Use of seabuckthorn seed oil in the preparation of a drug for preventing and / or treating cardiovascular diseases

Abstract

The application provides application of seabuckthorn seed oil in preparation of a medicine for preventing and / or treating cardiovascular diseases, and belongs to the technical field of biological medicine preparation.The application provides application of seabuckthorn seed oil in preparation of a medicine for preventing and / or treating cardiovascular diseases.The application takes a HHcy-induced myocardial fibrosis model of mice as an experimental object to evaluate the drug efficacy of seabuckthorn seed oil, and the results show that the seabuckthorn seed oil can effectively improve the myocardial dysfunction of mice induced by homocysteine, and reduce the degree of myocardial fibrosis; and play an anti-fibrosis role by inhibiting the expression of fibrosis markers such as alpha-SMA, type I collagen, type III collagen and the like in myocardial tissue. It can be seen that the application provides a new choice for preparation or development of a medicine for preventing and assisting in treating HHcy-related cardiovascular diseases, and also widens the medical use of seabuckthorn seed oil.

Description

Technical Field

[0001] This application belongs to the field of biopharmaceutical preparation technology, specifically relating to the application of sea buckthorn seed oil in the preparation of drugs for the prevention and / or treatment of cardiovascular diseases. Background Technology

[0002] Hyperhomocysteinemia (HHcy) is a condition characterized by elevated homocysteine ​​levels in the blood. The normal reference range is 5–15 μmol / L. If the level is normal, medication is generally not required. Homocysteine ​​levels >15 μmol / L are considered hyperhomocysteinemia. Hyperhomocysteinemia is associated with various cardiovascular diseases. For example, it can lead to increased myocardial thickness, increased myocardial microvessels and collagen content, resulting in decreased diastolic function and compliance, potentially promoting severe myocardial fibrosis and exacerbating left ventricular structural and functional impairments.

[0003] Currently, the mainstream treatment regimen is folic acid combined with B vitamins and / or betaine, which has a clear effect on reducing homocysteine ​​(Hcy) and preventing fibrosis. Recent research reports that H2S can reduce HHcy-induced myocardial fibrosis. It is evident that the types of drugs or regimens for treating homocysteine-induced myocardial fibrosis are very limited, mainly relying on basic interventions and lacking highly specific targeted therapies. Furthermore, existing treatments are all post-disease interventions, and there are no preventative treatment programs for high-risk HHcy populations, failing to block the initiation of myocardial fibrosis at its source. Summary of the Invention

[0004] This invention provides a new pharmaceutical use for sea buckthorn seed oil, namely, its use in the preparation of medicines for the prevention and / or treatment of cardiovascular diseases.

[0005] This invention provides the use of sea buckthorn seed oil in the preparation of medicaments for the prevention and / or treatment of cardiovascular diseases.

[0006] Preferably, the cardiovascular disease includes cardiovascular disease associated with hyperhomocysteinemia.

[0007] Preferably, the hyperhomocysteinemia-related cardiovascular disease includes hyperhomocysteinemia-induced myocardial fibrosis.

[0008] Preferred methods for preventing cardiovascular disease associated with hyperhomocysteinemia include preventing or delaying the onset of myocardial fibrosis induced by hyperhomocysteinemia.

[0009] Preferably, the drug includes a drug that improves at least one of the following: myocardial structure, degree of myocardial fibrosis, and myocardial collagen deposition in hyperhomocysteinemia.

[0010] Preferably, the drug comprises a solid dosage form and / or a liquid dosage form.

[0011] Preferably, the solid dosage form includes at least one of the following: tablets, soft capsules, powders, granules, powders, and pellets.

[0012] Preferably, the liquid formulation includes at least one of the following: oral solution, oral suspension, oral emulsion, and syrup.

[0013] Preferably, the drug further includes pharmaceutically acceptable excipients.

[0014] Preferably, the sea buckthorn seed oil content in the drug is 5% or more by mass.

[0015] This invention provides the application of sea buckthorn seed oil in the preparation of drugs for the prevention and / or treatment of cardiovascular diseases. To explore the intervention effect of sea buckthorn seed oil on HHcy-induced myocardial fibrosis and evaluate its efficacy in inhibiting fibrosis, antioxidation, anti-inflammation, and improving microcirculation, an HHcy-induced mouse myocardial fibrosis model was established by feeding mice with a 2% high-methionine diet. This invention set up a control group, a model group, a sea buckthorn seed oil treatment group, a sea buckthorn seed oil prevention group, and a folic acid-positive control group. Cardiac function indicators such as left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were detected by echocardiography; myocardial collagen deposition was assessed by Masson staining. The results showed that the sea buckthorn seed oil could effectively improve hyperhomocysteinemia-induced cardiac dysfunction in mice and reduce the degree of myocardial fibrosis. Further molecular detection and transcriptomic analysis confirmed that the anti-fibrotic effect of sea buckthorn seed oil is related to its inhibition of the expression of fibrosis markers such as α-SMA, type I collagen, and type III collagen in myocardial tissue, further revealing the mechanism by which sea buckthorn seed oil improves HHcy-induced myocardial fibrosis. Therefore, this invention provides a new option for the preparation or development of drugs for the prevention and adjunctive treatment of HHcy-related cardiovascular diseases, and also broadens the medicinal uses of sea buckthorn seed oil. Attached Figure Description

[0016] Figure 1 Results of successful HHcy mouse model establishment: A shows the gross appearance (top) and gross morphology of the heart (bottom) of mice in each group; B shows the H&E staining results of the myocardium; scale bar is 50 μm; C shows the serum homocysteine ​​(Hcy) concentration of mice in each group. P <0.01 vs control group; D represents the results of MASSON staining of the myocardium, scale bar is 50 μm; Figure 2To illustrate the effect of sea buckthorn seed oil on myocardial function in Hhcy mice, A shows cardiac ultrasound images (including two-dimensional ultrasound and M-mode ultrasound), and B shows the results of ejection fraction (EF), fractional shortening (FS), cardiac output (CO), and end-systolic volume measurements. Figure 3 The results of relevant indicators in mice are shown in Figure A. The gross appearance of mice in each group (top) and the gross morphology of the heart (bottom) are shown in Figure B. The serum homocysteine ​​(Hcy) concentration of mice in each group is shown in Figure B. P <0.05 vs. control group; # P <0.05 vs model group; C is the ratio of heart weight to body weight of mice in each group; D is the blood glucose value of mice at different ages in each group; E is the growth curve of mice at different ages and body weight in each group. Figure 4 The effects of sea buckthorn seed oil on myocardial structure and fibrosis in HHcy mice are shown in Figure A, where H&E staining results are shown in scale bar 50 μm; and Masson staining results are shown in scale bar 100 μm, where blue represents collagen fibers. Figure 5 To illustrate the effect of sea buckthorn seed oil on improving myocardial collagen deposition in HHcy mice, A shows the immunohistochemical staining results of type I collagen (Col I), and B shows the immunohistochemical staining results of type III collagen (Col III); scale bar is 100 μm. Figure 6 The transcriptome sequencing results are shown below. A is a bar chart showing differentially expressed genes between the model group and the control group (left), and between the sea buckthorn treatment group and the model group (right), with red indicating upregulated genes and blue indicating downregulated genes. B is a Venn diagram representing key differentially expressed genes reversing sea buckthorn reversal. C is a heatmap of differentially expressed genes from the intersection. D is a bar chart of GO functional enrichment analysis of differentially expressed genes from the intersection. E is a bubble chart of KEGG pathway enrichment analysis of differentially expressed genes from the intersection, with the size of the dots representing the number of genes and the color representing -log10( P value); Figure 7 The results are for α-SMA detection. A is immunofluorescence staining, where green fluorescence represents positive expression of α-SMA; B is immunohistochemical staining, where brown-yellow staining represents positive expression of α-SMA; the groups are as follows: Control (control group), Model (model group), FA (folic acid intervention group), and SSO (sea buckthorn seed oil intervention group). Detailed Implementation

[0017] This invention provides the use of sea buckthorn seed oil in the preparation of medicaments for the prevention and / or treatment of cardiovascular diseases.

[0018] In this invention, the sea buckthorn seed oil is obtained from sea buckthorn (Hippophae rhamnoides), a plant belonging to the genus Hippophae in the family Elaeagnaceae. Hippophae rhamnoides Sea buckthorn seed oil is a natural oil extracted from the seeds of sea buckthorn. It is a brownish-yellow to brownish-red transparent oily liquid and is a highly concentrated product of the bioactive components of sea buckthorn seeds. In this invention, the preferred method for preparing the sea buckthorn seed oil includes the following steps: after pressing the sea buckthorn seeds, separating the crude oil and seed meal; subjecting the seed meal to supercritical CO2 extraction to separate the oil fraction; combining the oil fraction and crude oil, removing impurities, and obtaining sea buckthorn seed oil. The sea buckthorn seeds preferably undergo pretreatment before pressing. The pretreatment method involves passing the dried sea buckthorn seeds through a gravity sieve and air classification to remove impurities, then mechanically separating the shell from the kernel to obtain sea buckthorn seeds. The sea buckthorn seeds are then dried at a low temperature below 50°C, controlling the moisture content to between 5% and 7%. The pressing temperature is preferably below 60°C, but can be 30-55°C or 40-50°C. The pressing pressure is preferably 20-35 MPa, but can be 22-33 MPa, 25-30 MPa, or 27 MPa. The purpose of low-temperature pressing is to avoid the damage of unsaturated fatty acids and heat-sensitive active ingredients (such as tocopherols and sterols) caused by high temperatures. The extraction pressure of the supercritical CO2 extraction is preferably 25-35 MPa, but can be 28-32 MPa or 30 MPa. The extraction temperature is preferably 35-45℃, but can be 38-42℃. The CO2 flow rate is preferably 25-35 L / h, but can be 28-32 L / h or 30 L / h. The extraction time is preferably 2-3 hours, but can be 2.5 hours. Supercritical CO2 extraction selectively extracts high-purity oil fractions rich in palmitic acid, linoleic acid, vitamin E, and sterols through gradient pressure reduction in a separator. The volume ratio of the oil fraction to crude oil is preferably 2-4:6-8, but can also be 3:7. The impurity removal method preferably involves treating the mixed oil under low-temperature (≤80℃) vacuum degumming, deacidification, decolorization, dewaxing, and dehydration under an inert gas (such as nitrogen) protection. The decolorization matrix is ​​preferably a composite adsorbent of activated clay and attapulgite (volume ratio 3:1). The decolorization is preferably carried out under a vacuum of ≥0.095 MPa to effectively remove impurities while maximizing the retention of natural pigments and active substances. The impurity removal also includes nanofiltration. The filter membrane used for nanofiltration is preferably 0.22 μm. The final oil yield (based on seed kernels) of the method is ≥85%, and the total unsaturated fatty acid content in the sea buckthorn seed oil is ≥85%, of which palmitoleic acid (C16:1, Omega-7) content is ≥30%, and α-tocopherol content is ≥200 mg / 100g.

[0019] In this invention, the cardiovascular disease preferably includes cardiovascular disease associated with hyperhomocysteinemia. The cardiovascular disease associated with hyperhomocysteinemia preferably includes myocardial fibrosis induced by hyperhomocysteinemia. Hyperhomocysteinemia is defined as a homocysteine ​​level >15 μmol / L in the patient. Prevention of cardiovascular disease associated with hyperhomocysteinemia includes preventing or delaying the onset of myocardial fibrosis induced by hyperhomocysteinemia. In this embodiment of the invention, a mouse model of myocardial fibrosis induced by hyperhomocysteinemia is used as the experimental subject. The hyperhomocysteinemia is preferably induced by a methionine-rich diet. The methionine-rich diet contains 2% methionine by mass. The induction period is 8 weeks. The mouse model of myocardial fibrosis induced by hyperhomocysteinemia is evaluated by detecting serum Hcy levels, H&E staining, and Masson staining to assess myocardial pathological changes and the degree of collagen deposition, and to evaluate whether the disease model has been successfully established.

[0020] In this invention, the drug preferably includes a drug that improves at least one of the following: myocardial structure, degree of myocardial fibrosis, and myocardial collagen deposition in hyperhomocysteinemia. In embodiments of this invention, echocardiography was used to detect cardiac function indicators such as left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) to assess the effect of sea buckthorn seed oil on hyperhomocysteinemia-induced cardiac dysfunction in mice. Simultaneously, H&E and Masson staining were used to assess myocardial structure and fibrosis, and immunohistochemistry was used to assess collagen deposition. The results showed that sea buckthorn seed oil can effectively improve hyperhomocysteinemia-induced cardiac dysfunction and reduce the degree of myocardial fibrosis in mice through both treatment and prevention. Furthermore, transcriptomics analysis, qPCR, and Western blot results indicated that sea buckthorn seed oil exerts its anti-fibrotic effect by inhibiting the expression of fibrosis markers such as α-SMA, type I collagen, and type III collagen in myocardial tissue.

[0021] In this invention, the drug preferably comprises solid dosage forms and / or liquid dosage forms. The solid dosage forms preferably include at least one of the following: tablets, soft capsules, powders, granules, powders, and pellets. The liquid dosage forms preferably include at least one of the following: oral solutions, oral suspensions, oral emulsions, and syrups. This invention does not impose any particular limitation on the preparation method of the drug; any drug well known in the art can be used.

[0022] In this invention, the drug preferably further includes pharmaceutically acceptable excipients. The pharmaceutically acceptable excipients preferably include at least one of the following: diluents, fillers, binders, wetting agents, disintegrants, lubricants, flow aids, flavoring agents, preservatives, and solubilizers. The diluents and / or fillers are mainly used in solid dosage forms (pills, tablets, capsules, granules) to increase the volume of the preparation for easier molding, or to adjust the concentration of the active ingredient. The diluents and fillers include starches, sugars or sugar alcohols, and inorganic substances. The starches include corn starch, potato starch, and wheat starch. The sugars or sugar alcohols include sucrose, lactose, mannitol, and sorbitol. The inorganic substances include microcrystalline cellulose (MCC). The binders or wetting agents are used to bind the traditional Chinese medicine powder or extract into granules, ensuring that the preparation is not easily broken after molding. The wetting agents (which are themselves non-sticky but induce material stickiness) include purified water. The binders include refined honey, beeswax, starch paste, hydroxypropyl methylcellulose (HPMC), povidone (PVP), and sodium carboxymethyl cellulose. The disintegrants include those that promote rapid disintegration of solid dosage forms in the gastrointestinal tract, releasing the drug to improve absorption efficiency; these are commonly used in tablets and capsules. The disintegrants include crospovidone (PVPP), crospovidone sodium carboxymethyl cellulose (CCMC-Na), and sodium carboxymethyl starch (CMS-Na). The lubricants include magnesium stearate, calcium stearate, and stearic acid. The flow aids include talc and / or micronized silica gel. The flavoring agents and aromatizers are used to mask the odor of the active ingredient and improve the palatability of oral preparations; these are commonly used in granules, oral liquids, and chewable tablets. The sweeteners include steviol glycosides, glycyrrhizin, aspartame, and sucrose. The preservatives and solubilizers are commonly used in traditional Chinese medicine liquid preparations (oral liquids, mixtures, syrups) to prevent microbial contamination or promote the dissolution of poorly soluble components. The preservatives include potassium sorbate, sodium benzoate, and parabens. The cosolvents include polyethylene glycol (PEG), Tween 80, and β-cyclodextrin.

[0023] In this invention, the sea buckthorn seed oil content in the drug is preferably 5% or more, and can be 8% to 95%, 10% to 50%, or 15% to 30%.

[0024] The following examples illustrate the application of sea buckthorn seed oil provided by the present invention in the preparation of drugs for the prevention and / or treatment of cardiovascular diseases, but these examples should not be construed as limiting the scope of protection of the present invention.

[0025] Example 1 A method for preparing sea buckthorn seed oil includes the following steps: After the dried sea buckthorn seeds are sieved by gravity and air separation to remove impurities, the shells are separated from the kernels by mechanical means to obtain sea buckthorn seeds, which are then dried at a low temperature of 50℃ for later use. After pressing the sea buckthorn seeds at 55℃ and 27MPa, the crude oil and seed meal are separated by gradient pressure reduction in a separator. The seed meal was subjected to supercritical CO2 extraction with the following parameters: extraction at 30 MPa and 42℃ for 2.5 h to obtain oil fraction; Separate the oil fraction; combine the oil fraction and crude oil at a volume ratio of 3:7, and then perform low-temperature (≤80℃) vacuum degumming, deacidification, decolorization (using a composite adsorbent of activated clay and attapulgite (volume ratio 3:1)) at a vacuum degree ≥0.095 MPa, dewaxing and dehydration treatment under the protection of an inert gas (such as nitrogen), and then filter through a 0.22μm filter membrane to obtain sea buckthorn seed oil.

[0026] Example 2 A method for preparing sea buckthorn seed oil soft capsules 1. Ingredients Contents: 100 g of sea buckthorn seed oil (prepared according to the method of Example 1); Capsule (gelatin) materials: 100 g gelatin, 50 g glycerin, 100 g purified water, 2 g titanium dioxide (light-blocking agent).

[0027] 2. Preparation process a) Preparation of the adhesive solution: Add gelatin, glycerin, and purified water to a dissolving tank, heat to 73°C with stirring until completely melted into a homogeneous and transparent adhesive solution. Add titanium dioxide and continue stirring until evenly dispersed. Then, keep the adhesive solution at 60°C and allow it to stand to remove air bubbles, then set aside for later use.

[0028] b) Content preparation: Sea buckthorn seed oil prepared according to the method of Example 1, without adding any other excipients, is used as the content of the soft capsules.

[0029] c) Capsule Forming: Using a soft capsule capsule forming machine, the insulated gel and sea buckthorn seed oil contents are injected into the equipment separately. Under the control of the mold, the gel forms two continuous rubber sheets, and the sea buckthorn seed oil is quantitatively injected between the two rubber sheets. After rolling and pressing, and cutting, sealed oval soft capsules are formed.

[0030] d) Shaping and drying: Place the shaped soft capsules in a roller and roll them for 4-6 hours at a temperature of 20-25℃ and a relative humidity of <30%. Then transfer them to a drying tunnel and dry them at a temperature of 30-35℃ and a relative humidity of <20% until the moisture content of the capsule shell meets the requirements (usually 12%-16%).

[0031] e) Cleaning and sorting: Clean the surface of the soft capsules with 95% ethanol to remove residual oil stains. After drying, sort the capsules, removing those that are irregularly shaped, leaking oil, or have air bubbles in the shell, to obtain the finished sea buckthorn seed oil soft capsules.

[0032] Sea buckthorn seed oil soft capsules finished product specifications: Each soft capsule is filled with 100mg of pure sea buckthorn seed oil.

[0033] Example 3 Establishment of an animal model of hyperhomocysteinemia (HHcy) and validation of drug efficacy 1. Modeling methods Thirty SPF-grade 5-week-old C57 female mice, weighing 18-20g, were purchased from the Animal Experiment Center of Ningxia Medical University and housed in the university's animal facility under standard laboratory conditions. The experimental procedures were approved by the Ningxia Medical University Experimental Animal Ethics Committee and followed the "Regulations for the Management of Experimental Animals." The mice had free access to food and water, a 12-hour day-night cycle, a temperature of 18-22℃, and a relative humidity of 60%-80%.

[0034] 1.1 Experimental Grouping After one week of acclimatization, the mice were randomly divided into the following groups: blank control group (Control group, n=8) and model group (n=32); the Control group was fed with normal mouse feed throughout the process.

[0035] The modeling groups were divided into: a model control group (Model group, n=8), a sea buckthorn seed oil treatment group (SSO group, n=8), a sea buckthorn seed oil prevention group (Pre-SSO), and a folic acid positive control group (FA group, n=8). The modeling groups were fed a high-methionine diet containing 2% L-methionine for 8 weeks to induce the HHcy model. During the modeling period, the Pre-SSO group mice were administered sea buckthorn seed oil by gavage.

[0036] Eight weeks after modeling, three mice were randomly selected from the Control and Model groups. Blood was collected after anesthesia and serum was separated by centrifugation (3000 r / min, 10 min, 4℃). Serum Hcy concentration was measured using a biochemical Hcy detection kit (Jianglai Biotechnology (catalog number: JT-T1120)). The HHcy and myocardial fibrosis model was considered successfully established when the serum Hcy level in the model group was ≥15 μmol / L (usually ≤15 μmol / L in the Control group), and Masson staining of myocardial tissue showed significantly higher collagen deposition than in the Control group.

[0037] After successful model establishment, the FA group was administered folic acid by gavage, the SSO group was administered sea buckthorn seed oil by gavage, and the model group and control group were administered an equal volume of physiological saline by gavage for four weeks. During model establishment and treatment, indicators such as body weight, random blood glucose, and food intake were monitored. If any mice died or showed abnormal health (such as a sudden drop in body weight >20% or loss of activity), it was recorded in a timely manner and a replacement batch of mice was added.

[0038] Control group: Continued to be fed normal feed, and received an equal volume of physiological saline via intraperitoneal injection daily, and were administered the same medication as the treatment group.

[0039] Model control group: fed with a high methionine diet and administered an equal volume of physiological saline by gavage daily (0.1 mL / 10 g body weight). Folic acid treatment group (FA): fed with high methionine diet and administered folic acid by gavage after successful modeling. The dosage was the clinical conversion dose (converted based on mouse and human surface area) of 0.5 mg / kg body weight.

[0040] Sea buckthorn seed oil treatment group (SSO): After successful modeling, sea buckthorn seed oil was administered by gavage, with the dosage set at 200 mg / kg body weight based on the results of the preliminary experiment. Seabuckthorn seed oil prevention group (Pre-SSO): The diet consisted of feeding with a high methionine diet and gavage administration of seabuckthorn seed oil during modeling. The dosage was set at 200 mg / kg body weight, based on the results of the pre-experiment.

[0041] 1.2 Animal Experiment Detection Indicators 1.2.1 Observe the mice's food intake, weight gain, and random blood glucose levels; 1.2.2 Echocardiography of mice to assess cardiac function; 1.2.3 Biochemical methods were used to detect serum homocysteine ​​(Hcy) levels in mice; 1.2.4 Serum ROS, MDA, and other oxidative stress-related indicators were detected by ELISA. 1.2.4 H&E staining to detect changes in myocardial morphology; 1.2.5 Masson staining and immunohistochemistry were used to observe collagen fiber deposition in mouse myocardium; 1.2.6 Western Blot was used to detect the expression of key proteins and mRNAs of α-SMA, collagen type I, collagen type III fibrosis markers and related pathways.

[0042] 1.3 Transcriptome Sequencing Total RNA was extracted from myocardial samples of each group of mice, and its purity and integrity were verified. Sequencing libraries were constructed through mRNA enrichment, fragmentation, cDNA synthesis, adapter ligation, and PCR amplification. After the libraries met quality standards, high-throughput sequencing was performed using sequencing platforms such as Illumina. The raw data was filtered to obtain clean reads, which were then subjected to reference genome alignment, gene expression quantification, and differentially expressed gene screening (|log2FC|≥1 and...). P <0.05) and GO / KEGG functional enrichment analysis were used to ultimately identify key genes and pathways related to the research objectives.

[0043] 1.4 Statistical Analysis Methods The obtained data were statistically analyzed using Prism 10.0 statistical analysis software. Quantitative data were expressed as mean ± standard deviation. The data is represented by (). Comparisons among multiple sample means were performed using one-way ANOVA for completely randomized design data. Multiple comparisons among multiple sample means were performed using the LSD-t test. Pairwise comparisons between groups were performed using the Steel-Dwass test. The significance level was set at α = 0.05. P <0.05 is statistically significant.

[0044] 2. Results Figure 1 The study demonstrated the successful construction of an HHcy model by feeding the animal with a 2% high-methionine diet for eight weeks. Figure 2 By observing changes in cardiac structure, motion status, and cardiac function indicators, sea buckthorn seed oil treatment can improve myocardial function in Hhcy mice. Sea buckthorn seed oil can effectively improve hyperhomocysteinemia-induced cardiac dysfunction in mice and reduce the degree of myocardial fibrosis.

[0045] Figure 3 The results of serum homocysteine ​​concentration detection in mice of different groups were presented. Compared with the control group or the model group, the Hcy concentration in the prevention group was significantly reduced. At the same time, the Hcy concentration in the folic acid group and the treatment group was significantly reduced compared with the model group. However, the heart weight / body weight ratio, blood glucose level and age-body weight growth curve of mice did not change significantly.

[0046] Figure 4 H&E and Masson staining results showed that sea buckthorn seed oil could improve myocardial structure and fibrosis in HHcy mice. Figure 5 Immunohistochemical results showed that sea buckthorn seed oil could improve collagen deposition in the myocardium of HHcy mice. This indicates that administration of sea buckthorn seed oil can alleviate the degree of HHcy-induced myocardial fibrosis.

[0047] qPCR and Western Blot results showed that the anti-fibrotic effect of sea buckthorn seed oil is related to its inhibition of the expression of fibrotic markers such as α-SMA, type I collagen and type III collagen molecules in myocardial tissue.

[0048] Figure 6 and Figure 7Transcriptome results were analyzed, revealing overlapping genes between the "upregulated genes in the model group vs. control group" and the "downregulated genes in the sea buckthorn treatment group vs. model group." Enriched entries and their scores, along with significantly enriched signaling pathways, were categorized by biological process (BP), cellular component (CC), and molecular function (MF). α-SMA was identified as a differentially expressed gene. Analysis of α-SMA expression levels in different treatment groups showed that, compared to the control group, α-SMA expression was significantly enhanced in the model group. Conversely, compared to the model group, α-SMA expression was significantly reduced after folic acid and sea buckthorn seed oil interventions, suggesting that both interventions could inhibit myocardial fibroblast activation and alleviate myocardial fibrosis.

[0049] The results of the above examples show that sea buckthorn seed oil can be used to develop or prepare functional foods or drugs for the prevention and adjuvant treatment of HHcy-related cardiovascular diseases, providing a new option for the prevention and treatment of HHcy-related cardiovascular diseases.

[0050] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of sea buckthorn seed oil in the preparation of drugs for the prevention and / or treatment of cardiovascular diseases.

2. The application according to claim 1, characterized in that, The cardiovascular diseases mentioned include cardiovascular diseases associated with hyperhomocysteinemia.

3. The application according to claim 2, characterized in that, The cardiovascular diseases associated with hyperhomocysteinemia include myocardial fibrosis induced by hyperhomocysteinemia.

4. The application according to claim 2, characterized in that, Prevention of cardiovascular disease associated with hyperhomocysteinemia includes preventing or delaying the onset of myocardial fibrosis induced by hyperhomocysteinemia.

5. The application according to claim 1, characterized in that, The drug includes drugs that improve at least one of the following in hyperhomocysteinemia: myocardial structure, degree of myocardial fibrosis, and myocardial collagen deposition.

6. The application according to any one of claims 1 to 5, characterized in that, The drug includes solid dosage forms and / or liquid dosage forms.

7. The application according to claim 6, characterized in that, The solid dosage form includes at least one of the following: tablets, soft capsules, powders, granules, powders, and pellets.

8. The application according to claim 6, characterized in that, The liquid formulation includes at least one of the following: oral solution, oral suspension, oral emulsion, and syrup.

9. The application according to claim 6, characterized in that, The drug also includes pharmaceutically acceptable excipients.

10. The application according to claim 6, characterized in that, The drug contains more than 5% sea buckthorn seed oil by mass.

Images