Application of salidroside in preparation of medicine for inhibiting thoracic aortic dissection aneurysm

Abstract

The invention discloses application of salidroside in preparation of a medicine for inhibiting thoracic aortic dissection aneurysm. The invention provides application of salidroside in preparation of a product with any one of the following functions: 1) preventing thoracic aortic dissection aneurysm; 2) the occurrence or progress of thoracic aortic dissection aneurysm is slowed down or inhibited; 3) the occurrence rate of thoracic aortic dissection aneurysm is reduced; and 4) protecting the aorta. The invention finds that the salidroside can significantly improve the occurrence and progress of the thoracic aortic dissection aneurysm of mice, provides a new application approach of the salidroside, and provides a new drug choice for treating the thoracic aortic dissection aneurysm.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to the application of salidroside in the preparation of a medicine for inhibiting thoracic aortic dissecting aneurysm. Background Art

[0002] Thoracic aortic dissection is a high-mortality aortic disease that primarily affects the aortic root, ascending aorta, aortic arch, and descending thoracic aorta. Thoracic aortic dissection refers to a disease in which blood enters the media through a tear in the aortic wall's intima, tearing the aortic wall into two layers and forming a false lumen that can extend along the longitudinal axis of the aorta. Thoracic aortic aneurysm primarily refers to damage to the arterial wall and decreased elasticity, leading to ballooning of the aorta and an increase in internal diameter by more than 50%. It is worth noting that asymptomatic thoracic aortic dissection accounts for 95% of all thoracic aortic dissection cases, and due to the lack of obvious clinical signs, early clinical diagnosis and treatment are particularly important for thoracic aortic dissection.

[0003] Thoracic aortic dissection is a life-threatening disease that requires emergency treatment. Traditional treatment options mainly rely on thoracic and abdominal surgery, but the surgery is traumatic, with high perioperative mortality and complications, and is associated with a high risk of long-term death. In addition, endovascular repair and drug intervention are also common methods for treating thoracic aortic dissection aneurysms. For low-risk patients, drug treatment is preferred, and close follow-up observation is performed. High-risk patients often choose surgery or interventional treatment. Current drug treatment mainly controls heart rate and blood pressure, reduces shear force on the aorta, stabilizes hemodynamics, and reduces the risk of aortic rupture. Therefore, there is a need for safer and more effective drugs to inhibit thoracic aortic dissection aneurysms.

[0004] Salidroside is an important active ingredient in the Tibetan medicine Rhodiola rosea and is also a small molecule natural compound with the chemical formula: tyrosol 8-O-β-D-glucoside (C 14 H 20 O7)( Figure 1 A) Multiple studies have shown that salidroside has multiple benefits, including hypoxia resistance, antioxidant activity, lipid-lowering, blood sugar-lowering, and immune-enhancing effects. However, whether salidroside can inhibit the progression of thoracic aortic dissection remains unclear. Summary of the Invention

[0005] The present invention aims to provide an application of salidroside in the preparation of a medicament for inhibiting thoracic aortic dissecting aneurysm.

[0006] In a first aspect, the present invention provides the use of salidroside in the preparation of a product having any of the following functions:

[0007] 1) Prevent thoracic aortic dissection;

[0008] 2) Slow down or inhibit the occurrence or progression of thoracic aortic dissection;

[0009] 3) Reduce the incidence of thoracic aortic dissection;

[0010] 4) Protect the aorta.

[0011] In the above text, the chemical structure of salidroside is as follows Figure 1 As shown in A.

[0012] The products mentioned above are medicines, compositions, health products, functional foods, foods for special medical purposes or other biological products.

[0013] Among the products mentioned above, the dosage form of the drug is injection, lyophilized powder for injection, aerosol, large infusion, dripping pills, pills, powder, granules, tablets, capsules, oral solution or emulsion.

[0014] In a second aspect, the present invention provides a drug comprising salidroside and any other substance that can serve as a drug carrier;

[0015] The drug has any of the following functions:

[0016] 1) Prevent thoracic aortic dissection;

[0017] 2) Slow down or improve the occurrence or progression of thoracic aortic dissection;

[0018] 3) Reduce the incidence of thoracic aortic dissection;

[0019] 4) Protect the aorta.

[0020] In a third aspect, the present invention provides the use of salidroside and any other substance that can be used as a drug carrier in the preparation of a drug having any of the following functions:

[0021] 1) Prevent thoracic aortic dissection;

[0022] 2) Slow down or improve the occurrence or progression of thoracic aortic dissection;

[0023] 3) Reduce the incidence of thoracic aortic dissection;

[0024] 4) Protect the aorta.

[0025] In the above, the dosage form of the drug is injection, lyophilized powder for injection, aerosol, large infusion, dripping pills, pills, powders, granules, tablets, capsules, oral solution or emulsion.

[0026] The substances that can be used as drug carriers include, but are not limited to, water-soluble carrier materials (such as polyethylene glycol, polyvinyl pyrrolidone, organic acids, etc.), poorly soluble carrier materials (such as ethyl cellulose, cholesterol stearate, etc.), and enteric carrier materials (such as cellulose acetate phthalate and carboxymethyl ethyl cellulose, etc.). Among them, water-soluble carrier materials are preferred. These materials can be used to make a variety of dosage forms, including but not limited to tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal preparations, buccal tablets, suppositories, freeze-dried powder injections, etc. They can be conventional preparations, sustained-release preparations, controlled-release preparations, and various microparticle delivery systems. In order to make unit dosage forms into tablets, various carriers known in the art can be widely used. Examples of carriers include diluents and absorbents, such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate, etc.; wetting agents and binders, such as water, glycerol, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinyl pyrrolidone, etc.; disintegrants. , such as dried starch, alginate, agar powder, brown seaweed starch, sodium bicarbonate with citric acid, calcium carbonate, polyoxyethylene, sorbitan fatty acid esters, sodium lauryl sulfate, methylcellulose, ethylcellulose, etc.; disintegration inhibitors, such as sucrose, tristearin, cocoa butter, hydrogenated oil, etc.; absorption enhancers, such as quaternary ammonium salts, sodium lauryl sulfate, etc.; lubricants, such as talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc. Tablets can also be further prepared as coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer tablets and multilayer tablets. To prepare the unit dosage form into a pill, a wide variety of carriers known in the art can be used. Examples of carriers include diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oils, polyvinylpyrrolidone, gelucine, kaolin, and talc; binders such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste, or flour paste; and disintegrants such as agar powder, dried starch, alginates, sodium lauryl sulfate, methylcellulose, and ethylcellulose. To prepare unit dosage forms as suppositories, a wide variety of carriers known in the art can be used. Examples of carriers include polyethylene glycol, lecithin, cocoa butter, higher alcohols, higher alcohol esters, gelatin, and semi-synthetic glycerides. To prepare unit dosage forms as injectable preparations, such as solutions, emulsions, lyophilized powder injections, and suspensions, all diluents commonly used in the art can be used, including water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters.In addition, to prepare an isotonic injection solution, an appropriate amount of sodium chloride, glucose, or glycerol may be added to the injectable formulation. Conventional solubilizers, buffers, pH adjusters, and the like may also be added. Furthermore, colorants, preservatives, fragrances, flavoring agents, sweeteners, or other materials may be added to the pharmaceutical formulation as needed. The above dosage forms can be administered via injection, including subcutaneous, intravenous, intramuscular, and intracavitary injections; cavitary administration, such as rectal and vaginal; respiratory tract administration, such as nasal; and mucosal administration.

[0027] The subject of the above-mentioned drug is a human or an animal. The animal can be a mammal, including a primate (such as a monkey or chimpanzee), a non-primate (such as a cow, pig, camel, llama, horse, goat, rabbit, sheep, hamster, guinea pig, cat, dog, rat, mouse, horse or whale), or a bird (such as a duck or goose). In one embodiment, the subject is a mouse.

[0028] Beneficial effects of the present invention: The present invention found that salidroside can significantly improve the occurrence and progression of thoracic aortic dissection aneurysms in mice, providing a new application approach for salidroside and a new drug option for the treatment of thoracic aortic dissection aneurysms. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The figure shows that salidroside significantly reduces the occurrence and rupture of thoracic aortic dissection aneurysms induced by BAPN in mice. 1A is the chemical formula of salidroside, 1B is the statistical results of the survival rates of mice in the salidroside treatment group and the control group, 1C is the incidence of thoracic aortic aneurysms / dissections; 1D is the gross photograph of the thoracic aorta of mice in the salidroside treatment group and the control group; 1E is the ultrasound examination results of the thoracic aorta of mice in the salidroside treatment group and the control group.

[0030] Figure 2 Salidroside significantly improved the pathological changes of BAPN-induced thoracic aortic dissection in mice. DETAILED DESCRIPTION

[0031] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0032] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0033] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. The embodiments are used to describe the present invention but not to limit the present invention.

[0034] Experimental animals: 3-week-old male wild-type mice (C57BL / 6J), weighing about 10 g, were purchased from Saiye Biotechnology Co., Ltd. The animals were raised and bred in an SPF-grade animal room.

[0035] Main reagents: Salidroside (Sigma, USA), 3-aminopropionitrile fumarate (BAPN, Sigma, USA).

[0036] Main experimental instruments: Vevo ultra-high resolution multimodal small animal ultrasound photoacoustic imaging system (Fuji, Japan), paraffin slicer (LEICA, Germany), small animal electronic weighing scale.

[0037] Example 1: Application of Salidroside in Inhibiting the Progression of Thoracic Aortic Dissecting Aneurysm

[0038] The inventors screened traditional Tibetan medicine ingredients or small molecule compounds from common plant sources using animal models and found that salidroside can be used as a new drug to inhibit the development of thoracic aortic dissecting aneurysm.

[0039] 1. Mouse Thoracic Aortic Dissection Model and Administration of Salidroside

[0040] The mouse thoracic aortic dissection aneurysm model was established using the BAPN water feeding method as follows:

[0041] Three-week-old wild-type male mice were fed a normal diet and treated with BAPN dissolved in drinking water at a dose of 1 g / kg / day for 4 weeks to induce thoracic aortic dissection.

[0042] Three-week-old wild-type males were randomly divided into two groups:

[0043] One group was the salidroside treatment group (denoted as BAPN+salidroside in the figure): starting from the start of water-feeding with BAPN (denoted as day 0), salidroside (dissolved in normal saline) was intraperitoneally injected at a dose of 100 mg / kg three times a week for 4 weeks.

[0044] The other group was the control group (denoted as BAPN+Vehicle in the figure): the treatment frequency was the same as that of the salidroside treatment group, except that the same volume of normal saline was injected via intraperitoneal injection (Vehicle).

[0045] 2. Observation indicators after administration

[0046] 1. Count the number of mice that survived for 4 weeks in the two groups mentioned above and calculate the survival rate.

[0047] Death of mice during modeling: record the time of death and cause of death (whether death was due to dissection / aneurysm rupture).

[0048] 2. Ultrasonic testing

[0049] Surviving mice from the two groups were examined four weeks later. Mice were anesthetized with 2% isoflurane inhalation and secured in a supine position on a heated ultrasound table. Their chest and abdomen were depilated with depilatory cream and then coated with ultrasound coupling gel. Aortic arch ultrasound was performed using a small animal ultrasound system and a 30 MHz high-frequency probe to assess for enlargement or dissection. A 50% or greater dilation of the thoracic aortic diameter was considered a thoracic aortic aneurysm / dissection.

[0050] Statistics of the incidence of thoracic aortic aneurysm / dissection: The incidence of thoracic aortic aneurysm / dissection is the percentage of the sum of the number of mice that died due to dissection / aneurysm rupture + the number of mice with thoracic aortic aneurysm / dissection confirmed by ultrasound detection to the total number of experimental mice.

[0051] The results showed that the survival rate of mice in the salidroside treatment group was significantly improved ( Figure 1 B); The incidence of thoracic aortic aneurysm / dissection in mice treated with salidroside was 33.3%, which was much lower than 76.7% in the control group, indicating that salidroside inhibited the occurrence of thoracic aortic aneurysm / dissection in mice ( Figure 1 C). Take a general photo ( Figure 1 D) and small animal ultrasound ( Figure 1 E) shows that the aortic lumen of mice in the salidroside-treated group was basically normal.

[0052] 3. Preparation of paraffin sections

[0053] At the end of the ultrasound experiment, mice in the salidroside-treated and control groups were anesthetized with sodium pentobarbital (100 mg / kg) and sacrificed. Cardiac perfusion was performed with heparinized saline to remove residual blood. The aortic arch was excised under a stereomicroscope and fixed in 10% formaldehyde for at least 24 hours. The vessels were removed, dehydrated, and the tissue embedded in paraffin. Serial sections were used for sectioning, with a thickness of 5 μm.

[0054] A. Observation of vascular structure by HE staining

[0055] (1) Paraffin sections are routinely dewaxed with xylene, and then dewaxed with ethanol at different concentrations until the paraffin sections are water-soluble.

[0056] Toluene I (20 min) → Xylene II (20 min) → 100% alcohol (5 min) → 95% alcohol (5 min) → 80% alcohol (5 min) → rinse with tap water to remove alcohol

[0057] (2) Hematoxylin staining for 3-5 minutes, rinse with tap water 3 times

[0058] (3) Hydrochloric acid and ethanol differentiation for 2 seconds, tap water at a low flow rate to return to blue (5 minutes)

[0059] (4) Stain with eosin for 2-3 minutes, rinse with tap water 3 times

[0060] (5) Conventional dehydration, transparency, and sealing: 80% alcohol (5 min) → 95% alcohol (5 min) → 100% alcohol (5 min) → 100% alcohol (5 min) → xylene I (5 min) → xylene II (5 min) → neutral gum sealing

[0061] B. Observe the destruction of elastic fibers in the vascular wall by EVG staining

[0062] (1) Paraffin sections are routinely dewaxed with xylene, and then dewaxed with ethanol at different concentrations until the paraffin sections are water-soluble.

[0063] Toluene I (20 min) → Xylene II (20 min) → 100% alcohol (5 min) → 95% alcohol (5 min) → 80% alcohol (5 min) → rinse with tap water to remove alcohol

[0064] (2) Dye in a greenhouse with the prepared Verhoeff's dye solution for 1-3 minutes until the color becomes dark black, and rinse with tap water

[0065] (3) 2% ferric chloride solution was used for 10-20 seconds. Microscopic observation showed that the elastic fibers were black and the background was gray. Rinse with tap water.

[0066] (4) Van Gieson's solution counterstaining for 10-15 seconds, followed by differentiation with anhydrous ethanol

[0067] (5) Dehydrate with anhydrous ethanol, make transparent with xylene, and seal with neutral gum

[0068] C. Observe the collagen deposition in the vascular wall by Masson staining

[0069] (1) Paraffin sections are routinely dewaxed with xylene, and then dewaxed with ethanol at different concentrations until the paraffin sections are water-soluble.

[0070] Toluene I (20 min) → Xylene II (20 min) → 100% alcohol (5 min) → 95% alcohol (5 min) → 80% alcohol (5 min) → rinse with tap water to remove alcohol

[0071] (2) Hematoxylin staining of cell nuclei: Stain with hematoxylin in Masson staining kit for 5 minutes, wash with tap water, differentiate with 1% hydrochloric acid ethanol for a few seconds, rinse with tap water, and rinse with running water for a few minutes to return to blue.

[0072] (3) Ponceau staining: Stain with Ponceau acid fuchsin solution in the Masson staining kit for 5-10 minutes, then rinse with tap water.

[0073] (4) Rapidly rinse with distilled water. Treat with the phosphomolybdic acid aqueous solution in the Masson staining kit for about 3-5 minutes.

[0074] (5) Aniline blue staining: No need to wash with water, directly use the aniline blue solution in the Masson staining kit to counterstain for 5 minutes

[0075] (6) Differentiation: 1% glacial acetic acid treatment for 1 minute

[0076] (7) Dehydrate with anhydrous ethanol, make transparent with xylene, and seal with neutral gum

[0077] The vascular staining of the mouse model is shown in Figure 2 It can be seen that the arterial lumen of the mice in the salidroside treatment group was smooth and of normal diameter, while the arterial vessels of the mice in the control group (BAPN induced) were significantly dilated, and at the same time, part of the vascular inner wall ruptured, with obvious dissection.

[0078] The above results show that salidroside can reduce the rupture and formation of thoracic aortic dissection aneurysm induced by BAPN in mice, reduce the incidence of thoracic aortic aneurysm / dissection, and prevent thoracic aortic dissection aneurysm, indicating that salidroside has a protective effect on the aorta.

[0079] Finally, it should be noted that the above embodiments are only used to help those skilled in the art understand the essence of the technical solution of the present invention and are not intended to limit the scope of protection of the present invention.

Claims

1. Application of salidroside in the preparation of products having any of the following functions: 1) Prevent thoracic aortic dissection; 2) Slow down or inhibit the occurrence or progression of thoracic aortic dissection; 3) Reduce the incidence of thoracic aortic dissection; 4) Protect the aorta.

2. The product according to claim 1, characterized in that: The product is a medicine, composition, health product, functional food, food for special medical purposes or other biological product.

3. The product according to claim 2, characterized in that: The dosage form of the drug is injection, lyophilized powder for injection, aerosol, large infusion, dripping pills, pills, powders, granules, tablets, capsules, oral solution or emulsion.

4. A drug comprising salidroside and any other substance that can serve as a drug carrier; The drug has any of the following functions: 1) Prevent thoracic aortic dissection; 2) Slow down or improve the occurrence or progression of thoracic aortic dissection; 3) Reduce the incidence of thoracic aortic dissection; 4) Protect the aorta.

5. Use of salidroside and any other substance that can be used as a drug carrier in the preparation of drugs having any of the following functions: 1) Prevent thoracic aortic dissection; 2) Slow down or improve the occurrence or progression of thoracic aortic dissection; 3) Reduce the incidence of thoracic aortic dissection; 4) Protect the aorta.

6. The medicine according to claim 4 or the use according to claim 5, characterized in that: The dosage form of the drug is injection, lyophilized powder for injection, aerosol, large infusion, dripping pills, pills, powders, granules, tablets, capsules, oral solution or emulsion.

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