Application of burdock root polysaccharide in preparation of medicine for treating pulmonary arterial hypertension

Abstract

The invention discloses application of burdock root polysaccharide in preparation of a medicine for treating pulmonary arterial hypertension, and belongs to the technical field of medicines. The burdock root polysaccharide is used as a unique active ingredient to prepare the medicine for treating pulmonary arterial hypertension, the medicine comprises an effective amount of burdock root polysaccharide and pharmaceutically acceptable auxiliary materials, and experimental results show that the burdock root polysaccharide remarkably reduces increase of rat pulmonary arterial pressure induced by monocrotaline, remarkably inhibits increase of right ventricular hypertrophy index, and has a significant effect on treating pulmonary arterial hypertension. The traditional Chinese medicine composition can be used for remarkably relieving pulmonary artery endothelial injury, middle-layer thickening, middle-layer cell hyperproliferation and inflammatory cell infiltration of a pulmonary artery hypertension rat, remarkably relieving the pulmonary artery blood vessel muscle degree of the pulmonary artery hypertension rat, and has the effects of relieving pulmonary artery blood vessel remodeling and reducing pulmonary artery pressure; the compound has a remarkable treatment effect on pulmonary arterial hypertension and right ventricular hypertrophy of rats, and can be applied to preparation of drugs for treating pulmonary arterial hypertension.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to application of burdock root polysaccharide in preparing medicine for treating pulmonary hypertension. Background Art

[0002] Pulmonary hypertension (PH) is a complex progressive cardiopulmonary disease. The exact pathogenesis is still unclear, the patient's survival period is limited, and there is no effective cure in clinical practice. PH is also considered the "cancer" of the cardiovascular field. About 1% of the population worldwide suffers from PH, and the incidence rate increases with age, especially in people over 65 years old, which rises significantly to 10%, and about 50% of patients also have heart failure. As my country gradually enters an aging society, PH has become one of the serious problems affecting residents' health. At present, the treatment of PH mainly focuses on vasodilation, and there is a lack of ideal treatment methods.

[0003] Burdock (Arctium Lappa L.) is a biennial herbaceous plant of the Asteraceae family and a common plant with medicinal and edible properties. Polysaccharide from Arctium lappa.L.root (ALR-PS) is extracted from its root. Studies have shown that burdock root polysaccharide has multiple pharmacological activities such as anti-tumor, anti-allergy, and hypoglycemic. However, no relevant research on the therapeutic effect of burdock root polysaccharide on pulmonary arterial hypertension has been reported so far. Summary of the invention

[0004] In order to solve the problems existing in the prior art, the purpose of the present invention is to provide the use of burdock root polysaccharide in the preparation of a drug for the treatment of pulmonary hypertension. Burdock root polysaccharide has the effect of alleviating pulmonary artery vascular remodeling and lowering pulmonary artery pressure, has a significant therapeutic effect on pulmonary arterial hypertension and right ventricular hypertrophy in rats, and can be used to prepare a drug for the treatment of pulmonary hypertension.

[0005] The object of the present invention is to achieve the following:

[0006] The invention provides application of burdock root polysaccharide in preparing medicine for treating pulmonary hypertension.

[0007] Based on the above technical solution, further, the burdock root polysaccharide is the only active ingredient.

[0008] Based on the above technical solution, further, the medicine comprises an effective amount of burdock root polysaccharide and pharmaceutically acceptable excipients.

[0009] Based on the above technical solution, further, the pharmaceutically acceptable excipients include fillers, diluents, binders, disintegrants, emulsifiers and drug carriers without toxic side effects.

[0010] Based on the above technical solution, further, the dosage form of the drug includes tablets, granules, oral liquid preparations, drops, injection preparations and capsule preparations.

[0011] Based on the above technical solution, further, the drug is prepared in the form of a single-dose drug.

[0012] Based on the above technical solution, further, the single dose of medicine contains 100 to 30,000 mg of burdock root polysaccharide.

[0013] Based on the above technical solution, further, the drug can significantly reduce the increase of pulmonary artery pressure, significantly inhibit the increase of right ventricular hypertrophy index, and significantly alleviate pulmonary artery vascular remodeling.

[0014] The beneficial effects of the present invention compared with the prior art are as follows:

[0015] The burdock root polysaccharide of the invention has low extraction cost and high safety. Experiments have confirmed that the burdock root polysaccharide significantly reduces the increase of pulmonary artery pressure in rats induced by monocrotaline, significantly inhibits the increase of right ventricular hypertrophy index, significantly alleviates pulmonary artery endothelial damage, middle layer thickening, middle layer cell hyperproliferation and inflammatory cell infiltration in rats with pulmonary hypertension, significantly alleviates the degree of pulmonary artery vascular muscularization in rats with pulmonary hypertension, has the effects of alleviating pulmonary artery vascular remodeling and reducing pulmonary artery pressure, and has a significant therapeutic effect on rat pulmonary hypertension and right ventricular hypertrophy, and can be used to prepare a drug for treating pulmonary hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention, the drawings related to the embodiments are briefly introduced below.

[0017] Figure 1 ALR-PS can significantly reduce the increase in pulmonary artery pressure in rats with pulmonary hypertension induced by monocrotaline (MCT), *p<0.05, ***p<0.001.

[0018] Figure 2 ALR-PS can significantly reduce right ventricular hypertrophy in rats with pulmonary hypertension induced by monocrotaline (MCT), *p<0.05, ***p<0.001.

[0019] Figure 3The results show that ALR-PS can significantly alleviate pulmonary artery vascular remodeling in rats with pulmonary hypertension induced by monocrotaline (MCT), where A is HE staining and αSMA immunohistochemistry of rat pulmonary artery (20×, scale: 50μm), B is the media thickness percentage WT%, C is the media area percentage WA%, and D is the relative optical density value of αSMA, *p<0.05, **p<0.01, ***p<0.001.

[0020] Figure 4 The results of pulmonary artery muscularization in rats in each group, where A is the immunohistochemical staining of αSMA (4×, scale: 200 μm), B is the percentage of muscularization, *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0021] The present invention is described in detail below with reference to the embodiments, but the embodiments of the present invention are not limited thereto. Obviously, the embodiments described below are only partial embodiments of the present invention. For those skilled in the art, other similar embodiments obtained without creative labor all fall within the protection scope of the present invention.

[0022] Example 1

[0023] This embodiment provides a method for preparing burdock root polysaccharide (ALR-PS), comprising the following steps:

[0024] (1) Peel and clean the burdock root, slice it, dry it in an oven at 60°C, and grind it into powder.

[0025] (2) Add water at a solid-liquid ratio of 1:15, boil at 80°C for 3 h, filter with gauze and filter paper in turn, collect the filtrate, continue to add water and boil the residue, and repeat this step 3-4 times.

[0026] (3) Collect the filtrate and filter it with ordinary filter paper to initially remove insoluble impurities.

[0027] (4) The filtrate was concentrated by rotary evaporation to an appropriate volume, and 3 volumes of anhydrous ethanol were added while stirring. The mixture was allowed to stand overnight to separate the layers.

[0028] (5) Collect the lower layer of precipitate, dry it by rotary evaporation, and then add appropriate amount of water to fully dissolve the burdock root polysaccharide.

[0029] (6) Repeat steps (4) to (5) 3-4 times, collect the burdock root polysaccharide aqueous solution, and freeze-dry to obtain burdock root polysaccharide.

[0030] (7) After dissolving the dried burdock root polysaccharide in water, the mixture was filtered using a 0.45 μm microporous filter membrane.

[0031] (8) Collect the filtrate, freeze-dry it, and obtain burdock root polysaccharide powder.

[0032] Example 2

[0033] This example investigates the therapeutic effect of the burdock root polysaccharide prepared in Example 1 on rats with pulmonary hypertension induced by monocrotaline.

[0034] (1) Establishment of rat pulmonary hypertension model: A single subcutaneous injection of 60 mg / kg monocrotaline (MCT, C101556-1g, Shanghai Aladdin Biochemical Technology Co., Ltd.) was given to rats.

[0035] (2) Animal Experiment Grouping

[0036] Male adult SD rats (6-8 weeks old, weighing 180-200 g) were purchased from Liaoning Changsheng Biotechnology Co., Ltd. (certificate number: SCXK (Liao) 2020-0001) and randomly divided into the following groups, 8 rats in each group.

[0037] a. Normal group: Rats were injected subcutaneously with 10 ml / kg of normal saline once in the neck, weighed and gavaged with ddH2O (10 ml / kg) every day for 28 consecutive days.

[0038] b. Normal administration group: rats were subcutaneously injected with 10 ml / kg of normal saline in the neck once, weighed and intragastrically administered with ALR-PS (600 mg / kg) once a day for 28 consecutive days.

[0039] c. MCT group: Rats were given a single subcutaneous injection of monocrotaline 60 mg / kg in the neck, weighed and intragastrically administered with ddH2O once a day for 28 consecutive days.

[0040] d. MCT+ALR-PS (300 mg / kg) group: Rats were subcutaneously injected with monocrotaline 60 mg / kg in the neck once, weighed and intragastrically administered with ALR-PS (300 mg / kg) once a day for 28 consecutive days.

[0041] e. MCT+ALR-PS (600 mg / kg) group: Rats were subcutaneously injected with monocrotaline 60 mg / kg in the neck once, and were weighed and intragastrically administered with ALR-PS (600 mg / kg) once a day for 28 consecutive days.

[0042] (3) Detection of right ventricular systolic pressure (RVSP) in rats

[0043] After 28 days of treatment, rats were anesthetized by isoflurane inhalation anesthesia and fixed in supine position. The skin around the neck was prepared. A small incision parallel to the long axis of the neck was made above the right external jugular vein on the right neck of the rat using a surgical scalpel, and the right external jugular vein was bluntly separated. The distal end of the rat external jugular vein was ligated with sterile surgical sutures. A small "V"-shaped incision was made at the proximal end using corneal scissors under a stereomicroscope, and a polyethylene catheter filled with heparin solution was inserted. Under the guidance of the pressure waveform change, the catheter was inserted through the right external jugular vein, right atrium to the right ventricle. The other end of the catheter was connected to a pressure sensor, and the right ventricular systolic pressure (RVSP) of the rat was measured using a BL-420N biofunctional experimental system to evaluate the pulmonary artery pressure level.

[0044] (4) Right ventricular hypertrophy index (Fulton Index) detection

[0045] The rat heart was removed, and the large blood vessels, right atrium, and left atrium connected to the heart were removed using ophthalmic scissors. The right ventricle (RV) and left ventricle + ventricular septum (LV + S) were separated. The excess saline was absorbed with filter paper and then the components were weighed. The Fulton index, the ratio of the right ventricular weight to the left ventricle plus the ventricular septum weight [RV / (LV + S)], was calculated to represent the degree of right ventricular hypertrophy.

[0046] (5) Morphological examination of pulmonary arterial vascular remodeling

[0047] The chest was opened along the ribs on both sides, and pre-cooled saline was injected at a constant rate through the right ventricle using an infusion set. The right auricle was cut open for perfusion. A 10 mL syringe was used to inject 4% paraformaldehyde into the right lung through the right bronchus. The injection volume was appropriate to fill the right lung to a slightly expanded state. The right lung tissue was separated and infiltrated in 4% paraformaldehyde fixative and stored at 4°C for 72 h. It was then paraffin-embedded and sliced ​​at 4 μm thickness. HE staining (C0105S, Bio-Tech Biotech Co., Ltd.) was performed to evaluate morphological changes. The outer diameter, inner diameter, media area, and total vascular area of ​​the pulmonary arterioles were measured using the Image Pro Plus pathological image analysis system, and the media thickness percentage WT% = (outer diameter - inner diameter) / (outer diameter) × 100; area percentage WA% = (media area) / (total vascular area) × 100, representing the degree of pulmonary artery thickening.

[0048] α-actin (αSMA) is a marker protein of smooth muscle cells and a vascular muscularization factor. The sections were subjected to αSMA immunohistochemistry (ab7817, abcam), and the optical density value (relative OD) was used to evaluate the expression of αSMA in the pulmonary artery of each group of rats to determine the proliferation of smooth muscle cells in the middle layer of the pulmonary artery; according to the αSMA-positive expression area of ​​the pulmonary artery, the muscularization ratio of the pulmonary artery with a vessel diameter of <200μm was calculated to evaluate the degree of pulmonary artery muscularization.

[0049] (6) Results

[0050] Figure 1 is the average right ventricular pressure (RVSP) result of each group of rats. The results showed that the right ventricular systolic pressure of rats in the model group induced by monocrotaline was significantly increased (p<0.001). 300mg / kg and 600mg / kg burdock root polysaccharides can effectively alleviate the increase of right ventricular systolic pressure RVSP in rats (p<0.05), and the 600mg / kg dose group was significantly better than 300mg / kg (p<0.05), indicating that burdock root polysaccharide can significantly reduce the increase of pulmonary artery pressure in rats induced by monocrotaline.

[0051] Figure 2 The results of right ventricular hypertrophy index (Fulton Index) of rats in each group showed that the right ventricular hypertrophy index of rats in the model group increased significantly (p<0.001), and the right ventricular hypertrophy index decreased after administration of burdock root polysaccharide. Among them, 600 mg / kg burdock root polysaccharide could significantly inhibit the increase of right ventricular hypertrophy index (p<0.05).

[0052] Figure 3 The lung tissue morphology results of rats in each group. Figure 3 HE staining results of A showed that the rats in the pulmonary hypertension model group had pulmonary artery endothelial cell damage, pulmonary artery lumen stenosis and occlusion, accompanied by a large number of inflammatory cell infiltration, while the lung tissue morphology of rats in the burdock root polysaccharide group was significantly improved. Burdock root polysaccharide can significantly protect vascular endothelial cells, inhibit arterial lumen stenosis, and reduce inflammatory cells. Figure 3 B. Figure 3 C The results showed that the media thickness percentage WT% and media area percentage WA% of the rats in the model group were significantly increased (p<0.001), and the WT% and WA% indexes of the burdock root polysaccharide group were decreased (p<0.05), indicating that burdock root polysaccharide can significantly alleviate the thickening of the pulmonary artery media in rats with pulmonary hypertension.

[0053] The results of αSMA immunohistochemistry in rat pulmonary artery are shown in Figure 3 A. Figure 3As shown in D, the optical density of αSMA in the model group rats was significantly increased (p<0.001), and the optical density of αSMA decreased after administration of burdock root polysaccharide (p<0.05). The results showed that burdock root polysaccharide can significantly alleviate the excessive proliferation of pulmonary artery media cells in rats with pulmonary hypertension.

[0054] Figure 4 The results of pulmonary artery muscularization in rats in each group. After immunohistochemical staining of αSMA in the rat pulmonary artery, the ratio of muscularized vessels with a vessel diameter of <200μm was counted. The results showed that the ratio of muscularized vessels in the model group was significantly increased (p<0.001), and the ratio of muscularized vessels decreased after administration of burdock root polysaccharide. The results showed that burdock root polysaccharide can significantly alleviate the degree of pulmonary artery muscularization in rats with pulmonary hypertension.

[0055] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of burdock root polysaccharide in the preparation of medicines for treating pulmonary hypertension.

2. The use according to claim 1, characterized in that: The burdock root polysaccharide is the only active ingredient.

3. The use according to claim 1, characterized in that: The medicine comprises an effective amount of burdock root polysaccharide and pharmaceutically acceptable auxiliary materials.

4. The use according to claim 3, characterized in that: The pharmaceutically acceptable excipients include fillers, diluents, binders, disintegrants, emulsifiers and drug carriers without toxic or side effects.

5. The use according to claim 1, characterized in that: The dosage forms of the medicine include tablets, granules, oral liquid preparations, drops, injection preparations and capsule preparations.

6. The use according to claim 1, characterized in that: The medicine is prepared in the form of a single-dose medicine, and the single-dose medicine contains 100 to 30,000 mg of burdock root polysaccharide.

7. The use according to claim 1, characterized in that: The drug can significantly reduce the increase of pulmonary artery pressure, significantly inhibit the increase of right ventricular hypertrophy index, and significantly alleviate pulmonary artery vascular remodeling.

Images