CN107308141A

CN107308141A - Application of the dendrophnol and combinations thereof in prevention and treatment atherosclerosis medicine is prepared

Info

Publication number: CN107308141A
Application number: CN201710396048.2A
Authority: CN
Inventors: 王伟; 王倩倩; 吴燕; 王儒情; 徐忠东
Original assignee: Hefei Normal University
Current assignee: Hefei Normal University
Priority date: 2017-05-31
Filing date: 2017-05-31
Publication date: 2017-11-03
Anticipated expiration: 2037-05-31
Also published as: CN107308141B

Abstract

The invention belongs to the new medical use of the active ingredient dendrophnol of Chinese medicine, application of the dendrophnol in prevention and treatment atherosclerosis medicine is prepared is concretely related to.Animal test results show：Dendrophnol has obvious prevention and improvement result for Atherosclerosis Model mouse aorta atherosclerosis plaque forming.Relevant auxiliary materials are equipped with using dendrophnol, health products or medicine that specific aim prevents and treats atherosclerosis are can be made into conventional formulation method.

Description

Dendrophnol and combinations thereof is in prevention and treatment atherosclerosis medicine is prepared Using

Technical field

The invention belongs to the new medical use of the active ingredient dendrophnol of Chinese medicine, dendrophnol is concretely related in system Application in standby prevention and treatment atherosclerosis medicine.

Background technology

Atherosclerosis (Atherosclerosis) property angiocardiopathy be the current cause of the death most common in the world it One.In China, the death toll of current angiocardiopathy induction accounts for the 1/3 of whole death tolls, and its incidence and death Rate is increasing year by year.Pathogenesis of Atherosclerosis is complicated, and inducement is numerous, and its formation is a complicated process.Epidemic disease Data is pointed out, and serum LDL cholesterol level and atherosclerosis are proportionate, and high density fat Cholesterol in albumen is inversely to transport cholesterol to liver processing, it is possible to decrease body cholesterol levels, so as to resist artery Atherosis.It is more complete that current main flow academic viewpoint thinks that inflammation reaction theory is explained the pathomechanism of atherosclerosis Face.Inflammation reaction theory thinks that inflammatory is thin from vascular endothelial cell damage to during the entire process of clinical cardiovascular events occur Intracellular cytokine, such as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemotactic egg - 1 (MCP-1) etc. in vain, the inflammatory reaction of participation is all played an important role.Therefore, regulation blood lipid metabolism and suppression vascular inflammation Reaction provides important thinking for exploitation treatment of atherosclerosis medicine.

At present, 3-Hydroxy-3-methylglutaryl CoA A is mainly used to the clinical treatment of atherosclerosis both at home and abroad (HMG-CoA) reductase inhibitor (Statins), but muscle poison (myopathy, rhabdomyolysis) and the liver enzyme change of its induction Its clinical practice is further limit Deng adverse reaction.Therefore, find and develop safely and effectively Antiatherosclerosis medicine Just it is particularly important.

A kind of dendrophnol (Gigantol), bibenzyl phenolic substance, is extracted from the root of the stem of noble dendrobium, is the medicinal effective of the stem of noble dendrobium One of composition.Have now been found that some evidences illustrate the stem of noble dendrobium and have various biological activity, including enhancing immunity of organisms, side Help the effect such as gastric digestion, anti-cataract, regulation blood glucose, antitumor.By being found to dendrophnol anti-cataract study mechanism, stone Dry measure used in former times phenol can reach the effect of anti-cataract by suppressing the activity of nitric oxide synthase type and its specific binding site Really.

So far, there is not yet using dendrophnol as the relevant report of prevention and treatment atherosclerosis medicine.

The content of the invention

Technical problem：Application of the dendrophnol in prevention and treatment atherosclerosis medicine is prepared is provided.

Technical scheme：Dendrophnol and combinations thereof applying at this in prevention and treatment atherosclerosis medicine is prepared Within the protection domain of invention.

Dendrophnol and combinations thereof applying in protection scope of the present invention in prevention of arterial anti-atherosclerotic agent is prepared Within.

Dendrophnol and combinations thereof applying in protection scope of the present invention in treatment atherosclerosis medicine is prepared Within.

Dendrophnol composition is that oral liquid, injection, tablet, pill, dispersant, capsule is made with the auxiliary material of corresponding formulation Agent, soft capsule, dripping pill, granule, solution, suspending agent, emulsion or microcapsules.Can also be athero- using dendrophnol as prevention of arterial The health products of hardening.

The technical method that the present invention is provided can be dendrophnol single component or be substantially effective containing dendrophnol The medicine of composition and pharmaceutically acceptable adjuvant composition.

Beneficial effect：

The invention provides the dendrophnol to atherosclerosis with preventive and therapeutic action prepare medicament (health products or Medicine), for preventing and treating atherosclerosis.The present invention has progressive and advantage as is evident below：

1st, definite ingredients (dendrophnol)；2nd, with clearly defined objective (atherosclerosis)；3rd, adjustment effect clearly (be obviously improved and Therapeutic action).

Pharmacological experiment is deployed by classical atherosclerosis animal model, result of study shows that dendrophnol can Significantly reduce total plasma cholesterol and low-density lipoprotein cholesterol level, increasing high density lipoprotein cholesterol levels, significantly Deposition of the cholesterol in vascular wall inner membrance is reduced, suppresses atherosclerosis plaque forming, Aortic Plaque expression of inflammatory cytokines is reduced. The experimental result confirms the dendrophnol preventive and therapeutic action definite to atherosclerosis.The present invention medicine, toxicity compared with Small, security is higher.

Brief description of the drawings

Fig. 1 control groups, model group, dendrophnol group, Atorvastatin group total plasma cholesterol, triglycerides, low-density lipoprotein White cholesterol and HDL-C testing result.⁺⁺P<0.01,⁺⁺⁺P<0.001 is compared with control group；^***P <0.001 Compared with model group.

Fig. 2 control groups, model group, dendrophnol group, Atorvastatin group aortic root atheromatous plaque oil red O stain result 。⁺⁺⁺P<0.001 is compared with control group；^**P<0.01,^***P<0.001 is compared with model group.

Fig. 3 control groups, model group, dendrophnol group, Atorvastatin group sustainer substantially oil red O stain result.⁺⁺⁺P<0.001 Compared with control group；^**P<0.01,^***P<0.001 is compared with model group.

Adhesion molecule1 (ICAM-1), vascular cell adhesion molecule-1 between Fig. 4 model groups and dendrophnol group aorta cells And monocyte chemoattractant protein-1 (MCP-1) mRNA expressions of results (VCAM-1).^*P<0.05,^**P<0.01,^***P<0.001 and mould Type group compares.

Embodiment

According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real Apply the content described by example and be merely to illustrate the present invention, without should be also without limitation on sheet described in detail in claims Invention.

Embodiment 1：Dendrophnol is to Atherosclerosis Model lipid of mice and the influence of Aortic Plaque formation.

1st, experimental animal：ApoE^-/-Mouse, 5-7 weeks, male.

2nd, medicine is prepared：From piece of the dendrophnol content for prevention or the treatment for atherosclerosis of 25mg/ pieces Agent (in addition to dendrophnol, remaining composition is the customary adjuvant for preparing tablet, such as starch, Icing Sugar, lactose, dextrin, maltodextrin and Amylum pregelatinisatum), ultrasonic disperse is in 0.5% sodium carboxymethylcellulose (CMC-Na), and for gastric infusion, dosage is 25mg/kg。

3rd, laboratory apparatus：High speed freezing centrifuge, ABI Step one quantitative real time PCR Instruments, freezing microtome, enzyme mark Instrument, light microscope etc..

Experimental method：

(1) it is grouped and handles：Animal adapted to environment after 1 week, was randomly divided into control group, model group, dendrophnol group and Ah cutting down Statin group, every group 8.Control group gives normal diet, and main component is protein (24.4%), carbohydrate And grease (12.3%) (63.3%).Model group, dendrophnol group, Atorvastatin group give high lipid food, and main component is albumen Matter (23%), carbohydrate (45%), grease (20%) and cholesterol (1.25%).While model is set up, dendrophnol Gavage gives dendrophnol (25mg/kg) and Atorvastatin (10 mg/kg) respectively daily for group and Atorvastatin group.Control group and mould 0.5% isometric sodium carboxymethylcellulose of the daily gavage of type group.Administration time is identical with setting up the disease model time, is 18 Week.

(2) blood sample is collected, for detecting biochemical indicator：T-CHOL, triglycerides, LDL-C And HDL-C.

(3) animal puts to death separation sustainer and heart on ice platform, oil red O stain, the heart after the ethanol of active pulse classic 70% is fixed It is dirty to be embedded through OCT, freezing microtome section (8 μM), oil red O stain, the toning of 60% isopropanol, haematoxylin redyeing, hydrochloride alcohol Differentiation, flowing water is anti-blue.Micro- sem observation sustainer entirety and aortic root atheromatous plaque, using Image J software statistics masters Atherosclerotic plaque area accounts for aortic tunica intima gross area percentage and aortic root atheromatous plaque area.

(4) TRIZOL extraction models group and dendrophnol group sustainer total serum IgE, reverse transcription synthesis cDNA, using real-time are used Quantitative fluorescent PCR (qPCR) detection intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and list Monocyte chemoattractant protein -1 (MCP-1) mRNA expressions.Primer sequence is as follows, ICAM-1 (F：5'- GGAAGGGAGCCAAGTAACTGTGAAG-3', R：5'-GAGCGGCAGAGCAAAAG AAGC-3'), VCAM-1 (F：5'- TGTAGTTCTTTGACAGTCTCCCT-3', R：5'-GTTTGGCTCCA GACATTTACC-3'), MCP-1 (F：5'- GTGCTGACCCCAAGAAGGAATG-3', R：5'-TGAG GTGGTTGTGGAAAAGGTAGTG-3'), GAPDH (F：5'- AAGAAGGTGGTGAAGCAGG-3', R：5'-GAAGGTGGAAGAGTGGGAGT-3').Internal reference used is GAPDH in experiment. The total system of reaction is 20 μ l, including：2 μ l cDNA, 1 μ l primer mixed liquors, 10 μ L iTaq^TM Universal Green Supermix, DEPC water complements to 20 μ l.Response procedures are：Step 1：95℃5min；Step 2：95℃15s； Step 3：60℃1min；Step 2-Step 3：40 circulations.According to 2^-△△CtMethod calculates gene relative expression quantity.

4th, experimental result：

As shown in Figure 1：Compared with control group, model group ApoE^-/-Mice plasma T-CHOL, triglycerides, low-density Lipoprotein cholesterol level is significantly raised, and High-density Lipoprotein-cholesterol is without significant difference；Compared with model group, dendrophnol Significantly reduce ApoE^-/-Mice plasma T-CHOL and low-density lipoprotein cholesterol level, raise ApoE^-/-Mice plasma is highly dense Lipoprotein cholesterol level is spent, triglyceride levels do not make significant difference, illustrate that dendrophnol can significantly improve high fat diet induction Metabolism disorder of blood lipid, and part index number is better than positive control medicine Atorvastatin.

As shown in Figure 2：Compared with control group, model group ApoE^-/-Mouse aorta root atheromatous plaque area significantly increases Plus, illustrate that atherosclerosis mouse model is successfully established.Compared with model group, dendrophnol group ApoE^-/-Mouse aorta root Atheromatous plaque area is significantly reduced, and points out dendrophnol to substantially reduce the aortic root atheromatous plaque shape of high fat diet induction Into.

As shown in Figure 3：Compared with control group, model group ApoE^-/-Mouse aorta atheromatous plaque area is accounted in sustainer Film gross area percentage is significantly raised, and further confirmation atherosclerosis mouse model is successfully established.Compared with model group, stone Dry measure used in former times phenol group ApoE^-/-Mouse aorta atheromatous plaque area accounts for aortic tunica intima gross area percentage and significantly reduced, and points out dendrophnol The aorta atheromatous plaque that high fat diet induction can be substantially reduced is formed.

As shown in Figure 4：Compared with model group, dendrophnol group ApoE^-/-Mouse aorta ICAM-1, VCAM-1 and MCP- 1mRNA expressions are significantly reduced, and are shown that dendrophnol can effectively suppress Aortic Plaque expression of inflammatory cytokines, are mitigated actively Arteries and veins plaque inflammation reacts.

Show that dendrophnol is for atherosclerosis mould by blood parameters, oil red O stain and qPCR testing results Type mouse aorta atherosclerosis plaque forming has obvious inhibitory action, and it is disorderly that its therapeutic action may be by improvement blood lipid metabolism Disorderly with suppression Aortic Plaque inflammatory reaction.

Embodiment 2：

By dendrophnol according to conventional formulation method, water and appropriate solubilizer (polyethylene glycol 400) dissolving, packing are added, is gone out Bacterium, is prepared into atherosclerosis prevention or treatment oral liquid that dendrophnol content is 25mg/mL；

By dendrophnol according to conventional formulation method, soft capsule material selection gelatin and sorbierite, dendrophnol content is prepared into Atherosclerosis prevention or treatment capsule for 25mg/；

By dendrophnol according to conventional formulation method, excipient cyclodextrin is added, is well mixed, granulation, tabletting is prepared into stone Dry measure used in former times phenol content is the atherosclerosis prevention and treatment tablet of 25mg/ pieces.

Embodiment 3：Dendrophnol safety evaluatio.

Acute toxicity test in mice uses single oral fixed dosage method：Select 5,50,500 and 2000mg/kg tetra- Fixed dosage, each 10 Kunming mouses of dosage, 5-7 weeks, male and female half and half.Mouse is administered preceding fasting in 12 hours and can't help water.It is real Mouse is administered in gavage mode in testing, Continuous Observation 2 weeks after administration.As a result find each dosage group of dendrophnol without mouse Death, in addition to mild toxicity reaction occurred in 24 hours in high dose group (2000mg/kg) some animals, remaining each dosage group is small Mouse does not find toxic reaction symptom, illustrates that dendrophnol toxicity is smaller, security is higher.

SEQUENCE LISTING

<110>Hefei Teachers College

<120>Application of the dendrophnol and combinations thereof in prevention and treatment atherosclerosis medicine is prepared

<130> SG20170520

<160> 8

<170> PatentIn version 3.5

<210> 1

<211> 25

<212> DNA

<213> Artificial Sequence

<220>

<223> ICAM-1F

<400> 1

ggaagggagc caagtaactg tgaag 25

<210> 2

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> ICAM-1R

<400> 2

gagcggcaga gcaaaagaag c 21

<210> 3

<211> 23

<212> DNA

<213> Artificial Sequence

<220>

<223> VCAM-1F

<400> 3

tgtagttctt tgacagtctc cct 23

<210> 4

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223> VCAM-1R

<400> 4

gtttggctcc agacatttac c 21

<210> 5

<211> 22

<212> DNA

<213> Artificial Sequence

<220>

<223> MCP-1F

<400> 5

gtgctgaccc caagaaggaa tg 22

<210> 6

<211> 25

<212> DNA

<213> Artificial Sequence

<220>

<223> MCP-1R

<400> 6

tgaggtggtt gtggaaaagg tagtg 25

<210> 7

<211> 19

<212> DNA

<213> Artificial Sequence

<220>

<223> GAPDH-F

<400> 7

aagaaggtgg tgaagcagg 19

<210> 8

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223> GAPDH-R

<400> 8

gaaggtggaa gagtgggagt 20

Claims

1. application of the dendrophnol and combinations thereof in prevention and treatment atherosclerosis medicine is prepared.

2. application of the dendrophnol and combinations thereof in prevention of arterial anti-atherosclerotic agent is prepared.

3. application of the dendrophnol and combinations thereof in treatment atherosclerosis medicine is prepared.

4. according to any described application of claims 1 to 3, it is characterised in that dendrophnol is equipped with to the auxiliary material system of corresponding formulation Into oral liquid, injection, tablet, pill, dispersant, capsule, soft capsule, dripping pill, granule, solution, suspending agent, emulsion or Microcapsules.