, but is not limited to, oral administration, gastrointestinal administration, etc.; the administration via a non-digestive route includes, but is not limited to, intravenous injection or infusion, intramuscular injection, local targeted injection, etc.; the administration via another route includes, but is not limited to, inhalation, local (i.e., transdermal) and / or mucosal (intranasal, vaginal, etc.) administration.

[0037] Compared with the prior art, the present application has the following beneficial effects: 1. The present application first proposes a new use of phenylalanine lactate in improving the blood pressure of hypertensive patients / hypertensive model mice, which can effectively improve the blood pressure of hypertensive patients / hypertensive model mice in a timely manner.

[0038] 2. The present application also develops and verifies a rapid and sensitive liquid chromatography-mass spectrometry analysis method for simultaneously detecting the contents of eight metabolites such as phenylalanine lactate, which can effectively solve the problems of large sample usage and long liquid gradient time in detecting metabolites such as phenylalanine lactate.

[0039] 3. The detection method provided by the present application also has the advantages of high sensitivity, short liquid gradient time, less plasma usage, simple plasma sample pretreatment method, etc. The method can simultaneously measure the concentrations of phenylalanine lactate, phenylacetyl glutamine, trimethylamine N-oxide, choline, creatinine, betaine, gamma-butyl betaine and L-carnitine in a sample by using only 5 μL of human plasma sample and only 4.2 minutes of chromatography running time.

[0040] 4. The present application also discloses that phenylalanine lactate can regulate the systolic and diastolic blood pressure of hypertensive mice induced by angiotensin II. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 The level distribution diagram of eight metabolites such as phenylalanine lactate in healthy and hypertensive populations.

[0042] Figure 2 The systolic and diastolic blood pressures of mice in the sham operation group (n=10), the angiotensin II group (n=8) and the angiotensin II+phenylalanine lactate group (n=8).

[0043] Figure 3 The chromatogram of eight metabolites such as phenylalanine lactate. DETAILED DESCRIPTION

[0044] The application will be further described in conjunction with the specific embodiments. The examples given are only to illustrate the application, and are not intended to limit the scope of the application. The examples provided below can serve as a guide for further improvement by those of ordinary skill in the art, and do not in any way constitute a limitation on the application.

[0045] The experimental methods in the following examples are all conventional methods, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained commercially, unless otherwise specified.

[0046] The detection method of the eight metabolites, including phenylalanine lactate, etc., in the biological samples (the samples are body fluid samples, including but not limited to blood samples, serum samples, plasma samples or blood lipid fractions or blood lipoprotein fractions obtained therefrom, and urine samples, serous cavity effusion samples) in the following examples is performed according to the following method: 1) Sample protein precipitation pretreatment extraction.

[0047] 2) Detection is performed on a system equipped with ultra-high pressure liquid chromatography tandem mass spectrometry LC-40 (Shimadzu) tandem Triple Quad 7500 (SCIEX) triple quadrupole mass spectrometer.

[0048] 3) The area of the detected peak obtained by mass spectrometry is converted into a list of corresponding phenylalanine lactate metabolite concentrations. A standard curve is generated to determine the dynamic quantitative range of the detected phenylalanine lactate metabolites.

[0049] A strict threshold is applied to separate the background noise from the actual phenylalanine lactate metabolite peaks. Each sample is controlled, and only when the acceptance criteria are met can it be accepted. The mass-to-charge ratio of the detected peak is converted into a list of corresponding phenylalanine lactate metabolite names, and according to the peak area ratio of phenylalanine lactate metabolites to internal standards, a standard curve is applied for quantification, and their concentrations are obtained from the sample volume. The standard curve is prepared by mixing known amounts of internal standards with various phenylalanine lactate metabolite standards.

[0050] The sample extraction operation steps are as follows: 1. Standard dilution 1) Preparation of phenylalanine lactate metabolite stock solution Prepare eight metabolites, namely phenylalanine lactate, phenylacetyl glutamine, trimethylamine N-oxide, choline, creatinine, betaine, gamma-butyl betaine and L-carnitine.

[0051] Take the above eight metabolite powders, among which phenylalanine lactate, phenylacetyl glutamine and gamma butyl betaine are diluted with methanol to a concentration of 100 μg / mL stock solution, trimethylamine N-oxide, choline, betaine and L-carnitine are diluted with methanol to a concentration of 200 μg / mL stock solution, creatinine is diluted with pure water to a concentration of 200 μg / mL stock solution, and stored at -20℃.

[0052] 2) Preparation of internal standard solution and diluent Preparation of internal standard solution: Take phenylacetyl glutamine-d5 powder, gamma butyl betaine-d9 powder, phenylalanine-d8 powder, betaine-d3 powder and add them to methanol to prepare standards with a concentration of 2 μg / mL; Take trimethylamine N-oxide-d9 powder, choline-d4 powder and add them to methanol to prepare standards with a concentration of 20 μg / mL; Take creatinine-d3 powder and add it to pure water to prepare a standard with a concentration of 20 μg / mL; Take L-carnitine-d3 powder and add it to methanol to prepare a standard with a concentration of 100 μg / mL.

[0053] Preparation of internal standard diluent: Take 10 μL of phenylacetyl glutamine-d5 solution, 10 μL of gamma butyl betaine-d9 solution, 10 μL of phenylalanine-d8 solution, 10 μL of betaine-d3 solution, 10 μL of trimethylamine N-oxide-d9 solution, 10 μL of choline-d4 solution, 10 μL of creatinine-d3 solution, and 20 μL of L-carnitine-d3 solution, and add them to 910 μL of 50% methanol aqueous solution to prepare a mixed solution of 1000 μL as the internal standard diluent.

[0054] 3) Preparation of mixed standard gradient solution Preparation of mixed external standard solution: Take 2 μL of phenylacetyl glutamine stock solution, 2 μL of gamma butyl betaine stock solution, 2 μL of phenylalanine lactate stock solution, 10 μL of betaine stock solution, 10 μL of trimethylamine N-oxide stock solution, 10 μL of choline stock solution, 10 μL of creatinine stock solution, and 10 μL of L-carnitine stock solution, and add them to 44 μL of 50% methanol aqueous solution to prepare a mixed external standard solution of 100 μL.

[0055] Preparation of mixed external standard diluent: Take the mixed external standard solution and dilute it with a 1:1 mixed solvent of methanol:water to prepare mixed external standard diluents with concentrations of 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000, 20000 ng / mL.

[0056] Note: Pure standard gradient solutions are stable for one week. Pure standard gradient solutions that have been stored for more than one week must be re-prepared. Figure 3

[0057] 2. Pretreatment of 8 metabolite samples including lactate and phenylalanine: 1) Transfer 5 μL of sample to a new 1.5 mL centrifuge tube and add 5 μL of internal standard solution; 2) Add 50 μL of methanol and vortex mix for 1 minute; 3) The sample was then centrifuged at 4°C and 14,000 rpm for 10 minutes; 4) After centrifugation, take the supernatant and add it to the sample vial for testing.

[0058] 3. High-performance liquid chromatography-tandem mass spectrometry detection Specific testing steps: The sample extract (40 μL) was loaded onto an HSS T3 column (1.8 μm, 100 × 3.0 mm) at a column temperature of 40 °C for detection. The specific data were analyzed using SCIEX OS software. A standard curve was constructed by plotting the peak area ratio of external standard to internal standard of metabolites such as lactate phenylalanine against the actual concentration. The content of the corresponding metabolites in the sample was calculated based on the standard curve and the peak area ratio of metabolites such as lactate phenylalanine to internal standard in the sample.

[0059] Testing conditions: Column: HSS T3 (1.8μm, 100×3.0mm); Mobile phase A: An aqueous solution containing 0.005% formic acid and 0.05 mM ammonium formate; Mobile phase B: Methanol; Flow rate: 0.4 mL / min; The gradient is shown below: Table 1. Mobile phase gradient settings .

[0060] The ion source temperature was set to 650℃.

[0061] The standard curve equation for lactic acid phenylalanine is: y = 0.27375x + 0.27786; the standard curve equation for phenylacetylglutamine is: y = 0.03667x + 0.00288; the standard curve equation for trimethylamine N-oxide is: y = 0.00124x + 0.00388; and the standard curve equation for choline is: y = 5.76687e -4 x+9.98777e -4 The standard curve equation for creatinine is: y = 2.87581e -4 x+1.16901e -4The standard curve equation for betaine is: y = 0.00508x + 0.07133; the standard curve equation for γ-butylbetaine is: y = 0.01927x + 8.10642e -4 The standard curve equation for L-carnitine is: y = 0.04970x + 0.14804, where x is the concentration and y is the intensity of the mass spectrometry response signal.

[0062] The linear ranges for lactic acid phenylalanine and γ-butyl betaine are 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, and 500 ng / mL; the linear range for phenylacetylglutamine is 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, and 2000 ng / mL; and the linear range for trimethylamine N-oxide is 1, 2, 5, 10, 20, 50, 100, 200, 500, and 1000 ng / mL. The linear ranges for choline, creatinine, and betaine were 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000, and 20000 ng / mL; the linear range for L-carnitine was 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10000, and 20000 ng / mL; the standard curve r 2 All are greater than 0.99.

[0063] like Figure 3 As shown, Figure 1 Chromatograms of eight metabolites, including lactic acid and phenylalanine.

[0064] When replacing the column with a new one, for the detection of eight metabolites such as lactic acid and phenylalanine, the column should first be rinsed with 100% mobile phase B for 30 minutes, and then rinsed with 100% mobile phase A for 30 minutes. If the sample is not to be tested for a long time, the column should be sealed with 100% mobile phase B.

[0065] 4. Sample testing The precision of eight metabolites, including lactate phenylalanine, was determined by detecting multiple concentrations within the limit of quantitation and dynamic range (the concentrations of lactate phenylalanine, phenylacetylglutamine, and γ-butyl betaine were 0.1, 0.5, 10, and 200 ng / mL; the concentrations of trimethylamine N-oxide, choline, creatinine, and betaine were 1, 5, 100, and 2000 ng / mL; and the concentrations of L-carnitine were 0.1, 0.5, 20, and 2000 ng / mL). Six samples were collected at each concentration point and tested over five consecutive days.

[0066] The results show: The precision of the four concentrations of lactate phenylalanine were 14.24%, 9.09%, 5.87%, and 3.25%, respectively.

[0067] The precision of the four concentrations of phenylacetylglutamine were 12.77%, 8.21%, 6.25%, and 5.20%, respectively.

[0068] The precision of the four concentrations of trimethylamine N-oxide were 12.62%, 7.16%, 6.67%, and 4.56%, respectively.

[0069] The precision of the four concentrations of choline were 12.60%, 7.07%, 5.16%, and 2.92%, respectively.

[0070] The precision of the four concentrations of creatinine were 11.91%, 5.10%, 4.16%, and 3.66%, respectively.

[0071] The precision of the four concentrations of betaine were 10.03%, 5.90%, 5.31%, and 4.69%, respectively.

[0072] The precision of the four concentrations of γ-butyl betaine were 12.70%, 7.03%, 3.68%, and 5.65%, respectively.

[0073] The precision of the four concentrations of L-carnitine were 12.12%, 7.28%, 3.09%, and 4.49%, respectively.

[0074] Example 1 The experiment was conducted on 222 age- and sex-matched healthy individuals and 222 hypertensive patients, as detailed below: (1) The content of eight metabolites in the plasma of healthy people and hypertensive patients was determined by high performance liquid chromatography-tandem mass spectrometry. Metabolites with statistically significant differences in content levels between the two groups were selected as candidate biomarkers.

[0075] like Figure 2 As shown, the metabolites with significant differences in plasma levels between healthy individuals and hypertensive patients are lactate phenylalanine, trimethylamine N-oxide, choline, creatinine, and L-carnitine.

[0076] (2) The efficacy of using receiver operating characteristic (ROC) curves to evaluate the ability of the five differential metabolites, including lactate and phenylalanine, to distinguish between healthy individuals and hypertensive patients.

[0077] The results are shown in Table 2. The area under the ROC curve for low lactate phenylalanine levels in determining hypertension was greater than 0.7 and was statistically significant (all P < 0.05).

[0078] Table 2. ROC curve parameters for differentially metabolized metabolites .

[0079] Example 2 Based on the above results, the present applicant explored the potential therapeutic effect of phenylalanine lactate on hypertensive model mice. To study the effect of phenylalanine lactate on hypertension, a mouse model was established by continuous infusion of angiotensin II for 14 days. After implanting the osmotic micro-pump, C57BL / 6J male mice were randomly divided into a sham operation group (saline infusion), an angiotensin II group (angiotensin II infusion of 1000 ng / kg / min), or an angiotensin II + phenylalanine lactate group (angiotensin II infusion of 1000 ng / kg / min and phenylalanine lactate 50 mg / kg daily intraperitoneal injection).

[0080] A significant increase in blood pressure was observed on day 2, confirming the successful induction of the model. Interestingly, compared with the group using angiotensin II alone, in the phenylalanine lactate treatment group, after the initial pressor response, the blood pressure was significantly reduced from day 4, indicating the inhibitory effect of phenylalanine lactate on hypertension (as shown in ​

[0081] The above has been described in detail. For those skilled in the art, without departing from the purpose and scope of the present application, and without unnecessary experiments, the present application can be implemented in a wider range under equivalent parameters, concentrations and conditions. Although the present application gives a special example, it should be understood that further improvements can be made to the present application. In summary, according to the principle of the present application, the present application intends to include any change, use or improvement of the present application, including changes made by conventional techniques known in the art, which deviates from the range disclosed in the present application.​

Claims

1. Use of phenylalanine lactate, a product for detecting phenylalanine lactate in a biological sample or phenylalanine lactate as a marker in the manufacture of a product for distinguishing a healthy population from a hypertensive population.

2. Use according to claim 1, characterized in that, The hypertensive population is a population of people with hypertension, which is a population of people with systolic blood pressure ≥ 130 mm Hg or / and diastolic blood pressure ≥ 80 mm Hg without using antihypertensive drugs.

3. Use according to claim 1, characterized in that, The healthy population is a population of people with significantly higher plasma phenylalanine lactate levels than the hypertensive population.

4. Use according to claim 1, characterized in that, The product for detecting the content of phenylalanine lactate is a product for determining the content of phenylalanine lactate by mass spectrometry-based determination.

5. Use according to claim 4, characterized in that, The product further comprises a carrier recording the following judgment criteria: if the phenylalanine lactate in the biological sample is less than 5.827 ng / mL, the individual from which the biological sample is derived is classified into the hypertensive population.

6. A method for screening a marker for distinguishing a healthy population from a hypertensive population in vitro, comprising the following steps: (1) obtaining a biological sample from a subject, and pretreating the biological sample; (2) determining the level of metabolites in the pretreated biological sample by mass spectrometry-based determination; (3) dividing the subject into a healthy population and a hypertensive population according to the blood pressure level without using antihypertensive drugs, and taking the metabolite whose content level is statistically significantly different between the healthy population and the hypertensive population as a candidate marker; (4) analyzing the level of the candidate marker by using a receiver operating characteristic curve, and taking the metabolite with an area under the curve greater than 0.7 and having significant statistical significance as a marker.

7. The method of claim 6, wherein, The marker for distinguishing a healthy population from a hypertensive population is phenylalanine lactate.

8. A method of detecting the content of a metabolite in a biological sample, characterized in that, Comprising the following steps: 1) pretreating the biological sample; 2) detecting the metabolites in the pretreated biological sample by mass spectrometry-based determination; 3) calculating the content of the metabolites according to the standard curve equation.

9. Use of phenylalanine lactate in the manufacture of a product for preventing, improving or treating hypertension.

10. Use according to claim 9, characterized in that, The hypertension is essential hypertension.

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