Application of water-soluble tomato concentrate in preparation of product for improving heart health

Abstract

The invention belongs to the technical field of biological medicine and food, and particularly relates to application of a water-soluble tomato concentrate (WSTC) in preparation of a product for improving heart health. Researches and tests find that the water-soluble tomato concentrate can increase myocardial hemodynamic, reduce venous extravasated blood area, expand blood vessels, relieve myocardial ischemia and reduce myocardial cell injury and apoptosis, and can inhibit release of myocardial enzyme caused by myocardial infarction in myocardial infarction; inflammation factors and immune cell infiltration are reduced, oxidation resistance is improved, the blood fat level is reduced, and atherosclerosis and coronary heart disease are improved. The effects of the water-soluble tomato concentrate provide a new development thought for improving heart health, especially for preventing and treating heart failure, myocardial infarction, atherosclerosis and coronary heart disease. Meanwhile, the water-soluble tomato concentrate disclosed by the invention is used as a food-grade raw material, is safe and reliable, and is convenient to popularize and use.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and food technology, and in particular, the invention relates to application of water-soluble tomato concentrate in preparing products for improving heart health. Background Art

[0002] As one of the most important organs in the human body, the health of the heart is closely related to the quality of life. A healthy heart includes normal heart rate, electrocardiogram, good blood pressure control, normal ejection fraction, normal heart structure and good myocardial function. Common heart-related diseases include coronary heart disease, hypertension, cardiomyopathy, myocardial infarction, arrhythmia, heart failure, etc.

[0003] Heart failure (HF) is a complex clinical syndrome caused by abnormal changes in cardiac structure and (or) function due to various reasons, resulting in impaired ventricular systolic and (or) diastolic function, mainly manifested as dyspnea, fatigue and fluid retention (pulmonary congestion, systemic congestion and peripheral edema). According to the left ventricular ejection fraction (LVEF) and the changes after treatment, it is divided into heart failure with reduced ejection fraction (HFrEF), heart failure with improved ejection fraction (HFimpEF), heart failure with mildly reduced ejection fraction (HFmrEF) and heart failure with preserved ejection fraction (HFpEF). According to the time and speed of heart failure, it is divided into chronic heart failure and acute heart failure. Most patients with acute heart failure have partial relief of symptoms after hospitalization and are transferred to chronic heart failure. Coronary heart disease, hypertension, diabetes and obesity are all predisposing factors for heart failure, but these factors do not necessarily lead to heart failure. At present, heart failure is considered to be a chronic, spontaneously progressive disease. Neuroendocrine system activation leading to myocardial remodeling is a key factor in the occurrence and development of heart failure; myocardial metabolism, inflammation, adipose factors and cytokines also play an important role in the occurrence and development of heart failure; endothelial cell damage leads to insufficient secretion of nitric oxide (NO), which in turn leads to damage to the NO-soluble guanylate cyclase-cyclic guanosine monophosphate cell signaling pathway, which also has an important impact on myocardial fibrosis and myocardial remodeling; at the same time, mitochondrial function damage is also one of the important factors in the occurrence and development of heart failure. There are many clinical drugs for the prevention and treatment of heart failure, such as diuretics, renin-angiotensin system inhibitors, ACEI, β-blockers, and digitalis drugs. There are also some foods and Chinese medicine ingredients that are used for the prevention and treatment of heart failure, such as astragalus, ginseng, salvia miltiorrhiza, scutellaria baicalensis, polygonatum odoratum, cassia twig, etc. CN201610170045.2 discloses the use of aconite, scutellaria baicalensis, apocynum venetum, corn, ganoderma lucidum, ginseng, etc. for relieving heart failure; CN202211565184.7 discloses a dietary supplement composition for improving cardiac function, comprising (1) β-nicotinamide mononucleotide or trisodium 1,6-diphosphate fructose, and (2) coenzyme Q10 and multivitamins, and the composition is claimed to have the efficacy of preventing and treating heart failure. CN202410606371.8 discloses a Chinese medicine composition for preventing and / or treating chronic heart failure and its application, the composition comprising ginseng, astragalus, mulberry bark, scutellaria baicalensis, tuckahoe, umbellate, and ligusticum chuanxiong.

[0004] Myocardial infarction (abbreviated as "MI") is a disease caused by myocardial necrosis caused by persistent ischemia and hypoxia of the coronary arteries of the human heart. The main causes of myocardial infarction include atherosclerosis, insufficient myocardial oxygen supply, increased myocardial oxygen consumption, embolism, etc.; myocardial infarction is easily induced by overeating, emotional excitement, shock or dehydration, and constipation when straining to defecate. CN202310661055.6 discloses the use of a prescription of ginkgo leaves, red peony root, tomato, Panax notoginseng, Gynostemma pentaphyllum, Polygala tenuifolia, Acorus calamus, Astragalus, Gastrodia elata, Eucommia ulmoides, and Rhizoma Anemarrhenae to treat cardiovascular diseases. It has a therapeutic effect on cardiac ischemia / reperfusion injury in rats, can reduce the area of ​​myocardial infarction in rats with acute ischemic coronary heart disease, and reduce LDH and CK content. Tomato has the effects of promoting body fluid and quenching thirst; strengthening the stomach and digestion, lowering blood lipids and blood pressure, and anti-inflammatory, and is used for thirst and loss of appetite. The tomatoes used are fresh fruits of tomatoes, which are combined with ginkgo leaves and red peony roots to work.

[0005] Atherosclerosis is one of the main factors causing stroke, coronary heart disease and myocardial infarction. Atherosclerosis is a disease in which lipid plaques accumulate on the walls of medium and large arteries, leading to reduced or blocked blood flow. It is often associated with endothelial cell inflammatory damage, increased vascular permeability, cholesterol deposition in the blood on the vessel walls, infiltration of immune cells, especially macrophages, leukocyte activation to produce a large number of oxygen free radicals, LDL oxidation and other factors. In atherosclerosis, atherosclerotic plaques on the arterial walls may rupture and bleed, causing thrombosis. Once the thrombus completely blocks the coronary artery, it will cause severe myocardial ischemia and hypoxia, leading to myocardial infarction.

[0006] However, given the diversity of factors that affect heart health and the complexity of conditions such as heart failure, myocardial infarction, atherosclerosis, and coronary heart disease, further in-depth research is still needed on products that improve heart health and to provide prevention and treatment plans.

[0007] Water-soluble tomato concentrate (WSTC) is an extract from tomatoes developed by Provis Natural Products Ltd. in the UK. It is known as "natural aspirin" because of its ability to inhibit platelet aggregation. In 2009, EFSA (European Food Safety Authority) first approved a health claim study on "water-soluble tomato concentrate helps maintain normal platelet aggregation and contributes to healthy blood flow", and this health claim has been approved. Multiple test results have confirmed that water-soluble tomato concentrate contains naturally occurring antiplatelet compounds, which can inhibit platelet activity in healthy people and reduce platelet aggregation after ingestion, thereby keeping the blood in a fluid and hypocoagulable state, preventing the formation of microaggregates in the circulatory system and preventing platelets from adhering to blood vessel walls or fatty plaques, thereby maintaining healthy blood circulation. At the same time, platelet function is not completely inhibited and is maintained at an appropriate level, allowing platelets to aggregate when blood vessels are damaged, but will not disrupt the coagulation process necessary after injury. Currently, water-soluble tomato concentrate has been used in Europe as a nutritional supplement in the form of vitamins and minerals. The efficacy of water-soluble tomato concentrate in improving heart health has not been reported. Summary of the invention

[0008] Based on the deficiencies of the prior art, the inventors, through extensive research and screening, discovered that water-soluble tomato concentrate (WSTC) has the effect of improving heart health, especially in the prevention and treatment of heart failure and myocardial infarction, thereby completing the present invention.

[0009] In one aspect of the present invention, the present invention provides the use of a water-soluble tomato concentrate in a product for improving heart health. It can be understood by those skilled in the art that, as a product for improving heart health, it can play a role in regulating human functions, reducing the risk of disease occurrence, and thus play a role in the prevention, alleviation and / or treatment of diseases. On the one hand, the water-soluble tomato concentrate of the present invention can reduce the risk of heart-related diseases to a certain extent by regulating human heart functions in the presence of heart disease-inducing factors; at the same time, it can also inhibit the further progression of heart-related diseases, such as inhibiting the development of heart failure, myocardial infarction, atherosclerotic coronary heart disease and other diseases. On the other hand, the water-soluble tomato concentrate of the present invention can alleviate various symptoms caused by heart-related diseases. For example, it can alleviate the symptoms caused by heart failure, myocardial infarction, and coronary heart disease, including myocardial ischemia, chest pain, fatigue, palpitations, dyspnea, chest tightness, dizziness, etc.

[0010] In one embodiment, the present invention provides the use of a water-soluble tomato concentrate in a product for improving heart health, wherein the improvement of heart health is regulating the myocardial and vascular functions of the human heart; preferably, the regulation of the myocardial function of the human heart is increasing myocardial hemodynamics, inhibiting myocardial cell damage or apoptosis, reducing myocardial inflammatory factor infiltration, and enhancing myocardial antioxidant capacity.

[0011] In one embodiment, the present invention provides the use of a water-soluble tomato concentrate in a product for improving heart health, wherein improving heart health includes preventing, alleviating and / or treating heart-related diseases; in the present invention, heart-related diseases include increased incidence of stroke caused by imbalanced heart function, hypertension, heart failure, myocardial infarction, and atherosclerotic coronary heart disease.

[0012] Preferably, the heart-related disease is one or more of heart failure, myocardial infarction, and atherosclerotic coronary heart disease.

[0013] In one embodiment, the water-soluble tomato concentrate of the present invention is used to reduce the risk of heart failure, myocardial infarction, atherosclerosis, and coronary heart disease, prevent the occurrence of heart failure, myocardial infarction, and atherosclerotic coronary heart disease, and inhibit the progression and deterioration of unhealthy conditions; it has an alleviating and / or therapeutic effect on symptoms caused by heart failure, myocardial infarction, and atherosclerotic coronary heart disease.

[0014] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention improves myocardial hemodynamics, relieves myocardial ischemia, reduces myocardial cell damage and apoptosis, thereby inhibiting the progression to heart failure or aggravating the progression of heart failure.

[0015] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention improves the congestion symptoms in heart failure, namely improves the problems related to pulmonary congestion, systemic circulation congestion and peripheral edema.

[0016] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention is used to dilate blood vessels to prevent or treat heart failure.

[0017] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention inhibits the infiltration of inflammatory factors in the myocardium.

[0018] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention can significantly enhance the activity of antioxidant enzymes and reduce the factors causing heart disease due to excessive oxidative stress. Preferably, the activity of SOD and GSH-Px is increased and the level of MDA is reduced.

[0019] In one aspect of the present invention, the water-soluble tomato concentrate of the present invention inhibits myocardial collagen fiber deposition caused by myocardial infarction and reduces myocardial cell apoptosis.

[0020] In some embodiments of the present invention, the water-soluble tomato concentrate is prepared by crushing tomatoes, separating the water-soluble components therein by a physical method, and then concentrating and desugaring. Preferably, the physical method is to break up the tomatoes by a cold crushing method at a temperature not exceeding 60° C., and then separate the water-soluble fraction therein by centrifugation / filtration.

[0021] In some embodiments of the present invention, the water-soluble tomato concentrate mainly comprises nucleoside compounds, flavonoid compounds and phenolic acid compounds. Preferably, the total content of adenosine, chlorogenic acid and flavonoids in the water-soluble tomato concentrate is greater than or equal to 16 mg / g.

[0022] In some embodiments of the present invention, the product for improving heart health is in the form of food, health food, or medicine. The dosage forms of the product include: hard capsules, soft capsules, tablets, powders, gels, lozenges, sustained-release preparations, granule preparations, drops, pills, and liquid preparations.

[0023] At present, water-soluble tomato concentrates are managed as common foods in Europe, the United States, and China. Water-soluble tomato concentrates (WSTC) are mainly composed of heat-stable water-soluble compounds derived from tomatoes, which are their main biologically active ingredients. The main biologically active ingredients in the water-soluble tomato concentrate may come from the pulp of tomato fruits and / or the juice around tomato fruit seeds, and then the solid components, fat-soluble components and water-soluble sugars from the above sources are further removed, and then concentrated and dried to obtain the water-soluble tomato concentrate. That is, the water-soluble components are mainly separated from tomatoes by physical methods (centrifugation, filtration, etc.), and then concentrated and desugared to obtain the water-soluble components. Therefore, it can be understood that lycopene is a fat-soluble component, and therefore, the water-soluble tomato concentrate of the present invention does not contain or substantially does not contain lycopene.

[0024] The water-soluble compounds in the water-soluble tomato concentrate (WSTC) mainly include nucleoside compounds, flavonoid compounds and phenolic acid compounds as characteristic components. The nucleoside compound can be the nucleoside compound itself or the nucleotide formed by phosphorylation thereof, and the nucleoside compound may be selected from at least one of uridine, cytidine, adenosine and guanosine; the nucleoside compound is further preferably adenosine, adenosine-3'-monophosphate and / or adenosine-5'-monophosphate. The flavonoid compound can be the flavonoid compound itself or its glycosylation product, and the flavonoid compound may be selected from at least one of myricetin, luteolin, kaempferol and naringenin; the flavonoid compound is further preferably quercetin-3-O-glucoside, naringin and / or rutin. The phenolic acid compound can be a phenolic acid compound itself or a glycosylation product thereof, wherein the phenolic acid compound can be cinnamic acid or a derivative thereof, such as p-coumaric acid, caffeic acid, coumaric acid, ferulic acid, sinapinic acid; wherein the phenolic acid compound can be benzoic acid or a derivative thereof, such as p-hydroxybenzoic acid, chlorogenic acid, gallic acid, vanillic acid, syringic acid. It is understood that the bioactive components in WSTC are not limited to one or more of the above-mentioned types of substances, and may also contain other substances.

[0025] In addition, the water-soluble tomato concentrate can also be formulated with food or pharmaceutically acceptable excipients to improve its commercial attributes, including improved delivery, shelf life and taste, etc. These excipients include various sugars (monosaccharides, disaccharides, polysaccharides), flavoring agents, coloring agents, preservatives, thickeners, buffers, excipients, disintegrants, binders, lubricants.

[0026] It should be understood by those skilled in the art that the water-soluble tomato concentrate of the present invention can be purchased from Provis Natural Products Co., Ltd. or its authorized sales agency. In addition, patents WO 99 / 55350, WO 2010 / 049707 and CN102271697 also disclose methods for preparing water-soluble tomato concentrates that can be used in the present invention. Therefore, the water-soluble tomato concentrate used in the present invention can also be prepared by itself according to WO 99 / 55350, WO 2010 / 049707 or CN102271697. If prepared by itself according to the methods described in WO 99 / 55350, WO 2010 / 049707 or CN102271697, it should be ensured that the ingredients of the prepared water-soluble tomato concentrate meet the above requirements. In one embodiment, according to the method of CN102271697 Figure 2 and Figure 4 The process described is a preferred method for producing a water-soluble tomato concentrate.

[0027] In one embodiment, the water-soluble tomato concentrate used in the present invention can be prepared by the method described in CN102271697, the method comprising:

[0028] (a) preparing an initial mixture of tomato homogenate;

[0029] (b) separating the water-soluble component from the initial mixture, the separation temperature being no higher than 60° C.;

[0030] (c) filtering and concentrating the water-soluble components, wherein the concentration temperature is not higher than 60°C.

[0031] Preferably, the temperature of the initial mixture in step (a) is not higher than 35°C.

[0032] Preferably, the pH of the initial mixture in step (a) does not exceed 5.5.

[0033] Preferably, the browning index of the initial mixture does not exceed 0.4 AU at 4% solids, wherein the browning index is defined as the absorbance of the soluble fraction at 420 nm.

[0034] Preferably, in step (b), water-insoluble solids with a particle size greater than 500 μm are separated and removed from mixture A to obtain a water-soluble component.

[0035] Preferably, the separation in step (b) comprises at least one of centrifugal separation and natural sedimentation separation, and the water-soluble component is the supernatant obtained after separation.

[0036] Preferably, in step (c), the filtration is performed using a filter membrane. Without limitation, the pore size of the filter membrane used for filtration is not greater than 1 μm, specifically 0.2 μm.

[0037] Preferably, the concentration in step (c) can be carried out by membrane filtration concentration, low temperature evaporation concentration, freeze drying concentration or spray drying concentration.

[0038] Preferably, in the step (c), the step of removing free sugars from the water-soluble components before filtering is also included.

[0039] In one embodiment, the present invention discloses the use of a water-soluble tomato concentrate in the preparation of a product for improving heart health, wherein the product also includes other substances that have the effects of preventing, alleviating, improving, and treating adverse heart conditions.

[0040] In one embodiment, the other substances that have preventive, alleviating, improving and therapeutic effects on adverse cardiac conditions include coenzyme Q10, PQQ, NMN, NAD+, turmeric extract, astragalus extract, mulberry bark, scutellaria baicalensis extract, Poria extract, ginkgo leaf extract, ginseng extract, American ginseng extract, rhodiola rosea extract, squalene, ganoderma lucidum extract, vitamins, minerals, grape seed extract, bilberry extract, lotus leaf extract, lotus seed extract, wolfberry extract, red date extract, aspirin, red yeast extract, metformin, ergothioneine, one or more. Preferably, the substance is aspirin; more preferably, the substance is ginkgo leaf extract and ginseng extract.

[0041] In one embodiment, the water-soluble tomato concentrate and aspirin are used to prepare a composition for preventing and treating myocardial infarction.

[0042] In one embodiment, water-soluble tomato concentrate, ginkgo leaf extract and ginseng extract are used to prepare a composition for preventing and treating atherosclerotic coronary heart disease.

[0043] In one embodiment, the water-soluble tomato concentrate of the present invention may be used in combination with other agents that improve heart health.

[0044] In one embodiment, the combined use is to administer the water-soluble tomato concentrate of the present invention to a high-risk group of heart failure / myocardial infarction while taking a drug for preventing heart failure or myocardial infarction; or to administer the water-soluble tomato concentrate of the present invention to a subject with acute or chronic heart failure or myocardial infarction while taking a drug for treating heart failure or myocardial infarction. The administration method is preferably oral administration, which can be taken at the same time or at different times of the same day.

[0045] Beneficial Effects

[0046] The present invention unexpectedly found that the water-soluble tomato concentrate is used to improve heart-related diseases, especially heart failure, myocardial infarction, and atherosclerotic coronary heart disease. The water-soluble tomato concentrate of the present invention can increase the blood flow velocity and cardiac output of the heart, reduce the area of ​​venous congestion, and at the same time play the role of dilating blood vessels, which is beneficial to the prevention and treatment of heart failure; it can increase the activity of antioxidant enzymes, reduce myocardial cell apoptosis and myocardial fibrosis, reduce inflammatory factor infiltration and anti-oxidation, lower blood lipids, reduce the risk of myocardial infarction and coronary atherosclerosis, and reduce the adverse effects caused by the occurrence of myocardial infarction and coronary heart disease.

[0047] The water-soluble tomato concentrate of the present invention is used in combination with aspirin, and the two have a synergistic effect, improve heart / blood flow health, and reduce the side effects of medication. The water-soluble tomato concentrate of the present invention can be used in combination with ginkgo leaf extract and ginseng extract to play a synergistic role, enhance antioxidant capacity, reduce serum lipids and inflammation levels, inhibit lipid deposition and plaque formation of cholesterol, and have the effect of preventing and treating coronary atherosclerosis and coronary heart disease.

[0048] The water-soluble tomato concentrate of the present invention is a food-grade raw material, does not produce acute, subacute or chronic toxicity to the human body, is safe and reliable, provides a new idea for the prevention and treatment of heart health-related diseases such as heart failure, myocardial infarction, atherosclerosis, etc., and is easy to promote and use. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 : The blood flow velocity test results of zebrafish after different treatments, among which, compared with the model control group, **p<0.01, ***p<0.001;

[0050] Figure 2 : Test results of zebrafish cardiac output after different treatments, among which, compared with the model control group, *p<0.05, **p<0.01, ***p<0.001;

[0051] Figure 3: Shows typical graphs of zebrafish heart area and venous congestion area after treatment in different groups, where the red dotted box represents the zebrafish heart area, and the yellow dotted box represents the zebrafish venous congestion area.

[0052] Figure 4 : The results of zebrafish heart area test after different groups of treatment, among which, compared with the model control group, ***p<0.001;

[0053] Figure 5 : The test results of zebrafish venous congestion area after different groups of treatment, among which, compared with the model control group, **p<0.01, ***p<0.001;

[0054] Figure 6 : Typical diagram of zebrafish blood vessel diameter after treatment in different groups;

[0055] Figure 7 : The test results of zebrafish vascular diameter after different treatments, among which, compared with the model control group, *p<0.05, ***p<0.001.

[0056] Figure 8 : To detect the effects of WSTC on the appearance, weight and organ index of the rat heart; A is the measurement of the heart size of each group; B is the comparison of the heart weight of each group; C is the comparison of the heart organ index.

[0057] Fig. 9 : The effects of WSTC on various serum myocardial enzymes were detected, where A was LDH, B was CK, C was CK-MB, D was cTnI, and E was cTnT. Compared with the control group (control), #p<0.05, ##p<0.01, ###p<0.001; compared with the model group (ISOModel), *p<0.05, **p<0.01, ***p<0.001; compared with the combination group (WSTC+ASP), $p<0.05, $$$p<0.001.

[0058] Fig.10 :To detect the effect of WSTC on inflammatory factors, where A is TNF-α, B is IL-1β, and C is IL-6; compared with the control group (control), #p<0.05, ###p<0.001; compared with the model group (ISO Model), *

[0059] p<0.05, **p<0.01, ***p<0.001.

[0060] Fig.11: Detect the effect of WSTC on antioxidant enzyme activities and MDA levels, where A is SOD, B is GSH-Px, C is CAT, and D is MDA; compared with the control group (control), ###p<0.001; compared with the model group (ISO Model), *p<0.05, **p<0.01, ***p<0.001.

[0061] Fig.12 :Effects of WSTC on cardiac tissue pathology (HE staining).

[0062] Fig.13 : Effects of WSTC on cardiac tissue pathology (MASSON staining), compared with the control group (control), ###p<0.001; compared with the model group (ISO Model), ***p<0.001.

[0063] Fig.14 :The effect of water-soluble tomato concentrate (WSTC) on cardiomyocyte apoptosis, where A is the TUNEL staining of cardiomyocytes in each group, and B is the statistical analysis of apoptotic cells in each group; compared with the control group (control),

[0064] ###p<0.001; compared with the model group (ISO Model), *p<0.05, **p<0.01, ***p<0.001.

[0065] Fig.15 : Masson staining of mouse aorta paraffin sections, where A is the model control group; B is the positive drug group; C is sample group 1; D is sample group 2; E is sample group 3. DETAILED DESCRIPTION

[0066] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0067] The water-soluble tomato concentrate used in the examples of this application was purchased from Provis Natural Products Co., Ltd. Make sales.

[0068] Embodiment 1:

[0069] 1. Sample

[0070] (1) Sample ①: Water-soluble tomato concentrate The powder is diluted with purified water.

[0071] (2) Positive control: digoxin, white powder, batch number X5NPO-TP, TCI (Shanghai) Chemical Industry Development Co., Ltd., solvent DMSO.

[0072] (3) Adenosine, white powder, batch number WXBD7718V, Sigma, solvent is standard dilution water.

[0073] 2. Animals

[0074] Zebrafish were raised in fish farming water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), bred and provided by Huante Biological Fish Farming Center, the experimental animal use license number is: SYXK (Zhejiang) 2022-0004, and the breeding management complies with the requirements of the international AAALAC certification (certification number: 001458), IACUC ethics review number: IACUC-2024-8434-01.

[0075] 3. Related tests

[0076] 3.1 Evaluation of heart failure improvement efficacy (hemodynamics)

[0077] 2dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples were given water-soluble, and the positive controls were 0.8μg / mL digoxin and 20.0μg / mL adenosine. A normal control group and a model control group were also set up, with a capacity of 3mL per well. After 4h of sample treatment, except for the normal control group, all other experimental groups were given water-soluble verapamil hydrochloride to establish a zebrafish heart failure model. After treatment at 28℃ for 40min, 10 zebrafish were randomly selected from each experimental group and placed under a heartbeat blood flow analyzer to record videos. ViewPoint Application Manager software was used to collect data, and the blood flow velocity and cardiac output of zebrafish were analyzed. The statistical analysis results of the above indicators were used to evaluate the heart failure improvement efficacy of the samples (the results are shown in Tables 1 and 2). Figure 1-2 The statistical results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0078] Table 1 Results of the evaluation experiment on the efficacy (hemodynamics) of improving heart failure of samples (n=10)

[0079]

[0080] Compared with the model control group, * indicates p<0.05, ** indicates p<0.01, *** indicates p<0.001; compared with the digoxin group, # Indicates p<0.05; sample ① compared with the adenosine group, △ Indicates p<0.05.

[0081] The test results showed that compared with the normal control group, after the modeling of Verapamil hydrochloride, the blood flow velocity and cardiac output of the model group were significantly slowed down, with statistical differences, indicating that the heart failure model was successfully established. After the administration of digoxin, adenosine, and sample ①, the blood flow velocity and cardiac output were significantly increased, and there were statistical differences compared with the model group, indicating that the sample group can improve the symptoms of heart failure, specifically manifested in the improvement of blood flow velocity and cardiac output. Moreover, the high-dose group of sample ① significantly outperformed the adenosine group in promoting blood flow velocity, and the high-dose group of sample ① was significantly better than the digoxin group and adenosine group in improving cardiac output.

[0082] 3.2 Evaluation of efficacy in improving heart failure

[0083] 2dpf wild-type AB strain zebrafish were randomly selected in a 6-well plate, with 30 fish per well. The samples were given water-soluble (concentrations are shown in Table 6-3), and the positive control digoxin was 0.800μg / mL. At the same time, a normal control group and a model control group were set up, with a capacity of 3mL per well. After 4h of sample treatment, except for the normal control group, all other experimental groups were given 20.0μg / mL verapamil hydrochloride in water-soluble form to establish a zebrafish heart failure model. After 40min of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and placed under a dissecting microscope for photography. The data were analyzed and collected using NIS-Elements D3.20 software. The heart area and venous congestion area of ​​zebrafish were analyzed, and the statistical analysis results of the above indicators were used to evaluate the heart failure improvement efficacy of the samples (the results are shown in Table 2 below). The statistical analysis results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0084] Table 2 Results of the evaluation experiment on the efficacy of samples in improving heart failure (heart area and venous congestion area) (n=10)

[0085]

[0086] Compared with the model control group, * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001; compared with the digoxin group, # Indicates p<0.05; Sample ① compared with the adenosine group, △ Indicates p<0.05.

[0087] The test results showed that compared with the normal control group, after the modeling with Verapamil hydrochloride, the heart area and venous congestion area of ​​the model group increased significantly, with statistical differences, indicating that the heart failure model was successfully established. After the administration of digoxin, adenosine, and sample ①, the heart area and venous congestion area decreased significantly, with statistical differences compared with the model group, indicating that the sample group can improve the symptoms of heart failure, specifically in terms of the improvement of heart area and venous congestion. Among them, the effects of low and high doses of sample ① were significantly better than those of the adenosine group in terms of the improvement of heart area, and the high dose of sample ① was significantly better than those of the digoxin and adenosine groups in terms of the improvement of venous congestion area.

[0088] 3.3 Vascular dilation ability test

[0089] 2dpf red vascular fluorescent zebrafish were randomly selected in a 6-well plate, with 30 fish per well. The samples were given in water, and the positive control was 8.53μg / mL of calycosin (Huazhong Haiwei (Beijing) Gene Technology Co., Ltd.). At the same time, a normal control group was set up, with a capacity of 3mL per well. One day after the sample was treated, 10 zebrafish were randomly selected from each experimental group and placed under a Zeiss fluorescence microscope for photography. The data were collected using NIS-Elements D 3.20 software to analyze the diameter of the zebrafish intersegmental blood vessels. The statistical analysis results of the above indicators were used to evaluate the vasodilation effect of the samples (the results are shown in Table 3 below). The statistical processing results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant.

[0090] Table 3 Sample vasodilator efficacy evaluation test results (n = 10)

[0091]

[0092] Compared with the model control group, * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001; compared with the calycosin group, # Indicates p<0.05; Sample ① compared with the adenosine group, △△△ Indicates p<0.001.

[0093] The test results show that compared with the normal control group, the diameter of the intersegmental blood vessels of zebrafish increased significantly after administration of samples of calycosin isoflavones and adenosine, which was statistically different from the normal control group, indicating that sample group ① can dilate blood vessels with a significant effect, among which the high-dose group was better than the adenosine group.

[0094] In summary, the use of water-soluble lycopene in the present invention can improve the hemodynamics of heart failure, reduce the area of ​​venous congestion, dilate blood vessels, play a relevant role in the prevention and treatment of heart failure, inhibit the progression of the disease to heart failure or alleviate the symptoms of heart failure.

[0095] Example 2: Protective effect of water-soluble tomato concentrate (WSTC) on myocardial infarction

[0096] 2.1 Test animals

[0097] Healthy male SD rats (weight 180-200 g) were purchased from Beijing Weitonglihua Experimental Animal Company with license number SCXK (Beijing) 2021-0006 and certificate number: NO.110011241104862667.

[0098] 2.2 Animal grouping, drug treatment, and model establishment

[0099] The experimental animals were divided into: (1) sham operation group (control); (2) acute myocardial infarction (ISO) model group; (3) WSTC 12.5 mg / kg group; (4) WSTC 25 mg / kg group; (5) WSTC 50 mg / kg group; (6) aspirin (ASP, 10 mg / kg, clinical equivalent dose) group; (7) WSTC + ASP (25 mg / kg + 10 mg / kg) combined administration group. There were 10 rats in each group. Each administration group was intragastrically administered 14 days before ISO model establishment, while the control group and ISO model group rats were intragastrically administered with an equal volume of solvent. One hour after administration on the 13th and 14th days, the model group and each administration group rats were intraperitoneally injected with 85 mg / kg ISO (isoproterenol hydrochloride), and the control group was given an equal volume of solvent.

[0100] Test equipment and instruments:

[0101] Isoproterenol hydrochloride (ISO) (Cat. No.: I5627-5G): Sigma-Aldrich (Shanghai) Trading Co., Ltd. Aspirin enteric-coated tablets (Batch No. BJ51822): Bayer HealthCare Co., Ltd. Biosino Biopharmaceuticals Co., Ltd.: CK-MB kit (Batch No.: 241221), CK kit (Batch No.: 241421), LDH kit (Batch No.: 241011). Beijing Huaying Biotechnology Research Institute: Rat IL-6 ELISA Kit (Batch No.: HY-H0007), Rat TNF-α ELISA Kit (Batch No.: HY-H0019), Rat IL-1β ELISA Kit (Batch No.: HY-H0001), Superoxide Dismutase (SOD, Catalog No.: HY-M0001), Glutathione-peroxidase (GSH-PX, Catalog No.: HY-M0004), Malondialdehyde (MDA, Catalog No.: HY-M0003), Catalase (CAT, Catalog No.: HY-0018), Cardiac Troponin I (cTnI, Catalog No.: HY-D0022), Cardiac Troponin T (cTnT, Catalog No.: HY-D0019). Electronic balance (Model: AL104-IC): Mettler-Toledo Instruments (Shanghai) Co., Ltd. Ultrapure water instrument (model: Milli-Q): Shenzhen Mindray Bio-Medical Electronics Co., Ltd.

[0102] 2.3 Effects of water-soluble tomato concentrate (WSTC) on the appearance of rat hearts

[0103] See also Figure 8 In the experiment, the hearts of rats in the control group were smooth, round and uniform in color; the hearts of rats in the ISO model group had scattered gray-white patches concentrated at the apex of the heart, indicating that the model was successful; compared with the ISO model group, the appearance of the hearts of rats in the drug-treated group was improved.

[0104] 2.4 Effect of water-soluble tomato concentrate (WSTC) on serum myocardial enzymes

[0105] LDH, CK, and CK-MB are commonly used clinical biomarkers of myocardial injury. In particular, CK-MB only exists in myocardial cells and is considered a specific marker of myocardial injury. It is one of the gold standards for the diagnosis of myocardial injury. Cardiac troponin (cTn) is a group of proteins that regulate myocardial muscle contraction. Among them, cTnT and cTnI are myocardial-specific antigens with good myocardial specificity. When myocardial cells are damaged, cell permeability increases, troponin is released from myocardial fibers, causing an increase in serum troponin. It has now become the preferred marker for diagnosing acute myocardial infarction (AMI).

[0106] Blood was collected from the abdominal aorta of each group of rats to detect serum myocardial enzymes LDH, CK, CK-MB, cTnI, and cTnT. Fig. 9 As shown in the data, compared with the control group, LDH, CK, CK-MB, cTnI and cTnT in the ISO model group were significantly increased (p<0.001, p<0.05, p<0.01, p<0.001 and p<0.001), indicating that ISO caused significant myocardial damage; compared with the ISO model group, WSTC 25mg / kg group and WSTC 50mg / kg group could significantly inhibit the increase of CK, CK-MB and cTnI (p<0.05 or p<0.001) and were dose-dependent; WSTC 50mg / kg group could significantly reduce the levels of LDH and cTnT (p<0.001); the above results showed that WSTC could significantly reduce the release of myocardial enzymes caused by myocardial injury, and the effect of 50mg / kg WSTC high-dose group on reducing the release of myocardial enzymes was comparable to that of aspirin, and even better than ASP in some indicators. It is particularly noteworthy that the CK-MB level in the WSTC+ASP combined administration group was significantly lower than that in the WSTC 12.5 mg / kg and ASP 10 mg / kg groups (p<0.001 and p<0.05); the CK level in the WSTC+ASP combined administration group was significantly lower than that in WSTC 12.5 mg / kg (p<0.001), indicating the synergistic effect between WSTC and aspirin.

[0107] 2.5 Effect of water-soluble tomato concentrate (WSTC) on inflammatory factors

[0108] Inflammatory cell infiltration appears in the early stage of acute myocardial injury, indicating that inflammatory response plays an important role in the occurrence and development of AMI. TNF-α, IL-1β and IL-6 are important regulatory factors in the inflammatory response process.

[0109] Blood was collected from the abdominal aorta of rats in each group to detect inflammatory factors TNF-a, IL-1β, and IL-6. Fig.10As shown: Compared with the control group, the levels of serum inflammatory factors TNF-α, IL-1β and IL-6 in the ISO model group were significantly increased (p<0.001); compared with the ISO model group, the levels of TNF-α and IL-1β in the WSTC 12.5mg / kg group, WSTC 25mg / kg group and WSTC 50mg / kg group were significantly decreased (p<0.001, p<0.05 or p<0.01), the level of IL-6 in the WSTC 50mg / kg group was significantly decreased (p<0.05), and the levels of TNF-α and IL-6 in the ASP group were significantly decreased (p<0.05, p<0.01). The results suggest that the protective effect of WSTC on acute myocardial injury is related to the inhibition of inflammatory response, which significantly reduces the levels of TNF-α and IL-1β, and can also reduce the level of IL-6 under high dose conditions.

[0110] 2.6 Effects of water-soluble tomato concentrate (WSTC) on antioxidant enzyme activities and MDA levels

[0111] See also Fig.11 As shown in the figure, compared with the control group, the activities of SOD, GSH-Px and CAT in the ISO model group were significantly decreased (p<0.001), and the level of MDA was significantly increased (p<0.001); compared with the ISO model group, the SOD activity in the WSTC 12.5mg / kg group, WSTC 25mg / kg group and WSTC 50mg / kg group was significantly increased (p<0.05 or p<0.01), the GSH-Px activity in the WSTC 25mg / kg group and WSTC 50mg / kg group was significantly increased (p<0.05 or p<0.01), and the level of MDA was significantly decreased (p<0.001 or p<0.01); there was no significant difference in CAT activity between the WSTC dose groups and the ISO model group. SOD, GSH-Px and CAT are important lines of defense for the body's antioxidant system, MDA is a stable metabolite of reactive oxygen, and the decrease in antioxidant enzyme activity and the increase in MDA levels are significant signs of oxidative stress. The results of the present invention suggest that WSTC plays an antioxidant role in protecting against acute myocardial injury, enhancing the activity of antioxidant enzymes and reducing oxidative stress.

[0112] 2.7 Effects of water-soluble tomato concentrate (WSTC) on cardiac tissue pathology

[0113] The hearts of mice in each group were taken, sliced ​​and analyzed by HE staining. Fig.12The myocardial tissue morphology and structure of rats in the control group were normal, with neat arrangement, clear texture, uniform cell size and clear boundaries; the myocardial tissue structure of the ISO model group was loose and disordered, with enlarged gaps, a large number of inflammatory cells infiltrating, uneven cell size and unclear boundaries; compared with the model group, the myocardial tissue structure of the WSTC25mg / kg group and the WSTC50mg / kg group was significantly improved, the tissue was neatly arranged, the inflammatory response was alleviated, and the cell morphology tended to be normal. This improvement was dose-dependent; the pathological changes of myocardial tissue in the ASP group and the WSTC+ASP group were also significantly improved.

[0114] The hearts of mice in each group were taken, sliced ​​and subjected to MASSON staining analysis. Fig.13 , the myocardial fibers of the rats in the control group were arranged neatly, with very little interstitial collagen fiber deposition; compared with the control group, the myocardial fibers of the rats in the ISO model group were disordered, with a large number of blue-stained collagen fibers in the interstitium and around the blood vessels, indicating a large area of ​​collagen fiber deposition; compared with the model group, the area of ​​myocardial interstitial collagen deposition in the rats in the WSTC administration group was reduced. The CVF statistical results showed that compared with the control group, the CVF of the rats in the model group was significantly increased (P<0.001); compared with the model group, the CVF of the WSTC dose groups, ASP and WSTC+ASP groups decreased significantly to varying degrees (P<0.001). The results of the present invention show that WSTC can reduce myocardial tissue damage caused by acute myocardial infarction by inhibiting myocardial collagen fiber deposition. The protective effect of the WSTC 50mg / kg group on the myocardium is even better than that of aspirin.

[0115] 2.8 Effect of water-soluble tomato concentrate (WSTC) on cardiomyocyte apoptosis

[0116] Since cardiomyocytes are terminally differentiated cells and do not have the ability to regenerate, ISO-induced cardiomyocyte apoptosis can lead to cell death in the ischemic area and its surrounding areas of the myocardium, ultimately causing irreversible myocardial damage. Fig.14 The results of TUNEL staining showed that compared with the control group, the proportion of apoptotic cardiomyocytes in myocardial tissue of rats in the ISO model group was significantly increased (p<0.001); compared with the model group, the apoptotic rates of cardiomyocytes in the WSTC 25mg / kg group and the WSTC 50mg / kg group were significantly reduced (p<0.01 and p<0.001); the apoptotic rates of cells in the ASP group and the WSTC+ASP group were also significantly decreased (p<0.01 and p<0.05). The results suggest that WSTC can inhibit the apoptosis of cardiomyocytes and improve myocardial damage caused by acute myocardial infarction.

[0117] Example 3 Effect of Water-Soluble Tomato Concentrate (WSTC) Composition on Atherosclerotic Coronary Heart Disease

[0118] Clean-grade healthy male Wistar rats were used as the control group and fed with conventional feed; the model group and the drug group were gavaged with high-fat and vitamin D3 mixed feed to establish a rat atherosclerotic coronary heart disease model, and electrocardiogram examination, physiological signs and cardiac pathological examination were performed using a small animal physiological recorder to confirm that the model group had coronary heart disease. The drug groups were divided into: positive control group (atorvastatin group), sample 1 water-soluble tomato concentrate group (WSTC group), sample 2 ginkgo leaf extract and ginseng extract group (GBR group), sample 3 water-soluble tomato concentrate + ginkgo leaf extract + ginseng extract group (WGR group).

[0119] 3.1 Effects of the test substances on four blood lipid parameters in hyperlipidemia model rats

[0120] After 45 days of oral administration of different doses of the test substance to rats, serum total cholesterol, triglycerides, and low-density lipoprotein increased in the model control group compared with the blank control group (P<0.05), with significant differences; indicating that the model was successfully established. After 45 days of oral administration of different test substances to rats, serum total cholesterol, triglycerides, and low-density lipoprotein decreased in the positive control group and sample 3 intervention group compared with the model control group (P<0.05), with significant differences. The triglyceride level in sample 1 group decreased, and the cholesterol level in sample 2 group decreased (P<0.05), with significant differences. Sample 3 group also produced the effect of lowering LDL-C compared with sample 1 group and sample 2 group, with significant differences p<0.05. In terms of TC index, sample 3 group was also significantly better than sample 1 and sample 2; it can be seen that the combination of water-soluble tomato concentrate with ginkgo leaf extract and ginseng extract played a synergistic role in inhibiting atherosclerosis.

[0121] Table 1 Effects on four blood lipids in hyperlipidemia model rats (mean ± SD, n = 10)

[0122] Group TC TG HDL-C LDL-C Blank control group 2.20±0.27 1.69±0.20 1.08±0.07 0.25±0.04 Model Group 4.30±0.40# 4.53±0.24# 0.54±0.06# 1.22±0.15# Positive control group 3.39±0.36* 3.03±0.22** 0.55±0.08 0.77±0.19** Sample 1 4.13±0.33 4.05±0.25** 0.55±0.05 1.14±0.16 Sample 2 3.89±0.57* 4.69±0.30 0.53±0.07 1.23±0.23 Sample 3 <![CDATA[3.14±0.51* $$ΔΔΔ ]]> 3.90±0.32** 0.53±0.03 <![CDATA[0.98±0.17** $Δ ]]>

[0123] #: There is a significant difference compared with the blank control group *: There is a significant difference compared with the model control group, * indicates p<0.05, ** indicates p<0.01, $ indicates that sample 3 is compared with sample 1, $ indicates p<0.05, $$ indicates p<0.01; Δ indicates that sample 3 is compared with sample 2, Δ indicates p<0.05, ΔΔΔ indicates p<0.001.

[0124] 3.2 Effects of the test substances on inflammatory factors in hyperlipidemia model rats

[0125] After 45 days of oral administration of different doses of the test substance to rats, the levels of TNF-α, IL-1, and CRP in the model control group were significantly increased compared with the blank control group (P<0.05); compared with the model control group, the levels of TNF-α, IL-1, and CRP in the positive control group and sample 1, 2, and 3 intervention groups were significantly decreased (P<0.05).

[0126] Table 2 Effects on inflammation levels in hyperlipidemia model rats (mean ± SD, n = 10)

[0127]

[0128] #: Significantly different from the blank control group *: Significantly different from the model control group, * indicates p<0.05, ** indicates p<0.01

[0129] 3.3 Effects of the test substances on the oxidation level of hyperlipidemia model rats

[0130] After 45 days of oral administration of different doses of the test substance to rats, there was no significant difference in serum MDA, GSH, and GSH-PX between the model control group and the blank control group (P>0.05); compared with the model control group, serum MDA in each dose group was reduced (P<0.05), and the GSH-PX content in sample 3 group increased, with significant differences. Compared with the blank control group, serum SOD in the model control group was reduced (P<0.05), with significant differences. Compared with the model control group, serum SOD in the positive control group, sample 1, sample 2, and sample 3 increased (P<0.05), all with significant differences. Sample 3 group can significantly reduce MDA levels relative to sample 1 and sample 2, and improve the activity of GSH-PX, P<0.01, P<0.001, that is, the combination of water-soluble tomato concentrate with ginkgo leaf extract and ginseng extract can play a synergistic antioxidant role in inhibiting atherosclerosis.

[0131] Table 3 Effects on oxidation levels in hyperlipidemia model rats (mean ± SD, n = 10)

[0132]

[0133]

[0134] #: There is a significant difference compared with the blank control group *: There is a significant difference compared with the model control group, * indicates p<0.05, ** indicates p<0.01; $ is compared between sample 3 and sample 1, $$$ indicates p<0.001; Δ is compared between sample 3 and sample 2, ΔΔΔ indicates p<0.001.

[0135] 3.4 Staining of aortic sections

[0136] Masson staining was performed on the paraffin sections of mouse aorta, and the vascular morphology was observed under an optical microscope to detect the elasticity and collagen fiber content of the aorta. Fig.15 Figure AE, according to the staining results, the model control group (A) contained higher levels of collagen fibers in the aorta. The intervention group (B positive drug group, CE are samples 1-3 respectively) can reduce the deposition of collagen fibers in the vascular wall, especially the improvement of sample 3 group is the most significant.

[0137] In summary, the water-soluble tomato concentrate provided by the present invention can reduce blood lipid levels, play an anti-inflammatory, anti-oxidant, and blood lipid-lowering role in atherosclerosis, and improve and alleviate atherosclerosis. When the water-soluble tomato concentrate is used in combination with ginseng extract and ginkgo leaf extract, it can further play the lipid-lowering (simultaneously reducing serum cholesterol, triglycerides, and LDL), anti-inflammatory, and anti-oxidant effects, and has the ability of synergistic enhancement.

Claims

1. Application of water-soluble tomato concentrate in preparing products for improving heart health.

2. The use according to claim 1, characterized in that: The improvement of heart health is to regulate the myocardial function of the human heart; preferably, the regulation of the myocardial function of the human heart is to increase myocardial blood flow, inhibit myocardial cell damage or apoptosis, reduce myocardial inflammatory factors, immune cell infiltration and / or enhance antioxidant capacity.

3. The use according to claim 1, characterized in that: The improvement of heart health includes preventing and / or treating heart-related diseases; preferably, the heart-related diseases are one or more of stroke, hypertension, heart failure, myocardial infarction, atherosclerosis, and coronary heart disease caused by imbalance of cardiac function.

4. The use according to claim 3, wherein the heart-related disease is heart failure, myocardial infarction or atherosclerotic coronary heart disease.

5. The use according to any one of claims 1 to 4, characterized in that: The product is food, health food or medicine, and the product also includes excipients acceptable to food, health food or medicine.

6. The use according to claim 5, characterized in that: The dosage forms of the product are hard capsules, soft capsules, tablets, powders, gels, lozenges, sustained-release preparations, granule preparations, drops, pills, and liquid preparations.

7. The use according to claim 6, characterized in that: The product also includes one or more of coenzyme Q10, PQQ, NMN, NAD+, turmeric extract, astragalus extract, mulberry bark, scutellaria baicalensis seed extract, Poria cocos extract, ginkgo leaf extract, ginseng extract, American ginseng extract, rhodiola rosea extract, squalene, ganoderma lucidum extract, vitamins, minerals, grape seed extract, bilberry extract, lotus leaf extract, lotus seed core extract, wolfberry extract, red date extract, aspirin, red yeast rice extract, metformin, and ergothioneine.

8. The use according to claim 7, characterized in that: The product also includes aspirin; more preferably, the product also includes ginkgo leaf extract and ginseng extract.

9. The use according to any one of claims 1 or 2, characterized in that: The characteristic components of the water-soluble tomato concentrate are nucleoside compounds, flavonoid compounds and phenolic acid compounds.

10. The use according to claim 9, characterized in that: The water-soluble tomato concentrate is obtained by separating water-soluble components from tomatoes by physical methods, and then concentrating and removing sugar to obtain the water-soluble components.

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