CN103665180B - A kind of glycan of morinda root oligosacchride 6 prepare resist myocardial ischemia and reperfusion injury medicine in application - Google Patents
A kind of glycan of morinda root oligosacchride 6 prepare resist myocardial ischemia and reperfusion injury medicine in application Download PDFInfo
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Abstract
The invention discloses the application of Radix Morindae Officinalis extract and the glycan of morinda root oligosacchride 6 in preparation resists myocardial ischemia and/or Reperfusion injury injures promotion Therapeutic Angiogenesis medicine.Described myocardial ischemia and reperfusion injury is myocardium when hemoperfusion stops or be bad, that is, undergoes ischemic, anoxia-induced apoptosis, and re-establish the damage again brought after circulation, recovery blood supply to body.There is protective effect to rat in vivo Model of Myocardial Ischemia-Reperfusion Injury and the anoxia reoxygenation injury of neonatal cardiac myocytes model of ex-vivo purified culture; the generation that myocardial infarction Mian Ji ﹑ mitigate Reperfu- sion phase arrhythmia cordis can be reduced significantly; effectively the form of protection cardiac muscle cell, mitigates damage of the hypoxia-reoxygenation to cardiac muscle cell.Described promotion Therapeutic Angiogenesis is that the therapeutic action for including some drugses by the stimulation of certain methods increases functional coronary arterial tree or collateral, reaches recovery ischemic myocardium blood supply, improves the purpose of cardiac function.
Description
Technical field
The present invention relates to the medicinal application field of plant extracts, and in particular to morinda root oligosacchride resists myocardial ischemia in preparation
And the application in reperfusion injury medicine.
Background technology
Coronary atherosclerotic heart disease (coronary heart disease, CAD), abbreviation coronary heart disease, also referred to as
It is the common angiocardiopathy of the mankind for ischemic heart disease, is topmost adult's lethal angiocardiopathy.Coronary heart disease is led
The acute myocardial infarction of cause is to cause the main cause that cardiovascular disease incidence rate is high, the death rate is high at present.Incidence of coronary heart disease is held
It is continuous to increase, and rejuvenation trend is obvious.Coronary heart disease is clinically presented with serious myocardial ischemia, seriously threatens the mankind
Health.The death rate and long-term prognosis of patients with coronary heart disease are related to the left ventricular function that myocardial damage area is determined, and myocardium
Damaged area is determined by the degree and time of myocardial ischemia.Cardiac muscle blocks more than 30min in blood supply, will cause each
The change of organelle ultra microstructure and dysfunction, cause the irreversible damage of cardiac muscle cell even dead.Myocardial ischemia is damaged
Wound is aggravated with the extension of Ischemia Time.
Thrombolysis, coronary artery artery bypass and calcification score etc. are the effective ways for treating acute myocardial infarction AMI, these methods
Obstruction Coronary recanalization, but the timely Reperfu- sion of myocardial ischemia can be made, although partial injury but the not yet downright bad heart can be saved
Myocyte, it is also possible to aggravate the damage of ischemic myocardium, shows as the necrosis quickening of cardiac muscle cell, cellular swelling, muscle fibers contract
The formation of band, myocardium internal haemorrhage and without perfusion phenomenon, it is also possible to produce serious reperfusion arrhythmia.This coronary heart disease hair
Occur in treating myocardial ischemia-reperfusion injury (myocardial ischemical reperfusion injury,
MIRI the recovery of survival rate and Long-term Cardiac Function instant after coronary artery revascularization) has been had a strong impact on.
Morinda officinalis (Radix morinda officinalis, RMO), is madder wort (Rubiaceae), is China
One of four famous great Nan medicines, are also the rare medicinal herbs of foreign exchange earning.Morinda officinalis has a tonifying kidney and strengthening yang, strengthening the bones and muscles, wind-damp dispelling it
Effect, toxicological experiment proves that it is a kind of safe drugs.There is document report, the content of 5~7 years raw Morinda officinalis total reducing sugars can exceed
50%, it is the important component of Morinda officinalis chemical composition.Activity research shows, Morinda officinalis carbohydrate content have immunological regulation,
Antitumor, antidepression, resisting stress, anti-inflammatory, anti-oxidant, the anti-ageing effect of waiting for a long time.
The content of the invention
The present invention is particularly to Radix Morindae Officinalis extract is extracted from madder wort Morinda officinalis (Morinda officinalis
How the bioactivity of the morinda root oligosacchride (Morindae officinalis oligosaccharides, MOO) of dry root)
Studied.Applicant is found surprisingly that, Radix Morindae Officinalis extract resist myocardial ischemia and/or reperfusion injury in have it is obvious
Activity.Its active component is followed the trail of find its Ischemic myocardium and reperfusion injury, promote angiogenesis effect it is effective into
It is divided into the mixture of inulin type oligosaccharide, 4,5,6 glycan are to promoting anoxia/reoxygenation wherein in morinda root oligosacchride monomer
Human umbilical vein endothelial cells (HUVEC) propagation and tubule formation effect afterwards is most obvious, and mutually collaboration is acted between three.
The glycan molecule formula of morinda root oligosacchride 4：C24H42O21, molecular weight：666.58, CAS accession number：13133-07-8, chemistry
Title：Nystose Nystose, structural formula I；Abridge Hex4.
The glycan molecule formula of morinda root oligosacchride 5：C30H52O26, molecular weight：828.72, CAS accession number：59432-60-9, chemistry
Title：GF4 1F-fructofuranosylnystose, formula II；Abridge Hex5.
The glycan molecule formula of morinda root oligosacchride 6：C36H62O31, molecular weight：990.86, chemical name：Six sugar Fru β (2- of sugarcane fruit
1)-[Fru β (2-1)] 4- α (2-1) Glc, formula II I, abridge Hex6.
Our research finds that the glycan of Morinda officinalis 4,5,6 can be reduced significantly myocardial infarction area first, mitigates anoxic and answers
Damage of the oxygen to cardiac muscle cell, reduces the form of the generation of Reperfu- sion phase arrhythmia cordis, effectively protection cardiac muscle cell, promotes treatment
Property angiogenesis, its therapeutic effect be better than Diaoxinxue Kang and HMP.Above-mentioned myocardial ischemia and reperfusion injury are the hearts
Flesh undergo ischemic, anoxia-induced apoptosis, and re-establish circulation, recover after blood supply to body when hemoperfusion stops or be bad
The damage again brought.
Based on above-mentioned discovery, exist it is an object of the invention to provide the Radix Morindae Officinalis extract particularly glycan of Morinda officinalis 4,5,6
Prepare the new application in terms of New Effective Drugs Against Myocardial Ischemia-Reperfusion Injury.Described promotion Therapeutic Angiogenesis is by some sides
The therapeutic action that the stimulation of method includes some drugses increases functional coronary arterial tree or collateral, reaches recovery ischemic myocardium
Blood supply, improves the purpose of cardiac function.
Resisted myocardial ischemia the invention provides Radix Morindae Officinalis extract in preparation, reperfusion injury or promote Therapeutic vascular new
Application in crude drug thing.
Further, the Radix Morindae Officinalis extract is extracted from the dry root of madder wort Morinda officinalis.
Further, resisted myocardial ischemia the invention provides morinda root oligosacchride in preparation, reperfusion injury or promote treatment
Application in property angiogenesis medicament.Preferably, the morinda root oligosacchride is included in structural formula I, formula II and formula II I
The one or more of shown compound.
Further, the invention provides a kind of pharmaceutical composition, it is characterised in that bar is included in described pharmaceutical composition
Halberd day oligosaccharide compositions.Preferably, the morinda root oligosacchride includes change shown in said structure Formulas I, formula II and formula II I
The one or more of compound.Pharmaceutical composition, its formulation can be tablet, pill or capsule, injection etc..
Further, the invention provides a kind of preparation method of the glycan of morinda root oligosacchride 4,5,6, comprise the following steps：
1), Radix Morindae Officinalis extract is dissolved in the water, is then freeze-dried；
2) pyridine, is added in freeze-dried thing, and in 0 DEG C of following stirring, side adds acetic anhydride；
3), stir after 24h, be concentrated under reduced pressure under chamber state；
4), residual fraction adds CHCl3, cleaned with saturation soda water, saturated aqueous common salt；
5) after, being dried with magnesium sulfate, then removal CHCl is depressurized3；
6), purified respectively with compound method for refining, expansion solvent is toluene：Ethyl acetate=2:1→1:1→1:2→1:3
→1:5, respectively obtain the glycan monomer of acetylation morinda root oligosacchride 4,5,6；
7), magnetic resonance detection determines the resulting glycan monomer structure of acetylation morinda root oligosacchride 4,5,6 respectively；
8), the glycan monomer of acetylation morinda root oligosacchride 4,5,6 is deacetylated
The glycan monomer of acetylation morinda root oligosacchride 4,5,6 is dissolved in MeOH, CH is added3ONa, is stirred at room temperature
After 2.5h, add Dowex 50W x 2 [H+] and neutralized, after resin filter, decompression removes solvent, by residual fraction
It is dissolved into H2In O, by 10ml Bio-Gel P-2 (polyacrylamide gel) carry out desalination, it is freeze-dried after, obtain white
Solid；
9), white solid is miscellaneous through Matrix-assisted laser desorption ionization (MALDI-TOF-MS) technology for detection
Matter content and the molecular weight for determining the glycan monomer of morinda root oligosacchride 4,5,6.
Wherein, the myocardial ischemia-reperfusion injury, be cardiac muscle hemoperfusion stop or it is bad when, that is, undergo ischemic,
After anoxic, circulation is re-established, the damage that blood supply is brought after recovering to body.
The glycan of Morinda officinalis 4,5,6 of the present invention is extracted from madder wort Morinda officinalis (Morinda officinalis
How dry root).
Medicine of the present invention refers to, using the glycan of Morinda officinalis 4,5,6 as main active, be made with pharmaceutical acceptable carrier
Medically acceptable preparation, such as injection, tablet, pill or capsule.
Medicine of the present invention, its dose is in terms of the glycan of Morinda officinalis 4,5,6：0.5g/ times or 1.5g/ days.
The glycan of Morinda officinalis 4,5,6 of the present invention is available directly by being commercially available, and can also be obtained by plant extract.
The present invention passes through：1. isolated cells experimental system：Neonatal rat myocyte anoxia/reoxygenation, Apoptosis, people
Breed after huve cell (HUVEC) anoxia/reoxygenation, migration and tubule are formed；2. Integral animal experiment system：Nothing
Invasive, invasive myocardial ischemia/Infarction Model；Research of the glycan of Morinda officinalis 4,5,6 to cardiovascular protective effect is carried out.
The beneficial effects of the present invention are：
1st, the invention provides the morinda root oligosacchride new application based on 4,5,6 glycan, it has very well to cardiovascular system
Protective effect.
On the basis of a large amount of pertinent literatures both at home and abroad are consulted, the work of cardiovascular system is protected to the glycan of Morinda officinalis 4,5,6
With being studied, as a result confirm：1) to rat in vivo Model of Myocardial Ischemia-Reperfusion Injury and the suckling mouse of ex-vivo purified culture
Myocytes Anoxia reoxygenation model has protective effect, can be reduced significantly myocardial infarction Mian Ji ﹑ and mitigate the Reperfu- sion phase rhythm of the heart
The form of not normal generation, effectively protection cardiac muscle cell, mitigates damage of the hypoxia-reoxygenation to cardiac muscle cell；2) the rat heart can be improved
Myocyte's antioxidase (SOD, LDH, GSH-Px) level, lipid peroxidation when mitigating rat myocardial cell ischemical reperfusion injury
Degree of injury, reduces lipid peroxidation product MDA generation；3) iNOS expressions when can improve rat myocardial ischemia and reperfusion
And NO yield, improve cardiac muscle cell Ca2+- ATPase and Na+-K+- ATPase activity；4) dependency basis of regulating cell apoptosis is passed through
Because of expression, it can significantly mitigate the apoptosis of cardiac muscle cell；5) by reduce during cardiomyocyte injury induced by hypoxia/reoxygenation ET-1 contents and
The secretion of TNF-α, IL-1 β, IL-6, significantly improves inflammatory reaction degree during Myocytes Anoxia reoxygenation；6) it can improve
The haemodynamics of blood stasis rat, suppresses platelet aggregation；7) Morinda officinalis sugar chain can remarkably promote myoblast proliferation and centripetal
Myoid cells breaks up, and strengthens the PCNA positive reactions of sarcoblast karyon, improves TGF-β 1, TGF-β 2, GATA- in sarcoblast
The expression of 4 genes and albumen, up-regulation information conduct factors Smad4 expression, according to dosage bidirectional modulation Idiotype signal transduction because
Sub- p-Smad2/3 and Smad7 expression.8) angiogenesis can be effectively facilitated, significantly mitigates damages of the H/R to vascular endothelial cell
Wound, suppresses Apoptosis, effectively facilitates endothelial cell migration and the vascularization of H/R damages, improves ischemic myocardium Microvascular density
Degree, vegf protein in notable induction of vascular Newborn Process, bFGF albumen, Flt-1 albumen, TRPC6 albumen, ERK1/2 albumen and
The expression of PDGF-B albumen.Regulate and control the major signaling molecule protein expression such as Notch1/Jagged1, thus reduce Notch signals and
The expression of pten protein.9) big conductance calcium-activated potassium (BKCa) passage on endothelial cellular membrane can be activated.10) Morinda officinalis sugar chain
There is doses correlation to the protective effect of cardiovascular system.
2nd, the glycan of Morinda officinalis 4,5,6 that uses of the present invention is extracted from pure natural raw material, and extracting method is simple, chemical constitution and
Clearly, property is stable for molecular weight, and recovery rate is high, quality controllable.
Brief description of the drawings
Fig. 1 is cardiac muscle cell's morphological observation (× 40), and A is normal myocardial cells, and B is H/R groups, and C is the H/R+ ground heart difficult to understand
Blood health group, D is H/R+4 glycan groups, and E is H/R+5 glycan groups, and F is H/R+6 glycan groups；
Fig. 2 is Myocardial ultrastructure observation (× 5000), and A is normal myocardial cells, B is H/R groups, and C is H/R+
Sodium Ferulate group difficult to understand, D is H/R+4 glycan groups, and E is H/R+5 glycan groups, and F is H/R+6 glycan groups；
Fig. 3 is myocardial infarction substantially tissue specimen, and A is normal myocardium, and B is MI groups, and C is MI+ Diaoxinxue Kang groups, and D is
MI+4 glycan groups, E is MI+5 glycan groups, and F is MI+6 glycan groups；
Fig. 4 is ischemic myocardium microvessel density (MVD) detection, and A is normal myocardium, and B is MI groups, and C is that MI+ Moschus protects the heart
Ball group, D is MI+4 glycan groups, and E is MI+5 glycan groups, and F is MI+6 glycan groups；
Fig. 5 is that HUVEC anoxia/reoxygenations (H/R) breed (× 10) afterwards, and A is normal group, and B is H/R groups, and C is H/R+ musk deers
Fragrant heart pill group, D is H/R+4 glycan groups, and E is H/R+5 glycan groups, and F is H/R+6 glycan groups；
Fig. 6 HUVEC migrate (× 10), and A is normal group, and B is H/R groups, and C is H/R+ HMP groups, and D is poly- for H/R+4
Sugared group, E is H/R+5 glycan groups, and F is H/R+6 glycan groups；
Fig. 7 HUVEC tubules formation (× 10), A is normal group, and B is H/R groups, and C is H/R+ HMP groups, and D is H/R+
4 glycan groups, E is H/R+5 glycan groups, and F is H/R+6 glycan groups；
Fig. 8 HUVEC tubules formation (× 10), A is normal group, and B is H/R groups, and C is H/R+ HMP groups, and D is H/R+
MOO groups, E is H/R+Hex4, and F is H/R+Hex5, and G is H/R+Hex6；
Embodiment
The experiment material such as biochemical reagents, experimental animal and cell used can be obtained by commercially available channel in the present invention.
Morinda officinalis medicinal material (Radix morinda officinalis) is purchased from Guangdong Province Deqing County medicinal material company, through Henan College Of Traditional Chinese Medicine
Pharmaceutical college is accredited as Grade A；Morinda root oligosacchride mixture (Morinda officinalis oligosaccharides) is used
This area conventional method is prepared and determined；SD/Wistar rats are purchased from Zhengzhou University's Experimental Animal Center；Human umblilical vein endothelial is thin
Born of the same parents (HUVEC) are purchased from Wuhan University's cyropreservation center, and above-mentioned material can also be bought by other commercial sources and be obtained.
Embodiment 1
The preparation of the glycan monomer of morinda root oligosacchride 4,5,6：
(1) 10g Radix Morindae Officinalis extracts are dissolved in 80ml H2In O, then make its freeze-dried.
(2) 100ml pyridine is added in 8.3g freeze-dried thing, and side is stirred at a temperature of 0 DEG C, side is added
100ml acetic anhydride.
(3) stirred under chamber state after 24h, unnecessary medium is removed with decompression method.
(4) residual fraction adds CHCl3, cleaned with saturation soda water, saturated aqueous common salt.
(5) after being dried with magnesium sulfate, then removal CHCl is depressurized3。
(6) (expansion solvent is purified respectively with compound method for refining：Toluene：Ethyl acetate (Toluene:Ethyl
Acetate)=2:1→1:1→1:2→1:3→1:5).Extract 623mg acetylation Hex4 monomers (Hex4perAc).Carry
Take out 623mg acetylation Hex5 monomers (Hex5perAc).Extract 74mg acetylation Hex6 monomers (Hex6perAc).
(7) Hex4perAc, Hex5perAc, Hex6perAc structure are determined with magnetic nuclear resonance method.
(8) Hex4perAc, Hex5perAc, Hex6perAc are deacetylated：
100mg Hex4perAc is dissolved in 5mL MeOH, 18mg 0.3eq CH is added3Ona.Under greenhouse
Stir after 2.5h, add Dowex 50W x 2 [H+] and neutralized.After resin filter, decompression removes solvent.Will residual
It is partly dissolved H2In O, pass through (Bio-Gel P-2:10ml) carry out desalination.After freeze-dried, white solid can be obtained
(57mg)。
100mg Hex5perAc is dissolved in 5mL MeOH, 18mg0.3eq CH is added3Ona.Under greenhouse
Stir after 2.5h, add Dowex 50W x 2 [H+] and neutralized.After resin filter, decompression removes solvent.Will residual
It is partly dissolved H2In O, pass through (Bio-Gel P-2:10ml) carry out desalination.After freeze-dried, white solid can be obtained
(57mg)。
74mg Hex6perAc is dissolved in 5mL MeOH, 12mg0.3eq CH is added3Ona.Stirred under greenhouse
Mix after 2.5h, add Dowex 50W x 2 [H+] and neutralized.After resin filter, decompression removes solvent.By residual part
Divide and be dissolved into H2In O, pass through (Bio-Gel P-2:10ml) carry out desalination.After freeze-dried, white solid (39mg) can be obtained.
(9) white solid is miscellaneous through Matrix-assisted laser desorption ionization (MALDI-TOF-MS) technology for detection
Matter content and the molecular weight for determining the glycan monomer of morinda root oligosacchride 4,5,6.
Hex4：Purity more than 95% (NMR)；Preserve 4 DEG C；Molecular weight 666.578；
Hex5：Purity more than 95% (NMR)；Preserve 4 DEG C；Molecular weight 828.7183；
Hex6：Purity more than 95% (NMR)；Preserve 4 DEG C；Molecular weight 990.859.
Embodiment 2
(1) in vitro endothelial cell and Integral animal experiment
<1>In the experimental system of Neonatal Mouse cardiac muscle cell, using newborn (1-3d) SD/Wistar rats, core dirty
Do primary myocardial cell culture, be divided into Normal group, Hypoxia/Reoxygenation Injury (H/R) model group, H/R+ positive drugs (it is difficult to understand
Sodium Ferulate, 0.7gL-1) control group, H/R+Hex4 (42nmolml-1)、Hex5(30nmol·ml-1)、Hex6(22nmol·
ml-1) each group.Using cardiac muscle cell's form and Myocardial ultrastructure as main evaluation index, the glycan pair of Morinda officinalis 4,5,6 is observed
The protective effect of cardiac muscle cell.
<2>In the experimental system of Human umbilical vein endothelial cells (HUVEC) model of angiogenesis, be divided into Normal group,
H/R model groups, H/R+ positive drugs (HMP, 2gL-1) control group, H/R+Hex4 (42nmolml-1)、Hex5
(30nmol·ml-1)、Hex6(22nmol·ml-1) each group.Tube chamber knot is arranged in vascular endothelial cell proliferation and endothelial cell
Structure is main evaluation index, equal higher than Normal group into luminal structure number with cell arrangement with anoxia model group cell propagation
Two standard deviations of value are used as the successful evaluation criterion of model.Different medicines are given respectively in 12h after model success.
1. 6h, 12h, 24h, 48h, 72h determine the increasing of each group vascular endothelial cell with fluidic cell method respectively after administration
Grow；2. each group cell generation cycle is analyzed with flow cytometric methods.3. tested and small tubular using the migration of transwell cells
The effect for promoting the propagation of endothelial cell into the glycan of Germicidal efficacy Morinda officinalis 4,5,6 and inducing it to be differentiated to form tube chamber.
(2) Integral animal experiment
<1>Non-invasive myocardial infarction and ischemia model：From SD/Wistar rats, animal is randomly divided into：Blank (physiological saline)
Control group, isoprel (Isoproterenol) Chronic myocardium ischemia damage model group, model+positive drug (the heart difficult to understand
Blood health, 28mgkg-1·d-1) control group, model+Hex4 (24mgkg-1·d-1)、Hex5(12mg·kg-1·d-1)、Hex6
(2.8mg·kg-1·d-1) each group.In addition to blank control group intraperitoneal injection of saline, remaining each group uses multiple isopropyl kidney
Upper parathyrine be injected intraperitoneally Methods of Preparing Myocardial chronic ischemia damage model, by decision model of the electrocardiogram ST-T changes successfully according to
According to.In starting in 24h to give pharmaceutical intervention after model success, (blank group, model group gavage give the physiology of isometric(al) to each group
Salt solution, each administration group gavage gives the medicine of corresponding isometric(al) various dose).
<2>Invasive myocardial infarction model：From SD/Wistar rats, be randomly divided into false ligation group, Infarction Model group,
Model+positive drug (HMP group 30mgkgd) control group, model+Hex4 (24mgkg-1·d-1)、Hex5
(12mg·kg-1·d-1)、Hex6(2.8mg·kg-1·d-1) each group.Except false ligation group opens a threading after chest separation coronary artery
Outer, remaining each group following coronary artery occlusion left anterior descending branch at pulmonary conus and the slightly lower 1~2mm in left auricle of heart boundary is not ligatured,
Myocardial infarction model is prepared, is bleached with ligaturing cardiac muscle under position, decrease of beating and electrocardiogram the ST sections of back of a bow occur and substantially lifted upwards
A height of modeling Success Flag.In starting pharmaceutical intervention after modeling in 24h, (false ligation group, Infarction Model group are injected intraperitoneally each group
The physiological saline of isometric(al) is given, each administration group intraperitoneal injection gives corresponding isometric(al) various dose medicine).
Animal, cardiac muscle materials are put to death after pharmaceutical intervention 6w：1. specimens paraffin embedding slices, the general Histomorphological of row；②
Microvessel density (MVD) in HE dyeing detection each group ischemic myocardium in rat, the glycan of Morinda officinalis 4,5,6 promotes from integral level
The feature of angiogenesis.
Experimental result is as shown in the figure：
Fig. 1 is Morinda officinalis 4,5,6 influence pair neonatal rat myocardial cell form：It can be seen that the normal heart under inverted microscope
After myocyte cultivates 3 days, pseudopodium is stretched out in the growth of cell cluster, and refractivity is strong, and beating is obvious.Variation is presented in cellular morphology,
Such as circular, fusiformis and vertebra shape (figure A).Cardiac muscle cell's pseudopodium shortens or disappeared after anoxia/reoxygenation, and refractivity declines, and beating subtracts
Weak or disappearance (figure B).Diaoxinxue Kang group cardiac muscle cell form has some improvement (figure C) compared with anoxia/reoxygenation group.Hex4、
Each administration group cardiac muscle cell form of Hex5, Hex6 be improved significantly (figure D, E, F).
Fig. 2 is Morinda officinalis 4,5,6 influence pair neonatal rat myocardial cell ultra microstructure：Cardiac muscle cell's ultra micro under transmission electron microscope
Structure shows that normal cell culture group ultra microstructure is complete, clear, and organelle is more, and nuclear membrane surface is smooth, chromatin in core
Loose, density is homogeneous (figure A).The visible cell space of Hypoxia-reoxygenation model group diminishes, nuclear membrane shrinkage, and chromatic agglutination has uneven side
Collect phenomenon, part chromatin gathers under complete nuclear membrane to be changed in demilune sample, and nuclear hyperchromatism, pyknosis, mitochondria has vacuole change
Property etc. apoptosis representative configuration feature (figure B).Diaoxinxue Kang group cell ultrastructure obtains different journeys compared with anoxia/reoxygenation group
The improvement of degree, but still have the slight side collection of chromatin, the morphological change such as nuclear hyperchromatism (figure C).Each administration group of Hex4, Hex5, Hex6
Cell ultrastructure is close normal (figure D, E, F).
Fig. 3 is Morinda officinalis 4,5,6 influence pair rat myocardial infarction model：It is visible in myocardial infarction substantially tissue specimen, and just
Normal group (figure A) is compared, and myocardial infarction model group cardiac muscle is in large area transmural ischemic necrosis (figure B).Diaoxinxue Kang group cardiac muscle
Ischemic necrosis area has mitigated (figure C) compared with model group.Each administration group myocardial ischemia and necrosis area of Hex4, Hex5, Hex6 and journey
Degree substantially mitigates (figure D, E, F) compared with model group.
Fig. 4 is the influence of Morinda officinalis 4,5,6 pairs of rat heart muscle blood vessel hyperplasias and microvessel density：Visible cardiac muscle is cut under light microscopic
The blood capillary of new life can be seen in false ligation group, model group, HMP group, each administration group of Hex4, Hex5, Hex6 in piece
Pipe；But the visible blood capillary proliferation of false ligation group substantially (does not scheme A).The visible a small amount of blood capillary proliferation (figure B) of model group.Moschus
The capillary (figure C, D, E, F) of heart pill group and the visible most hyperplasia of Hex4, Hex5, Hex6 each group.Compared with false ligation group,
Model group rats MVD (microvessel density) significantly increases (P ＜ 0.05).Compared with model group, HMP group and Hex4,
Hex5, Hex6 each group can dramatically increase MVD (P ＜ 0.05).
Fig. 5 is Morinda officinalis 4,5,6 influence pair cell proliferation of human umbilical vein：Compared with normal group (figure A), anoxic is multiple
Oxygen injury group HUVEC cell proliferating number amounts are less (figure B).Compared with model group, HMP group cell proliferating number amount increases,
Statistics has significant difference (P ＜ 0.05 scheme C).Compared with HMP group, each medication group of Hex4, Hex5, Hex6
Remarkably promote the propagation after cellular damage (P ＜ 0.05 scheme D, E, F).
Fig. 6 is Morinda officinalis 4, the influence 1 (cell method) of 5,6 pairs of Human umbilical vein endothelial cells migration：With normal group (figure A) phase
Than anoxia/reoxygenation (H/R) model group cell migration number significantly reduces (P<0.05, scheme B).Positive control drug HMP
Group does not show to promote the effect (figure C) of anoxia/reoxygenation endothelial cell migration.Hex4, Hex5, Hex6 each group can substantially increase
Plus after anoxia/reoxygenation cell migration number (P<0.05, figure, D, E, F).
Fig. 7 is Morinda officinalis 4, the influence 2 (scarification) of 5,6 pairs of Human umbilical vein endothelial cells migration：With normal group (figure A) phase
Than anoxia/reoxygenation (H/R) model group cell migration number significantly reduces (P<0.05, scheme B).Positive control drug HMP
Group has the trend (figure C) for promoting anoxia/reoxygenation endothelial cell migration.Hex4, Hex5, Hex6 each group can substantially increase anoxic
Migration number (the P of cell after reoxygenation<0.05, scheme D, E, F).
Fig. 8 is the influence that Morinda officinalis 4,5,6 promotees the formation of Human umbilical vein endothelial cells tubule：Compared with normal group (figure A),
MOO, Hex4, Hex5, Hex6 each group cell culture 6h be it is visible have intercellular interconnection form, 18h is in martrigel tables
Face forms complete tube-like structures, and with the extension of incubation time, luminal structure increases (figure D, E, F, G).Hypoxia-reoxygenation
Damage model group cell just starts to be connected with each other after 18h, and 24h forms complete tube-like structures on matrigel surface and is less than
MOO, Hex4, Hex5, Hex6 each group (figure B).Positive control drug HMP group tubule formation situation be worse than MOO, Hex4,
Hex5, Hex6 each group (figure C).Through computer image analysis software handle obtain tubule formation area show, model group with normally
Group compares significant difference (P<0.05), MOO, Hex4, Hex5, Hex6 each group significant difference (P compared with model group<0.05), mould
Type group indifference compared with HMP group is notable (P ＞ 0.05).
Preferred embodiment of the invention described in detail above.It should be appreciated that the ordinary skill of this area is without wound
The property made work just can make many modifications and variations according to the design of the present invention.Therefore, all technical staff in the art
Pass through the available technology of logical analysis, reasoning, or a limited experiment on the basis of existing technology under this invention's idea
Scheme, all should be in the protection domain being defined in the patent claims.
Claims (1)
1. the glycan of morinda root oligosacchride 6 prepare resist myocardial ischemia, reperfusion injury or promote Therapeutic Angiogenesis medicine in
Using, wherein, the glycan of morinda root oligosacchride 6 is compound shown in formula II I
。
Priority Applications (1)
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1289592A (en) * | 1999-09-24 | 2001-04-04 | 中国人民解放军军事医学科学院毒物药物研究所 | Application of oligose treating stress brain injury |
| CN1587271A (en) * | 2004-07-30 | 2005-03-02 | 广州中医药大学 | Process for preparing morinda officinalis total oligosaccharide |
| CN101987198A (en) * | 2009-08-06 | 2011-03-23 | 北京美倍他药物研究有限公司 | Medicinal composition |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1289592A (en) * | 1999-09-24 | 2001-04-04 | 中国人民解放军军事医学科学院毒物药物研究所 | Application of oligose treating stress brain injury |
| CN1587271A (en) * | 2004-07-30 | 2005-03-02 | 广州中医药大学 | Process for preparing morinda officinalis total oligosaccharide |
| CN101987198A (en) * | 2009-08-06 | 2011-03-23 | 北京美倍他药物研究有限公司 | Medicinal composition |
Non-Patent Citations (1)
| Title |
|---|
| 巴戟天醇提取物促大鼠缺血心肌治疗性血管生成的实验研究;杨景柯等;《中国药理学通报》;20100331;第26卷(第3期);第367-371页 * |
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