Active substance for strengthening cardiovascular functions of dogs and cats and preparation method and application of soft capsule

Abstract

The invention discloses an active substance for strengthening cardiovascular functions of dogs and cats and a preparation method and application of soft capsules, and relates to the technical field of animal health care products. The composition comprises components such as kelp powder extract, coix seed extract, Omega-3 fatty acid, coenzyme Q10, L-carnitine, vitamin B6, vitamin E and the like, and the components jointly act on proliferation, metabolism and transcription processes of vascular endothelial cells of dogs and cats. Through in-vitro metabonomics and transcriptomics analysis, the composition has the effects of promoting proliferation of vascular endothelial cells of dogs and cats, increasing cell elasticity, reducing the risk of thrombus and sclerosis and the like. In addition, the invention also provides application of the composition in preparation of food for strengthening cardiovascular functions of dogs and cats. Compared with the prior art, the composition provided by the invention can provide an effective health-care strategy and can remarkably promote cardiovascular functions of dogs and cats.

Description

Technical Field

[0001] The invention belongs to the technical field of animal health products, and in particular relates to an active substance for strengthening the cardiovascular function of dogs and cats and a preparation method and application of its soft capsule. Background Art

[0002] With the popularity of pet keeping, people are paying more and more attention to the health of dogs and cats, especially the cardiovascular health. Cardiovascular diseases are common in pets, including heart disease, hypertension, atherosclerosis and other problems, which not only affect the quality of life of pets, but may also shorten their lifespan. Therefore, it is particularly important to maintain or improve the cardiovascular health of pets through a reasonable diet and supplements.

[0003] At present, there are many health supplements on the market that use natto powder, red yeast rice powder, collagen, cysteine, glutathione, kudzu root, cyclocarya paliurus, coenzyme Q10, etc. to strengthen cardiovascular function. For example, CN 110051826A discloses a composition for improving cardiovascular function, which includes natto powder, red yeast rice powder, collagen, taurine, γ-aminobutyric acid, salt algae powder, etc. Similarly, CN 112450436A discloses a food composition for improving cardiovascular function and its application, which includes L-cysteine, water-soluble cellulose, glutathione, SOD enzyme, phytosterols, soybean lecithin, soybean oligosaccharides, etc. CN115708533A discloses a functional tea for preventing and improving cardiovascular and cerebrovascular diseases and its preparation method, wherein the dry raw materials of the functional tea include kudzu root, cyclocarya paliurus, thorn tea, mulberry leaf, grape seed, Hovenia dulcis, sea buckthorn fruit, and golden camellia. In addition, CN113397058A discloses a dog food for improving cardiovascular function, the main ingredients of which include L-carnitine, taurine, rosemary extract, methionine, etc. However, there is no clear conclusion on the pathological mechanism and treatment strategy of cardiovascular diseases in dogs and cats based on these functional ingredients. Although existing studies include some ingredients such as natto powder, L-cysteine, L-carnitine, etc., other key biological pathways have not been fully considered. For example, important aspects such as metabolic changes in vascular endothelial cells of dogs and cats and abnormal intracellular signal transduction have not been fully studied. Therefore, existing treatment strategies have certain limitations in the basic pathological mechanisms of cardiovascular diseases in dogs and cats.

[0004] Due to the limitations of current treatment methods and the lack of in-depth understanding of pathological mechanisms, the present invention conceived a new cardiovascular health care strategy for dogs and cats, the core of which is the precise regulation of nutritional metabolites. After conducting in-depth research on the metabolic and transcriptional mechanisms of vascular endothelial cells at the cellular level, we successfully identified several key biological metabolic pathways. The metabolite regulation in these pathways can provide a more complete solution for the treatment of cardiovascular diseases in dogs and cats. Based on this, we have also developed a new formula. Summary of the invention

[0005] In view of this, the present invention aims to propose an active substance for strengthening the cardiovascular function of dogs and cats and a method for preparing and applying its soft capsule, and to study the changes of the active ingredients in increasing the elasticity of vascular endothelial cells, reducing cell sclerosis and thrombus formation. Subsequently, the metabolomics and transcriptomics of cat vascular endothelial cells were determined by high performance liquid chromatography tandem mass spectrometry (HPLC-MS / MS) and RNA sequencing (RNA-seq) technology. The metabolomics data showed that the active substance could significantly affect amino acid metabolism, lipid metabolism, etc. Differential metabolite screening determined the changes in the content of key metabolites in vascular endothelial cells, and caused significant abnormalities in various related key metabolic pathways, including propionic acid metabolism, sulfur metabolism, fluid shear stress and atherosclerosis, folic acid biosynthesis, linolenic acid metabolism, oxidative phosphorylation, etc. Transcriptome data showed that the active substance affected vascular endothelial cells through pathways such as cytoskeleton protein binding, ubiquitin protein transferase activity, Ras GTPase binding and enzyme binding.

[0006] Based on the above findings, the present invention provides an active substance for strengthening the cardiovascular function of dogs and cats, which is composed of the following components by weight:

[0007]

[0008] Preferably, the above active substance for strengthening the cardiovascular function of dogs and cats is composed of the following components by weight:

[0009]

[0010] The present invention also provides a method for preparing a soft capsule containing the active substance for strengthening the cardiovascular function of dogs and cats, which is carried out according to the following steps:

[0011] (1) Weighing and preparing materials: Accurately weigh kelp powder extract, coix seed extract, omega-3 fatty acids, coenzyme Q10, L-carnitine, vitamin B6, vitamin E, chicken powder, soybean powder, and chicken fat and set aside.

[0012] (2) Stirring and sieving: Stir and mix the kelp powder extract, coix seed extract, vitamin B6, vitamin E, chicken powder and soybean powder, and pass the materials through an 80-mesh sieve.

[0013] (3) Mixing: Add omega-3 fatty acids, coenzyme Q10, L-carnitine, and chicken fat to the stirred and sieved mixture and mix thoroughly;

[0014] (4) Granulation: The mixed material is sent to a low-temperature cold-pressing granulator, and soft granules are extruded through the low-temperature cold-pressing granulator.

[0015] (5) Packaging: Pack the soft capsules into finished products and seal them.

[0016] Combined with the metabolism and transcription mechanism of cat vascular endothelial cells, the effects of each raw material in the present invention are reported as follows:

[0017] Kelp powder extract and coix seed extract: can enhance the activity of vascular endothelial cells in dogs and cats, promote the production of short-chain fatty acids, activate specific signaling pathways and affect the expression of metabolism-related genes.

[0018] Omega-3 fatty acids: can change the composition of cell membranes, thereby affecting the signal conduction process. It can improve heart function by lowering blood lipid levels.

[0019] Coenzyme Q10: Involved in ATP production. Its antioxidant properties help scavenge free radicals and reduce DNA damage. At the transcriptional level, CoQ10 supplementation may upregulate the expression of genes involved in the antioxidant defense system.

[0020] L-carnitine: It is directly involved in the process of fatty acid transport to mitochondria for oxidation and decomposition, which not only improves energy utilization, but also reduces the toxicity caused by the accumulation of free fatty acids. At the molecular level, the presence of L-carnitine may promote the expression of enzymes related to fatty acid oxidation.

[0021] Vitamin B6, Vitamin E: Vitamin B6 participates in multiple metabolic pathways as a coenzyme, including amino acid metabolism and carbon monoxide transfer; Vitamin E prevents lipid peroxidation and protects the integrity of cell membrane structure through its antioxidant effect.

[0022] Compared with the prior art, the present invention has the following advantages:

[0023] 1. Different from the prior art used in the fields of cosmetics and medicine, the present invention focuses on the field of animal food. The present invention is aimed at the cellular level research of dogs and cats, and on this basis, the metabolic and transcriptional changes of vascular endothelial cells of dogs and cats are deeply explored, and the mechanism of action in fat metabolism, amino acid metabolism, etc. is found. By introducing key ingredients such as L-carnitine and vitamin B6, the present invention provides comprehensive metabolic support for the process of increasing cell elasticity, reducing hardening and thrombosis of vascular endothelial cells of dogs and cats, thereby achieving the strengthening of cardiovascular function of dogs and cats.

[0024] 2. The active substances of the present invention effectively regulate key biochemical pathways in vascular endothelial cells of dogs and cats by supplementing a variety of nutritional metabolic regulators. This comprehensive nutritional support method helps to fundamentally improve the elasticity of vascular endothelial cells of dogs and cats and reduce the risk of sclerosis and thrombosis, surpassing the limitations of traditional supplements.

[0025] These improvements ensure that the present invention has significant advantages in providing proliferation of canine and feline vascular endothelial cells, and show obvious innovation and improvement compared with the prior art. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the accompanying drawings:

[0027] Cat vascular endothelial cells were grouped as follows: BC was the blank control group; H2O2 was the 0.2mmol / L H2O2 treatment group; KPE+H2O2 was the 1μg / mL KPE+0.2mmol / L H2O2 treatment intervention group; CSE+H2O2 was the 1μg / mL CSE+0.2mmol / L H2O2 treatment intervention group; oleic acid (OA) was the 0.2mmol / L OA treatment group; KPE+OA was the 1μg / mL KPE+0.2mmol / L OA treatment intervention group; CSE+OA was the 1μg / mL CSE+0.2mmol / L OA treatment intervention group.

[0028] Canine vascular endothelial cells were grouped as follows: BC was the blank control group; H2O2 was the 1mmol / L H2O2 treatment group; KPE+H2O2 was the 1μg / mL KPE+1mmol / L H2O2 treatment intervention group; CSE+H2O2 was the 1μg / mL CSE+1mmol / L H2O2 treatment intervention group; OA was the 0.3mmol / L OA treatment group; KPE+OA was the 1μg / mL KPE+0.3mmol / L OA treatment intervention group; CSE+OA was the 1μg / mL CSE+0.3mmol / L OA treatment intervention group.

[0029] Figure 1 Effects of KPE and CSE intervention on oxidative stress indicators of vascular endothelial cells. (A) Lactate dehydrogenase (LDH) activity of cat vascular endothelial cells; (B) Malondialdehyde (MDA) content of cat vascular endothelial cells; (C) Lactate dehydrogenase (LDH) activity of dog vascular endothelial cells; (D) Malondialdehyde (MDA) content of dog vascular endothelial cells.

[0030] Figure 2 Effects of KPE and CSE intervention on NO content in vascular endothelial cells. (A) NO content in cat vascular endothelial cells; (B) NO content in dog vascular endothelial cells.

[0031] Figure 3 Effects of KPE and CSE intervention on endothelial dysfunction-related indicators in vascular endothelial cells. (A) Contents of sVCAM-1, CD62P, and vWF in cat vascular endothelial cells; (B) Contents of sVCAM-1, CD62P, and vWF in dog vascular endothelial cells.

[0032] Figure 4Effects of KPE and CSE intervention on lipid-related indexes in vascular endothelial cells. (A) TG, LDL-C, and HDL-C levels in cat vascular endothelial cells; (B) TG, LDL-C, and HDL-C levels in dog vascular endothelial cells.

[0033] Figure 5 Effects of KPE and CSE intervention on cell elasticity in vascular endothelial cells. (A) Fluorescence image of cell elasticity of cat vascular endothelial cells; (B) Fluorescence image of cell elasticity of cat vascular endothelial cells.

[0034] Figure 6 Volcano plots and cluster heat maps of differentially expressed metabolites between groups. (AC) Volcano plots of differential metabolites between groups; (D) Cluster heat maps of differential metabolites between groups.

[0035] Figure 7 Volcano plots and cluster heat maps of differentially expressed genes between groups. (AC) Volcano plots of differentially expressed genes between groups; (D) Cluster heat maps of differentially expressed genes between groups. DETAILED DESCRIPTION

[0036] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0037] The present invention will be described in detail below with reference to the accompanying drawings and in conjunction with embodiments.

[0038] The kelp powder extract and coix seed extract of the present invention are purchased from Shaanxi Xintianyu Biotechnology Co., Ltd.

[0039] Example 1

[0040] Oxidative stress index detection

[0041] Feline (canine) vascular endothelial cells were cultured in M-3 medium containing 10% fetal bovine serum and 1% double antibody, placed in a 37°C, 5% CO2 incubator, and cell passage and experiment were performed when the cells grew to 80% confluence. The cells were digested with trypsin, and the culture medium was added to adjust the cell concentration to 1×10 5 cells / mL, 1mL per well was inoculated in a 6-well plate, and placed in an incubator for 24h, and then H2O2 (cat: 0.2mmol / L; dog: 1mmol / L) was added for 2h. The cells were washed three times with phosphate buffer (PBS, pH=7.4), and 1mL KPE (1μg / mL) and 1mL CSE (1μg / mL) solutions were added as the test group; only H2O2 was used as the negative control; the six-well plate without treatment was the blank control. After 24h of continuous culture, the cells were lysed with lysis buffer, and the indicators were detected using LDH and MDA kits.

[0042] KPE and CSE can effectively alleviate the oxidative stress level of cat (dog) vascular endothelial cells induced by H2O2. Figure 1 As shown. Compared with the BC group, the extracellular LDH leakage of cells in the H2O2 group increased, and the intracellular MDA level increased. This shows that cat (dog) vascular endothelial cells induced by H2O2 produced an oxidative stress response. Compared with the H2O2 group, under the intervention of 1μg / mL KPE and 1μg / mL CSE, the cellular LDH leakage and MDA level were inhibited, indicating that KPE and CSE can effectively slow down the oxidative stress response of cells induced by H2O2 and enhance the antioxidant capacity.

[0043] Example 2

[0044] NO content detection

[0045] The cells were treated according to the method of Example 1, cultured for 24 hours, centrifuged at 2000 r / min for 10 minutes, and the supernatant was collected. The release of inflammatory mediator NO from the cells was detected by nitrate reductase method. The operation was carried out according to the instructions of the NO detection kit, and the absorbance value at a wavelength of 540 nm was measured using an ELISA instrument.

[0046] KPE and CSE can effectively reduce the release of NO in cat (dog) vascular endothelial cells induced by H2O2. Figure 2 As shown in the figure, compared with the BC group, the H2O2 group had an increased NO release, resulting in an inflammatory response. Compared with the H2O2 group, the NO content in the cells was significantly reduced under the intervention of 1μg / mL KPE and 1μg / mL CSE. This indicates that KPE and CSE inhibited H2O2-induced oxidative stress and inflammatory response by regulating the NO production pathway, thereby protecting cells from further damage.

[0047] Example 3

[0048] Endothelial dysfunction related indexes

[0049] The cells were treated according to the method of Example 1, and the supernatant was collected after culturing for 24 hours. The concentrations of vascular cell adhesion molecule-1 (sVCAM-1), P-selectin (CD62P) and von Willebrand factor (vWF) were detected by enzyme-linked immunosorbent assay (ELISA), and the operation was carried out according to the instructions of the ELISA detection kit, and the absorbance value at a wavelength of 450 nm was measured using an enzyme marker.

[0050] KPE and CSE can effectively slow down the damage and activation of cells induced by H2O2. Figure 3As shown in the figure, compared with the BC group, the levels of sVCAM-1, CD62P, and vWF in the H2O2 group were significantly increased. This indicates that after H2O2 induction, vascular endothelial cells underwent significant damage, which may lead to endothelial dysfunction and increase the risk of sclerosis and thrombosis. Compared with the H2O22 group, the levels of sVCAM-1, CD62P, and vWF were significantly reduced under the intervention of 1μg / mL KPE and 1μg / mL CSE. This indicates that KPE and CSE can effectively slow down H2O2-induced vascular endothelial cell damage and activation, thereby reducing the risk of thrombosis.

[0051] Example 4

[0052] Determination of blood lipid related indicators

[0053] Feline (canine) vascular endothelial cells were cultured in M-3 medium containing 10% fetal bovine serum and 1% double antibody, placed in a 37°C, 5% CO2 incubator, and cell passage and experiment were performed when the cells grew to 80% confluence. The cells were digested with trypsin, and the culture medium was added to adjust the cell concentration to 1×10 5 cells / mL, 1mL per well was inoculated in a 6-well plate, and after culturing in an incubator for 24h, OA (cat: 0.2mmol / L; dog: 0.3mmol / L) was added for 24h. Washed with PBS three times, 1mL KPE (1μg / mL) and 1mL CSE (1μg / mL) solution were added as the experimental group; only OA was used as the negative control; the six-well plate without treatment was the blank control. After culturing for another 24h, the cells were lysed with lysis buffer, and the indicators were detected using TG, LDL-C, and HDL-C kits.

[0054] KPE and CSE can effectively reduce OA-induced lipid accumulation in cat vascular endothelial cells. Figure 4 As shown in the results, compared with the BC group, the TG and LDL-C levels in the OA group were significantly increased, and the HDL-C content was decreased. This indicates that after OA induction, the lipid levels in vascular endothelial cells changed, which may lead to endothelial dysfunction and increase the risk of thrombosis. Compared with the OA group, under the intervention of 1μg / mL KPE and 1μg / mL CSE, the TG and LDL-C levels decreased, and the HDL-C content increased. This indicates that KPE and CSE can effectively reduce the lipid accumulation in vascular endothelial cells induced by OA, thereby reducing the risk of thrombosis.

[0055] Example 5

[0056] Cell elasticity assay

[0057] Treat cells according to the method of Example 1 and culture for 24 hours. Rinse twice with PBS, fix with 4% paraformaldehyde for 20 minutes, and then rinse with 0.1% TritonX-100-PBS solution for 2-4 times, 5 minutes each time. Mix 1% blocking protein (BSA) and Actin-Tracker Red-555 at a ratio of 40:1, add 200 μL to each well and incubate in the dark for 30 minutes. Finally, rinse with 0.1% TritonX-100-PBS solution for 2-4 times, 5 minutes each time. Observe under a fluorescence microscope.

[0058] KPE and CSE can effectively reduce the damage of F-actin skeleton in vascular endothelial cells induced by H2O2. Figure 5 As shown in the figure, compared with the BC group, the F-actin distribution after H2O2 treatment was broken. The fluorescence intensity was weakened, reflecting obvious cytoskeletal damage. This indicates that vascular endothelial cells experienced significant cell damage after H2O2 induction. Compared with the H2O2 group, the F-actin skeleton fracture state of vascular endothelial cells treated with KPE and CSE was improved, the fluorescence intensity was enhanced, and the cell morphology was more complete. This indicates that KPE and CSE help protect the cytoskeletal structure.

[0059] Example 6

[0060] In order to gain a deeper understanding of the effects of KPE and CSE on vascular endothelial cells, cat vascular endothelial cells were used as an example to conduct a detailed metabolomics analysis of cat vascular endothelial cells using high performance liquid chromatography tandem mass spectrometry (HPLC-MS / MS). The cells were treated according to the method in Example 1 and cultured for 24 hours. The cells treated with H2O2 were used as the control group, and the cells treated with KPE were used as the experimental group.

[0061] Figure 6The volcano plot and cluster heat map of differential metabolites between the groups. The results showed that compared with the BC group, 14 metabolites were upregulated and 23 metabolites were downregulated in the H2O2-treated group; 15 metabolites were upregulated and 131 metabolites were downregulated in the KPE-treated group. Compared with the H2O2-treated group, 19 metabolites were upregulated and 76 metabolites were downregulated in the KPE-treated group. H2O2-induced vascular endothelial cell injury is caused by upregulating L-octanoylcarnitine and aminopyrazone, and downregulating glutamyllysine, acetaminophen sulfate, 7-N-glucoside and other metabolites. These metabolites are mainly involved in pathways such as fluid shear stress and atherosclerosis, anti-folate resistance, and folate biosynthesis. After KPE treatment, the metabolite changes were significantly different from those under H2O2 induction. The enrichment results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) database showed that KPE is involved in oxygen pathways such as propionate metabolism, sulfur metabolism, oxidative phosphorylation, and insulin resistance. These pathways play an important role in cellular amino acid metabolism and lipid metabolism. Therefore, KPE alleviates H2O2-induced cell damage by affecting the content of metabolites in these pathways.

[0062] Example 7

[0063] In order to gain a deeper understanding of the effects of KPE and CSE on vascular endothelial cells, cat vascular endothelial cells were used as an example, and transcriptomic sequencing of cell RNA was performed based on the Illumina sequencing platform. The cells were treated according to the method in Example 1 and cultured for 24 hours. The cells treated with H2O2 were used as the control group, and the cells treated with KPE were used as the experimental group. An equal volume of samples was taken from each experimental sample and mixed as a quality control group. The RNA of each group was extracted using standard methods, the library was constructed, and the sequencing was performed on the machine. The differential expression analysis between the two comparison combinations was performed using DESeq2 software (1.20.0).

[0064] Figure 7The volcano plot and cluster heat map of differentially expressed genes between the groups. The results showed that compared with the BC group, 255 genes were upregulated and 389 genes were downregulated in the H2O2 treatment group; 1046 genes were upregulated and 1887 genes were downregulated in the KPE treatment group. Compared with the H2O2 treatment group, 306 genes were upregulated and 998 genes were downregulated in the KPE treatment group. H2O2-induced endothelial cell injury is caused by downregulating enzyme activity or protein expression related to redox processes and lipid metabolism. These genes are mainly involved in pathways such as DNA replication, redox processes, lipid metabolism processes, oxidoreductase activity, copper ion binding, iron ion binding, and ribonucleic acid binding. After KPE treatment, the changes in gene expression were significantly different from those under H2O2 induction. The results of annotation enrichment on the differentially expressed gene function (GO) database showed that items such as cytoskeletal protein binding, ubiquitin protein transferase activity, ubiquitin-like protein transferase activity, Ras GTPase binding, small GTPase binding, GTPase binding, enzyme binding, non-membrane-bound organelles, and intracellular non-membrane-bound organelles were significantly enriched. The KEGG database enrichment results showed that KPE was involved in pathways such as oxidative phosphorylation, Parkinson's disease, neurodegenerative diseases, diabetic cardiomyopathy, ribosomes, proteasomes, and myocardial contraction. These pathways play an important role in cell movement, differentiation, proliferation, and other processes. Therefore, KPE alleviates H2O2-induced cell damage by affecting the expression of genes or enzymes in these pathways.

[0065] Examples 8-10

[0066] Examples 8 to 10 Soft capsules were prepared according to the formulations shown in Table 1 in the following manner:

[0067] (1) Weighing and preparing materials: accurately weigh kelp powder extract, coix seed extract, omega-3 fatty acids, coenzyme Q10, L-carnitine, vitamin B6, vitamin E, chicken meal, soybean meal, and chicken fat for later use;

[0068] (2) Stirring and sieving: stir and mix the kelp powder extract, coix seed extract, chicken powder, and soybean powder, and pass the materials through an 80-mesh sieve;

[0069] (3) Mixing: Add omega-3 fatty acids, coenzyme Q10, L-carnitine, vitamin B6, vitamin E and chicken fat to the stirred and sieved mixture and mix thoroughly;

[0070] (4) Granulation: The mixed material is sent to a low-temperature cold-pressing granulator, and soft granules are extruded through the low-temperature cold-pressing granulator;

[0071] (5) Packaging: Pack the soft capsules into finished products and seal them.

[0072] Comparative Example 1-2

[0073] Comparative Example 1-2 was prepared according to the formula shown in Table 1. The preparation process involved weighing and preparing materials, granulation and drying, total mixing and packaging, which were the same as those of Examples 8 to 10. The numerical units in Table 1 are grams. Each capsule weighed about 500±10 mg, and a total of 200 soft capsules were prepared.

[0074] Table 1 Formulas used in various embodiments and comparative examples

[0075]

[0076] Test example

[0077] 1. Test methods

[0078] (1) Experimental animals and groups: Twenty cats (dogs) with weak cardiovascular function were selected and randomly divided into five groups, with four cats in each group.

[0079] 2. Intervention plan: Groups 1-3 are experimental groups 1-3, fed with a basic diet and soft capsules prepared in 8-10 daily. Groups 4-5 are control groups, fed with a basic diet and soft capsules prepared in 1-2 daily. The dosage of each group is 2 capsules each time, once a day, for a feeding period of 3 months. The activity was observed, and after 3 months, blood was collected to test the total cholesterol, triglyceride, and high-density lipoprotein levels, and Doppler ultrasound was used to observe the thickness of the blood vessel wall. Experimental results

[0080] The effects of each group of cats and dogs are shown in Table 2 and Table 3. Compared with the control group, feeding Examples 1-3 can reduce the total cholesterol and triglyceride content of cats (dogs), increase the high-density lipoprotein content, and thin the blood vessel wall thickness.

[0081] Table 2 Results of cardiovascular function indexes in each group of cats

[0082]

[0083] Table 3 Results of cardiovascular function indexes in each group of dogs

[0084]

[0085] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An active substance for strengthening the cardiovascular function of dogs and cats, characterized in that It is composed of the following components by weight:

2. The active substance for strengthening cardiovascular function of dogs and cats according to claim 1, characterized in that It is composed of the following components by weight:

3. A method for preparing a soft capsule containing the above-mentioned active substance for strengthening the cardiovascular function of dogs and cats according to claim 1 or 2, characterized in that Follow the steps below: (1) Weighing and preparing materials: Accurately weigh kelp powder extract, coix seed extract, omega-3 fatty acids, coenzyme Q10, L-carnitine, vitamin B6, vitamin E, chicken powder, soybean powder, and chicken fat and set aside. (2) Stirring and sieving: Stir and mix the kelp powder extract, coix seed extract, vitamin B6, vitamin E, chicken powder and soybean powder, and pass the materials through an 80-mesh sieve. (3) Mixing: Add omega-3 fatty acids, coenzyme Q10, L-carnitine, and chicken fat to the stirred and sieved mixture and mix thoroughly; (4) Granulation: The mixed material is sent to a low-temperature cold-pressing granulator, and soft granules are extruded through the low-temperature cold-pressing granulator. (5) Packaging: Pack the soft capsules into finished products and seal them.

4. Use of the active substance according to any one of claims 1 to 2 in food for strengthening the cardiovascular function of dogs and cats.

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