Application of irisflorentin in preparation of medicine for treating atherosclerosis

Abstract

The invention discloses an application of irisflorentin in preparation of a medicine for treating atherosclerosis. The molecular formula of the irisflorentin is C20H18O8, the molecular weight of the irisflorentin is 386.4, the chemical name of the irisflorentin is 9-mesoxy-7-(3, 4, 5-trimesoxypheyl)-[1, 3] dioxolo [4, 5-g] chromen-8-one, the irisflorentin can inhibit endothelial cell pyroptosis, the preparation medicine of the irisflorentin can be used as an inhibitor of endothelial cell pyroptosis to treat atherosclerosis, and good guiding significance is provided for developing new medicines for preventing and treating atherosclerosis.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to application of irisin in preparing a drug for treating atherosclerosis. Background Art

[0002] Atherosclerosis is a chronic inflammatory disease of the blood vessel wall that involves pathological changes such as endothelial cell damage, foam cell formation, smooth muscle cell migration and proliferation. Atherosclerosis is extremely harmful, and the cardiovascular and cerebrovascular diseases it causes are the leading cause of death in the world. Although the current prevention and treatment strategies of controlling the risk factors for atherosclerosis (smoking, hypertension, diabetes), anti-platelet aggregation, and lowering low-density lipoprotein cholesterol have achieved significant therapeutic effects, the above-mentioned prevention and treatment strategies still fail to reduce the residual risk of cardiovascular events and have many adverse reactions. Therefore, finding new anti-atherosclerotic drugs has become the focus of current research.

[0003] Endothelial cell damage is the initiating link of atherosclerosis. Reducing endothelial cell damage can prevent early atherosclerosis and is a key means of preventing and treating atherosclerosis. Cell pyroptosis is an inflammatory cell death that mainly depends on the activation of NLRP3, ASC, Caspase-1, and GSDMD. The inflammatory factors released by it can cause cell damage. Therefore, inhibiting endothelial cell pyroptosis can reduce endothelial cell damage and thus improve atherosclerosis. Irisin is a small molecule compound derived from the Guizhou Miao medicine Belamcanda chinensis. There are related studies on the inhibition of inflammatory diseases, but there are no studies on its effect on atherosclerosis.

[0004] At present, there is no report on the effect of irisin on the treatment of atherosclerosis. The present invention explores the effect of irisin on atherosclerosis from the perspective of inhibiting endothelial cell pyroptosis, which provides a good guiding significance for the development of new drugs for the prevention and treatment of atherosclerosis. Summary of the invention

[0005] The present invention aims to provide the use of irisin in the preparation of a drug for treating atherosclerosis. From the perspective of inhibiting endothelial cell pyroptosis, the effect of irisin on atherosclerosis is explored.

[0006] The technical solution of the present invention:

[0007] Application of irisin in the preparation of drugs for treating atherosclerosis.

[0008] The molecular formula of the aforementioned irisin is C 20 H 18 O 8, molecular weight is 386.4, chemical name is 9-meth oxy-7-(3,4,5-trimethoxyphenyl)-[1,3]dioxolo[4,5-g]chromen-8-one, structural formula is:

[0009]

[0010] The aforementioned drugs can inhibit endothelial cell pyroptosis.

[0011] The aforementioned drug for treating atherosclerosis is a corresponding drug prepared with irisin as an active ingredient and pharmaceutically acceptable adjuvants.

[0012] The aforementioned drugs for treating atherosclerosis are inhibitors of endothelial cell pyroptosis.

[0013] The aforementioned pharmaceutical preparations include any one or more of granules, powders, capsules, tablets, mixtures or oral solutions.

[0014] A drug for treating atherosclerosis, wherein the drug raw material contains irisin.

[0015] A medicine for treating atherosclerosis, which is mainly prepared from irisin.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention is the first to discover the use of irisin as a drug for treating atherosclerosis. The irisin of the present invention has been experimentally proven to improve atherosclerosis in Apoe- / - mice fed a high-fat diet and inhibit ox-LDL-induced pyroptosis of human umbilical vein endothelial cells, that is, irisin can slow down atherosclerosis by inhibiting endothelial cell pyroptosis. This is a new discovery of the pharmacological effects of irisin, and provides an effective strategy for preparing drugs for treating atherosclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This study aimed to investigate the effects of irisin on aortic atherosclerosis in Apoe- / - mice fed a high-fat diet.

[0019] Figure 2 This is the effect of irisin on the number of pyroptotic cells in human umbilical vein endothelial cells induced by ox-LDL.

[0020] Figure 3 This is the effect of irisin on ox-LDL-induced pyroptosis molecules in human umbilical vein endothelial cells.

[0021] Figure 4This is the effect of irisin on ox-LDL-induced pyroptosis molecule mRNA in human umbilical vein endothelial cells. DETAILED DESCRIPTION

[0022] The present invention will be further described below in conjunction with the embodiments, but they are not intended to limit the present invention.

[0023] Example 1: Take tectorius flavonoids (purchased from the market), add appropriate amounts of conventional auxiliary materials, mix well, dry, sterilize, put into hard capsules, and package to prepare capsules. Each capsule contains 40 mg of tectorius flavonoids.

[0024] Dosage and Administration: Take 1 tablet 3 times a day for the treatment of atherosclerosis.

[0025] Example 2: Take tectorius flavonoids (purchased from the market), add appropriate amounts of conventional auxiliary materials, mix well, make granules, dry, and press into tablets. Each tablet contains 40 mg of tectorius flavonoids.

[0026] Dosage and Administration: Take 1 tablet 3 times a day for the treatment of atherosclerosis.

[0027] Example 3: Take irisin (purchased from the market), add appropriate amount of common auxiliary materials, make granules, dry, and granulate to make granules. Granules 4g / bag, irisin content in the granules is 10mg / g.

[0028] Dosage and Administration: Take 1 bag 3 times a day for the treatment of atherosclerosis.

[0029] Example 4: Take irisin (purchased from the market), add an appropriate amount of solubilizer, grind, add a small amount of water to dilute, mix, add flavoring agent and preservative, mix, add water to a specified amount, filter, mix, package, sterilize, and prepare an oral solution. The content of irisin in the oral solution is 4 mg / ml.

[0030] Dosage and Administration: Take 10 ml three times a day for the treatment of atherosclerosis.

[0031] Example 5: Take irisin (purchased from the market), grind, purify, filter, concentrate, add appropriate flavoring agents and preservatives, package, sterilize, and prepare a mixture. The content of irisin in the mixture is 4 mg / ml.

[0032] Dosage and Administration: Take 10 ml three times a day for the treatment of atherosclerosis.

[0033] Example 6: Take irisin (purchased from the market), crush, sieve, add appropriate amount of common auxiliary materials, mix, divide into doses, package, and prepare powders. The powder is 4g / pack, and the content of irisin in the powder is 10mg / g.

[0034] Dosage and Administration: Take 1 packet 3 times a day for the treatment of atherosclerosis.

[0035] The present invention has done a lot of analysis and verification experiments, and the following are the results of the experimental research of the present invention:

[0036] 1. Effect of irisin on aortic atherosclerosis in Apoe- / - mice fed a high-fat diet

[0037] Six-week-old 18-20g Apoe- / - male mice were divided into three groups: control group (Control), high-fat diet group (HF), high-fat diet + irisin group (HF+IFT), 6 mice in each group. The control group was fed with a normal diet; the high-fat diet group was fed with a high-fat diet (1.3% cholesterol, 16.2% fat, 10.8% sucrose, 9.6% casein, 1.7% premix, 0.3% bile salt, 0.1% methylthiouracil, 2% maltodextrin, 58% basic feed); the high-fat diet + irisin group, on the basis of the high-fat diet, was given irisin 25mg / kg / d by gavage; the feeding period was 12 weeks.

[0038] On the termination day of the experiment, mice were fasted for 12 h and anesthetized with 3% pentobarbital intraperitoneally. They were killed by cervical dislocation and fixed in supine position on the operating table to expose the chest and abdominal cavities. The sternum and ribs were cut open to expose the heart and aorta. Normal saline was slowly perfused into the left ventricle. The end was marked by the whitening of the lungs or the khaki color of the kidneys. Microscissors and microtweezers were used to carefully separate the aorta along the spine to the common iliac arteries on both sides, and the yellow-white adipose tissue around the blood vessels was removed. The heart and aorta were severed at the ascending aorta, and the aorta was fixed and preserved in 4% paraformaldehyde.

[0039] The fixed aorta was removed from 4% paraformaldehyde, washed with saline for 30 minutes × 3 times, immersed in 60% isopropanol for 10 minutes × 3 times, and the aorta was placed in Oil Red O working solution for staining for 120 minutes. After staining, the aorta was removed and placed in 60% isopropanol for decolorization until normal vascular tissue was white and atherosclerotic plaques were bright red or orange. Photos were collected using a digital camera.

[0040] result

[0041] Compared with the high-fat diet group, the aortic atherosclerotic plaque area of ​​Apoe- / - mice fed with a high-fat diet and irisin was significantly reduced ( Figure 1 ).

[0042] 2. Effect of irisin on ox-LDL-induced pyroptosis of human umbilical vein endothelial cells

[0043] Human umbilical vein endothelial cells were divided into control group (Control), irisin group (IFT), ox-LDL induced group (ox-LDL), ox-LDL + irisin group (ox-LDL + IFT). The control group did not receive special treatment, irisin at a concentration of 40umol / L was added to the irisin group, ox-LDL at a concentration of 100ug / mL was added to the ox-LDL induced group, and ox-LDL at a concentration of 100ug / mL and irisin at a concentration of 40umol / L were added to the ox-LDL + irisin group.

[0044] (1) After the cells were grouped and treated, trypsin was used to digest the cells, and the cells were collected into centrifuge tubes. The supernatant was removed by centrifugation, and the cells were resuspended in staining buffer. Annexin-V / PI dye was added to stain for 15 min in the dark. The supernatant was removed by centrifugation, and 200 uL of staining buffer was added to resuspend the cells. The number of pyroptotic cells was detected by flow cytometry.

[0045] (2) After the cells were grouped and treated, they were lysed on ice for 30 min using cell lysis buffer and protease inhibitors, collected in centrifuge tubes, and centrifuged at 4°C and 14,000 rpm. The supernatant was collected and protein loading buffer was added. The expression of pyroptotic molecule proteins was detected using protein immunoblotting.

[0046] (3) After the cells were grouped and treated, total RNA was extracted using an RNA extraction kit, and the total RNA was reverse transcribed into cDNA using a reverse transcription kit. The content of pyroptotic molecule mRNA was detected using a qPCR kit, and GAPDH was used as an internal reference. The relative changes of pyroptotic molecule mRNA were analyzed using the 2-ΔΔCT method.

[0047] result

[0048] Compared with the ox-LDL-induced group, the number of pyroptotic cells in human umbilical vein endothelial cells induced by irisin and ox-LDL was significantly reduced ( Figure 2 ), and the expression of pyroptosis molecule proteins decreased ( Figure 3 ), and the mRNA content of pyroptotic molecules decreased ( Figure 4 ).

Claims

1. Application of irisin in the preparation of drugs for the treatment of atherosclerosis.

2. The use according to claim 1, characterized in that: The molecular formula of irisin is C 20 H 18 O8, molecular weight is 386.4, chemical name is 9-methoxy-7-(3,4,5-trimethoxyphenyl)-[1,3]dio xolo[4,5-g]chromen-8-one, structural formula is:

3. The use according to claim 1, characterized in that: The drug can inhibit endothelial cell pyroptosis.

4. The use according to any one of claims 1 to 3, characterized in that: The drug for treating atherosclerosis is a corresponding drug prepared by using irisin as an active ingredient and pharmaceutically acceptable adjuvants.

5. The use according to claim 4, characterized in that: The drug for treating atherosclerosis is an inhibitor of endothelial cell pyroptosis.

6. The use according to claim 4, characterized in that: The pharmaceutical preparation includes any one or more of granules, powders, capsules, tablets, mixtures or oral solutions.

7. A drug for treating atherosclerosis, characterized in that: The drug raw material contains irisin.

8. A drug for treating atherosclerosis, characterized in that: The medicine is mainly prepared from irisin.

Images