Use of seabuckthorn seed oil in preparation of anti-myocardial fibrosis drugs

Abstract

The application provides application of seabuckthorn seed oil in preparation of a drug for inducing myocardial fibrosis of hyperhomocysteinemia. Existing folic acid and B vitamins can only reduce blood homocysteine, and cannot reverse myocardial collagen deposition and fibrosis damage. The application is proved by double models of rats and mice that seabuckthorn seed oil can improve heart function, relieve ventricular pathological dilation, repair myocardial tissue pathological damage, reduce interstitial collagen accumulation, and inhibit fibroblast activation by down-regulating alpha SMA and type I / III collagen to block the fibrosis process. The raw material of the application is natural and safe, and can be processed into various oral preparations, and is used for prevention and treatment of myocardial fibrosis of hyperhomocysteinemia population, and has good industrialization value.

Description

[0001] This invention relates to the field of novel uses of pharmaceuticals, specifically to the application of sea buckthorn seed oil in the preparation of anti-myocardial fibrosis drugs, and more particularly to the application of sea buckthorn seed oil in the preparation of drugs for myocardial fibrosis caused by hyperhomocysteinemia. Background Technology

[0002] Myocardial fibrosis is a common pathological process in the progression of various cardiovascular diseases to heart failure, characterized by activation of myocardial fibroblasts, excessive deposition of extracellular matrix, and decreased myocardial compliance. Recent studies have found that hyperhomocysteinemia (HHcy) is not only an independent risk factor for atherosclerosis but can also directly induce myocardial interstitial fibrosis, leading to ventricular remodeling and decreased cardiac function. Currently, there are no effective interventions for HHcy-related myocardial damage in clinical practice. Conventional homocysteine-lowering therapies (such as folic acid and B vitamin supplementation) can only reduce homocysteine ​​levels in the blood, and their effect on reversing existing myocardial fibrosis is very limited. There is an urgent need to develop targeted intervention drugs with direct anti-fibrotic activity.

[0003] Sea buckthorn seed oil is rich in various bioactive components, including ω-3 / ω-6 polyunsaturated fatty acids, flavonoid aglycones, tocopherols, and phytosterols, which are known to have antioxidant, anti-inflammatory, and vascular protective effects. However, there is no existing systematic efficacy evaluation or usage record of sea buckthorn seed oil specifically targeting HHcy-induced myocardial fibrosis, and its ability to improve homocysteine-induced myocardial structural disorders and collagen deposition remains inconclusive. Summary of the Invention

[0004] This invention aims to provide a new pharmaceutical use for sea buckthorn seed oil, namely its application in the preparation of drugs for the prevention and / or treatment of myocardial fibrosis (especially hyperhomocysteinemia-related myocardial fibrosis), filling the gap in existing technologies for natural drug interventions against HHcy-induced myocardial fibrosis, and providing a safer and longer-lasting intervention for this type of disease. Technical solution

[0005] To achieve the above objectives, the present invention adopts the following technical solution: Firstly, it provides the application of sea buckthorn seed oil in the preparation of drugs for the prevention, delay, or treatment of myocardial fibrosis.

[0006] Furthermore, the myocardial fibrosis is myocardial fibrosis induced by hyperhomocysteinemia.

[0007] Furthermore, the applications include the preparation of drugs for improving the pathological structure of myocardial tissue, reducing collagen volume fraction, or inhibiting the expression of fibrosis markers.

[0008] Furthermore, the sea buckthorn seed oil inhibits the abnormal activation of myocardial fibroblasts by downregulating the expression levels of α-SMA, type I collagen, and type III collagen in myocardial tissue, thereby blocking the progression of myocardial fibrosis.

[0009] Secondly, it provides the application of sea buckthorn seed oil in the preparation of drugs that improve cardiac remodeling caused by hyperhomocysteinemia, which can enhance cardiac contractile function and reduce the degree of pathological ventricular dilation.

[0010] Thirdly, a pharmaceutical composition is provided comprising an effective amount of sea buckthorn seed oil and a pharmaceutically acceptable carrier, said pharmaceutical composition for the prevention or treatment of myocardial fibrosis associated with hyperhomocysteinemia.

[0011] The pharmaceutical composition can be formulated into oral solid dosage forms (such as soft capsules, tablets, granules, or pellets) or oral liquid dosage forms (such as oral emulsions, suspensions, or syrups).

[0012] The mass percentage of sea buckthorn seed oil in the pharmaceutical composition is 1% to 90%, preferably 5% to 70%, and more preferably 6% to 70%. When the sea buckthorn seed oil content is less than 6%, the active ingredient in a unit dosage form is difficult to reach the onset threshold, requiring an increase in dosage and affecting patient compliance; when the content is higher than 70%, the formability and stability of the formulation decrease significantly, which is not conducive to industrial production. Beneficial effects

[0013] This invention is the first to systematically verify and demonstrate in rat and mouse dual animal models that sea buckthorn seed oil has a significant intervention effect on HHcy-induced myocardial fibrosis, specifically as follows: It can increase left ventricular ejection fraction and fractional shortening, reduce pathological ventricular dilation, and reverse cardiac function damage. It improves the disordered arrangement of cardiomyocytes, reduces abnormal collagen deposition in the myocardial interstitium, and lowers the myocardial collagen volume fraction. It dose-dependently downregulates the expression levels of profibrotic factors such as α-smooth muscle actin (α-SMA) and type I / III collagen, thereby blocking abnormal fibroblast activation pathways.

[0014] This invention broadens the medicinal scope of sea buckthorn seed oil, provides a new intervention strategy for HHcy-related cardiovascular diseases, and sea buckthorn seed oil is a natural edible raw material with no obvious toxic side effects when taken for a long time. It is suitable for the long-term prevention and chronic disease intervention needs of high-risk groups of hyperhomocysteinemia. The raw material is easy to obtain and the preparation of the formulation is easy, which has great potential for industrialization. Attached Figure Description

[0015] Figure 1 The results of M-mode echocardiogram images and cardiac function parameters of mice in each experimental group are presented. Detailed Implementation

[0016] The present invention is further illustrated below by way of examples, but the examples are not intended to limit the present invention.

[0017] Dried sea buckthorn seeds were collected, cleaned, and dried at 60℃ until the moisture content was ≤8%. Cold pressing was performed using a screw press at a pressure controlled at 50–60 MPa to obtain crude oil. The crude oil underwent centrifugal slag removal, degumming, deacidification, and decolorization refining processes to obtain the finished sea buckthorn seed oil. GC-MS analysis showed that the oil contained ≥82% total unsaturated fatty acids, including approximately 36% linoleic acid and approximately 28% α-linolenic acid.

[0018] Formula 1: Take 60 parts by weight of sea buckthorn seed oil, 30 parts by weight of gelatin, 8 parts by weight of glycerin, and an appropriate amount of purified water. Mix the gelatin, glycerin, and purified water and heat to dissolve to prepare the outer shell. Use the sea buckthorn seed oil directly as the contents. Prepare soft capsules using a rotary molding method. Each capsule contains approximately 500mg of net contents and contains no less than 300mg of sea buckthorn seed oil.

[0019] Formula 2: Take 28% sea buckthorn seed oil, 62% refined vegetable oil, 5% beeswax, and 5% vitamin E by weight percentage, mix them thoroughly to obtain the contents, and press them into a gelatin shell made of gelatin, glycerin, and purified water to obtain a stable soft capsule product.

[0020] The following ingredients are added by weight percentage: 12% sea buckthorn seed oil, 5% emulsifier, 2% sweetener, and 0.1% antioxidant. Purified water is added to bring the total volume to 100%. First, the sea buckthorn seed oil and oil phase excipients are mixed and heated to dissolve to prepare the oil phase. Then, the aqueous phase excipients are added to purified water and fully dissolved to prepare the aqueous phase. After the two phases are mixed, they are pre-emulsified by high-speed shearing. Then, they are homogenized three times under a high-pressure homogenizer at a pressure of 20-30 MPa to obtain a uniform and stable oral emulsion product of sea buckthorn seed oil.

[0021] Take 15% sea buckthorn seed oil by weight, and combine it with 50% dextrin, 25% sucrose powder, and 10% hydroxypropyl methylcellulose. Mix the sea buckthorn seed oil and all solid excipients thoroughly and evenly, moisten with an appropriate amount of ethanol, granulate, dry and granulate, and then package to obtain a granule product that can be directly dissolved in water.

[0022] Take 8% sea buckthorn seed oil by mass percentage, and mix it with 0.5% xanthan gum, 1.0% sodium carboxymethyl cellulose, 5% sorbitol, and 0.2% sodium benzoate, then add purified water to make up to 100%. First, add xanthan gum, sodium carboxymethyl cellulose, sorbitol, and sodium benzoate to purified water and stir thoroughly to prepare an aqueous phase. Then, slowly add sea buckthorn seed oil to the aqueous phase, mix by high-speed shearing, and then homogenize under high pressure to obtain a uniformly dispersed oral suspension of sea buckthorn seed oil.

[0023] SPF-grade male SD rats (8 weeks old, weighing 220–250 g) were randomly divided into 4 groups (n=12 per group) after 1 week of acclimatization feeding: Normal control group: administered an equal volume of physiological saline by gavage daily; Model group: A hyperhomocysteinemia model was established by daily gavage administration of L-homocysteine ​​150 mg / kg; Low-dose group of sea buckthorn seed oil: 80 mg / kg of sea buckthorn seed oil was administered by gavage daily during modeling; High-dose group of sea buckthorn seed oil: 320 mg / kg of sea buckthorn seed oil was administered by gavage daily during modeling.

[0024] The animals were administered the medication continuously for 10 weeks. Echocardiographic parameters were measured 24 hours after the last administration. Subsequently, the animals were sacrificed, and heart tissue was collected for pathological staining, immunohistochemistry, and protein quantification.

[0025] Echocardiogram results (see) Figure 1 The left ventricular ejection fraction (LVEF) in the model group decreased from (72.3±3.6)% in the control group to (47.8±4.2)%. P <0.01), the left ventricular fractional shortening (LVFS) decreased from (40.5±2.8)% to (24.1±3.3)%. P <0.01), the left ventricular end-diastolic diameter (LVIDd) and end-systolic diameter (LVIDs) were significantly increased ( P <0.01). The LVEF in the low-dose sea buckthorn seed oil group recovered to (58.6±3.9)% ( P <0.05%, the high-dose group recovered to (66.4±3.7)% ( P <0.01), LVFS synchronization was significantly improved, and the degree of ventricular dilation was significantly reduced.

[0026] Histopathological results: HE staining showed disordered arrangement of cardiomyocytes, significantly widened interstitial spaces, and inflammatory cell infiltration in the model group; Masson staining showed a large amount of blue collagen deposition in the myocardial interstitium of the model group, with collagen volume fraction (CVF) increasing from (2.1±0.5)% in the control group to (16.8±2.5)%. P <0.01). After intervention with sea buckthorn seed oil, the arrangement of cardiomyocytes tended to be more orderly, collagen deposition was significantly reduced, and CVF in the high-dose group decreased to (6.2±1.7)%. P <0.01).

[0027] Immunohistochemical results: The area of ​​α-SMA positive staining in the myocardial tissue of the model group was significantly increased compared with that of the control group. The area of ​​α-SMA positive staining in the low- and high-dose groups of sea buckthorn seed oil decreased in a dose-dependent manner, further confirming the inhibitory effect of sea buckthorn seed oil on fibroblast activation.

[0028] Western blot results: The expression of α-SMA, Collagen I, and Collagen III proteins in the myocardial tissue of the model group was increased by 2.8 times, 3.1 times, and 2.6 times, respectively, compared with the control group. P <0.01). The expression of the above proteins was significantly decreased in the low-dose group of sea buckthorn seed oil ( P <0.05%, the high-dose group showed a greater reduction, approaching the level of the control group ( P <0.01), showing a clear dose-dependent effect.

[0029] Male C57BL / 6J mice (8 weeks old, weighing 20-25g) were used in a repeated grouping and modeling process. The modeling dose was adjusted to L-homocysteine ​​200mg / kg, and the low and high doses of sea buckthorn seed oil were 100mg / kg and 400mg / kg, respectively, for 8 consecutive weeks. The results were completely consistent with the rat model: the model group showed significantly decreased cardiac function, massive collagen deposition, and high expression of fibrosis marker proteins; both dose groups of sea buckthorn seed oil significantly reversed the above-mentioned lesions. P <0.05 or P <0.01).

[0030] The results of cross-species validation showed that the protective effect of sea buckthorn seed oil against high homocysteine-induced myocardial fibrosis is effective and reproducible across species, and the results are stable and reliable. Industrial applicability

[0031] The sea buckthorn seed oil used in this invention is a natural plant product that has been cultivated and processed on a large scale. The raw material source is abundant, the cold-pressing extraction process is mature, and the quality standards are controllable. All preparations made from it are conventional oral dosage forms, with a simple production process that allows for large-scale production without the need for special equipment. Sea buckthorn seed oil itself possesses food-grade safety, with extremely low toxicity and side effects from long-term use. It can be developed into a drug for intervention in myocardial fibrosis in patients with hyperhomocysteinemia, or it can be prepared as a functional food for daily prevention in high-risk groups, with a wide range of applications.

Claims

1. Application of sea buckthorn seed oil in the preparation of drugs for myocardial fibrosis.

2. The application according to claim 1, characterized in that, The myocardial fibrosis was induced by hyperhomocysteinemia.

3. The application according to claim 1, characterized in that, The sea buckthorn seed oil inhibits the abnormal activation of myocardial fibroblasts and blocks the progression of myocardial fibrosis by downregulating the expression levels of α-SMA, type I collagen, and type III collagen in myocardial tissue.

4. The application according to claim 1, characterized in that, The drug is used to improve myocardial tissue pathological damage, reduce myocardial interstitial collagen deposition, and decrease myocardial collagen volume fraction.

5. The application according to claim 1, characterized in that, The drug is used to increase left ventricular ejection fraction and / or left ventricular fractional shortening, improve cardiac remodeling caused by hyperhomocysteinemia, and reduce the degree of pathological ventricular dilation.

6. The application according to any one of claims 1 to 5, characterized in that, The drug is an oral preparation, which is selected from soft capsules, tablets, granules, pellets, oral emulsions, suspensions or syrups.

7. The application according to claim 6, characterized in that, The oral preparation contains 1% to 90% sea buckthorn seed oil by mass, preferably 5% to 70%.

8. The application according to claim 7, characterized in that, The oral preparation contains 6% to 70% sea buckthorn seed oil by mass.

9. The application according to claim 6, characterized in that, The oral formulation further comprises pharmaceutically acceptable excipients, including at least one of fillers, binders, disintegrants, lubricants, emulsifiers, or suspending agents.

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