CN105902583A - Chestnut shell extract and preparation method and application thereof - Google Patents
Chestnut shell extract and preparation method and application thereof Download PDFInfo
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- CN105902583A CN105902583A CN201610338795.6A CN201610338795A CN105902583A CN 105902583 A CN105902583 A CN 105902583A CN 201610338795 A CN201610338795 A CN 201610338795A CN 105902583 A CN105902583 A CN 105902583A
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- epicarpium castaneae
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- 238000000605 extraction Methods 0.000 title claims abstract description 20
- 241001070941 Castanea Species 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 235000014036 Castanea Nutrition 0.000 title claims abstract description 18
- 102000002072 Non-Receptor Type 1 Protein Tyrosine Phosphatase Human genes 0.000 claims abstract description 30
- 108010015847 Non-Receptor Type 1 Protein Tyrosine Phosphatase Proteins 0.000 claims abstract description 30
- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 20
- 208000008589 Obesity Diseases 0.000 claims abstract description 10
- 235000020824 obesity Nutrition 0.000 claims abstract description 10
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract description 8
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 238000000638 solvent extraction Methods 0.000 claims abstract description 3
- 239000003480 eluent Substances 0.000 claims description 18
- 238000000746 purification Methods 0.000 claims description 17
- 230000001629 suppression Effects 0.000 claims description 15
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a chestnut shell extract and a preparation method and application thereof, belonging to the technical field of natural product extraction and application. The preparation method comprises the following steps: extraction: the chestnut shell extract is prepared by using chestnut shells as raw materials and ethanol water solution with the volume percentage concentration of 40-100% as a solvent, performing solvent extraction, and recovering the solvent. The chestnut shell extract has the function of inhibiting protein tyrosine phosphatase 1B (PTP1B), can be used as PTP1B inhibitor, and can be used for treating diabetes, obesity and other complications caused by diabetes, obesity and other complications.
Description
Technical field
The present invention relates to natural product extraction applied technical field, particularly relate to a kind of epicarpium Castaneae extract and
Preparation method and application.
Background technology
At present, often diabetes are divided into insulin-dependent (IDDM, type Ⅰ diabetes mellitus) and non-pancreas clinically
Island element dependent form (NIDDM, type Ⅱdiabetes mellitus) two classes, wherein type Ⅱdiabetes mellitus accounts for 90% in diabetes.
WHO it is expected that due to aged tendency of population, obesity, unsound diet and lack motion life style,
By 2025, the number of diabetics will be risen to 300,000,000 by the 1.35 hundred million of nineteen ninety-five.
The feature of type Ⅱdiabetes mellitus is insulin sensitive tissues, if skeletal muscle, liver, fatty tissue are to insulin
The opposing of effect.Although its concrete mechanism is unclear, but insulin signaling weakening in its pathway
Even block and must have direct relation.
PTPases (protein-tyrosine-phosphatase) multiple links in insulin signaling pathway work, as
By the IR dephosphorylation of self phosphoric acid activation, thus reduce kinase activation；Or by IRS-1, IRS-2,
Protein-tyrosine residue dephosphorylation in the substrate of the Insulin receptor INSRs such as Shc, thus negative regulation insulin action is subject to
Path after body.The enzymatic activity imbalance between tyrosine kinase in specific PTPases and insulin path may
It it is the reason causing type Ⅱdiabetes mellitus insulin resistant.
Therefore, by finding selectively acting inhibitor of PTPases in this path, suppress its activity, add
By force with prolongation insulin signaling, become the new way of more and more valued treatment type Ⅱdiabetes mellitus.
At present, the research of PTP1B (protein-tyrosine-phosphatase 1B) selective depressant achieves necessarily
Progress, but mostly study limitation is in peptides and class peptide compounds, such as: dephosphorylized according to PTP1B
Inhibitor EEDE (F2PMP) M, Glu-F2PMP-F2PMP of substrate sequence design, although these peptides
Compounds has higher selectivity and a stronger inhibitory activity, but they be difficult to permeates cell membranes, with
And the structure of peptides phosphoric acid makes it difficult to become drug candidate compound.
Recently, in the report of non-peptides PTP1B inhibitor, 2-carboxymethoxyl benzoic acid derivative pair
PTP1B has the strongest inhibitory action, it is often more important that, wherein (2S)-2-[4 '-(2-Benzyl-benzofuran)-3-connection
Benzene-4-oxygen]-3-phenyl-propionic compound not only has a strongest selection inhibition, also to reducing ob/ob to PTP1B
Glucose and insulin level in mice plasma have remarkable effect.This is that first case is from the most directly demonstrate,proving
Bright PTP1B inhibitor has evidence (Malamas, the M.S.et al.J.Med.Chem. of anti-diabetic activity
2000,43,1293-1310).This finds efficiently for us from natural resources, high selective little molecule is non-peptide
Class PTP1B inhibitor provides opportunity.
Summary of the invention
Based on this, the present invention finds from natural resources, it is provided that a kind of have the natural of PTP1B inhibitory activity
Composition.
The preparation method of a kind of epicarpium Castaneae extract, comprises the following steps:
Extract: with chestnut shell as raw material, the ethanol water with concentration expressed in percentage by volume as 40%-100% as solvent,
Carry out solvent extraction, recycling design, obtain epicarpium Castaneae extract.
The Semen Castaneae (Castanea mollissima Blume) that the present invention relates to, belongs to Fagaceae Fagaceae Castanea
Nuts plant, has another name called chestnut, Semen Castaneae, great Li etc., is the special product plant of China, main product in Guangdong, Guangxi,
The ground such as Yunnan, Jiangxi.The medicinal part of Semen Castaneae includes root, skin, leaf, flower, outer peel (epicarpium Castaneae), interior fruit
Skin (LIPI), involucre (Involucrum Castaneae or acorn-cup) etc..
Above-mentioned chestnut shell is the outer peel of Semen Castaneae, it is generally recognized that its property of medicine is sweet, puckery, flat, has sending down the abnormal ascending QI, stops
Effect of blood.
The present inventor finds after long-term experimentation, the ethanol extraction of Semen Castaneae outer peel (i.e. epicarpium Castaneae)
Thing have suppression protein-tyrosine-phosphatase 1B (PTP1B) effect, it is possible to using this extract as
PTP1B inhibitor uses, for treating diabetes, obesity and other complication thus caused.
Wherein in an embodiment, in described extraction step, described solvent is that concentration expressed in percentage by volume is
The ethanol water of 70%-80%, the ethanol water of preferably 75%.The present inventor is found through experiments, with
The ethanol water of 70%-80% extracts the epicarpium Castaneae extract obtained and has more preferably suppression PTP1B activity.
Wherein in an embodiment, in described extraction step, described raw materials quality and the ratio of solvent volume
For 1g:5-10ml, divide and carry out ultrasonic extraction, each 30min-60min for 2-4 time.Method described above is extracted,
There is extraction efficiency high, strong operability, obtain the advantage that its lytic activity is good.
Wherein in an embodiment, also include that purification step, described purification step are:
The epicarpium Castaneae extract loading obtained by described extraction step, to macroporous adsorbent resin, is purified, uses body
The long-pending ratio methanol-water solution eluting for 1-3:8 (preferably 2:8), collects eluent, recycling design, to obtain final product.
By further purification, it is possible to be enriched with in the compositions of this extract and really have PTP1B suppression to live
The composition of property.Further, the present inventor finds after substantial amounts of experiment, has the one-tenth of inhibitory activity to PTP1B
Dividing in the methanol-water eluent being primarily present in 1-3:8, in the eluent of other component, the activity of composition is the most not
Such as this component.
Wherein in an embodiment, in described purification step, the model of described macroporous adsorbent resin is HP-20
Type.With the purification with macroreticular resin of this model above-mentioned epicarpium Castaneae extract, it is possible to be preferably enriched with this extract
Compositions in the real composition that PTP1B is had inhibitory activity.
Wherein in an embodiment, in described purification step, the epicarpium Castaneae obtained by described extraction step extracts
Thing loading to after macroporous adsorbent resin, is the methanol-water solution of 0:10,0.5-2:9,1-3:8 by volume ratio successively
Gradient elution, collected volume, than the eluent of the methanol-water solution for 1-3:8 part, recycling design, to obtain final product.
First with water and methanol-water solution gradient elution that volume ratio is 0.5-2:9, remove wherein partial impurities composition, tool
There is preferable purification effect.
Wherein in an embodiment, in described purification step, the methanol-water solution with volume ratio as 0:10 is washed
De-1-3 column volume；1-3 column volume of methanol-water solution eluting with volume ratio as 0.5-2:9；With body
Long-pending than 1-3 the column volume of methanol-water solution eluting for 1-3:8.With the elution of above-mentioned column volume, energy
Enough removal impurity components to greatest extent also retain the composition having inhibitory activity to PTP1B.
The epicarpium Castaneae extract that the bright preparation method also disclosing above-mentioned epicarpium Castaneae extract of this law prepares.Should
Epicarpium Castaneae extract has the effect of PTP1B inhibitory activity, for a kind of natural PTP1B inhibitor, and can be as pressing down
The inhibitor of protein-tyrosine-phosphatase 1B processed and/or use as euglycemic agent, for prevention and
Treatment diabetes, obesity and complication thereof.
This law is bright also discloses the suppression as suppression protein-tyrosine-phosphatase 1B of the above-mentioned epicarpium Castaneae extract
Agent and/or as the application in euglycemic agent.
Wherein in an embodiment, the inhibitor of described protein-tyrosine-phosphatase 1B, and/or described
Euglycemic agent is applied in the medicine preventing and treating diabetes, obesity and complication thereof in preparation.
Compared with prior art, the method have the advantages that
The preparation method of a kind of epicarpium Castaneae extract of the present invention, uses the epicarpium Castaneae extract that the method prepares,
There is the effect of suppression protein-tyrosine-phosphatase 1B (PTP1B), it is possible to using this extract as PTP1B
Inhibitor uses, for treating diabetes, obesity and other complication thus caused.To exploitation
The resources of medicinal plant of China is significant.
Further, the preparation method of this epicarpium Castaneae extract has strong operability, it is adaptable to the advantage of industrialized production.
Accompanying drawing explanation
Fig. 1 is the impact that in embodiment, extract causes hyperglycemic rat insulin sensitivity respectively to STZ；
Fig. 2 is the impact that in embodiment, extract causes hyperglycemic rat carbohydrate tolerance respectively to STZ.
Detailed description of the invention
For the ease of understanding the present invention, below with reference to relevant drawings, the present invention is described more fully.
Accompanying drawing gives presently preferred embodiments of the present invention.But, the present invention can come real in many different forms
Existing, however it is not limited to embodiment described herein.On the contrary, providing the purpose of these embodiments is to make this
The understanding of disclosure of the invention content is more thorough comprehensively.
Unless otherwise defined, all of technology used herein and scientific terminology and the technology belonging to the present invention
The implication that the technical staff in field is generally understood that is identical.The art used the most in the description of the invention
Language is intended merely to describe the purpose of specific embodiment, it is not intended that in limiting the present invention.Used herein
Term "and/or" includes the arbitrary and all of combination of one or more relevant Listed Items.
In following example, macroporous resin (HP-20) chromatograph used, for Beijing green BAICAO development in science and technology
Company limited；STZ (streptozotocin), sanland product, analytical pure, use in 4 DEG C of ice baths before use
0.05mol/ml citric acid pH 4.5 solution is prepared, and uses immediately；Diabetes pill, the limited public affairs of Pharmaceutical in Guangzhou
Department, lot number: GF0085.
Embodiment 1
A kind of epicarpium Castaneae extract, is prepared by the following method and obtains:
1, extract.
By Semen Castaneae (the Castanea mollissima Blume) shell of dry chopping, it is 100% by concentration expressed in percentage by volume
The ethanol (i.e. straight alcohol) of concentration, is 5-10 times amount according to chestnut shell and ethanol solid-liquid ratio (g/ml), carries out
Ultrasonic extraction 2-4 time, each 30min-60min, united extraction liquid.In the present embodiment, chestnut shell is
500g, ethanol is 5L, ultrasonic extraction 3 times, each 60min.
Subsequently, extracting solution concentrating under reduced pressure reclaims ethanol, to more difficult concentration, merges concentrated solution, continues at uncovered
Vessel in heating waves most ethanol as far as possible, cools, and obtains 100% ethanol extract 32g, obtains epicarpium Castaneae after drying
Extract, is component I.
2, purification.
The epicarpium Castaneae extract loading obtained by described extraction step, to macroporous adsorbent resin (HP-20), carries out pure
Changing, using methanol/water (v/v, 0:10,1:9,2:8,10:0) respectively is eluent, every kind of eluent 1L (phase
When in 2 column volumes), carry out gradient elution, collect the separation component of methanol/water (v/v, 2:8), collect
Eluent (methanol/water=2:8), by eluent room temperature concentrating under reduced pressure, is dried, obtains epicarpium Castaneae extract and refine component
4.3g, is component H I.
Embodiment 2
A kind of epicarpium Castaneae extract, is prepared by the following method and obtains:
1, extract.
By Semen Castaneae (the Castanea mollissima Blume) shell of dry chopping, it is 75% by concentration expressed in percentage by volume
The ethanol water of concentration, is 5-10 times amount according to chestnut shell and ethanol solid-liquid ratio (g/ml), carries out ultrasonic
Ripple extracts 2-4 time, each 30min-60min, united extraction liquid.In the present embodiment, chestnut shell is 500g,
Ethanol is 5L, ultrasonic extraction 3 times, each 60min.
Subsequently, extracting solution concentrating under reduced pressure reclaims ethanol, to more difficult concentration, merges concentrated solution, continues at uncovered
Vessel in heating waves most ethanol as far as possible, cools, and obtains 100% ethanol extract 38.2g, obtains chestnut after drying
Shell extract, is component II.
2, purification.
The epicarpium Castaneae extract loading obtained by described extraction step, to macroporous adsorbent resin (HP-20), carries out pure
Changing, be eluent by methanol/water (v/v, 0:10,1:9,2:8,10:0), every kind of eluent 1L is (quite
In 2 column volumes), carry out gradient elution, collect the separation component of methanol/water (v/v, 2:8), collection is washed
De-liquid (methanol/water=2:8), by eluent room temperature concentrating under reduced pressure, is dried, obtains epicarpium Castaneae extract and refine component 6.2
G, is component H II.
Further, in this embodiment, also by normal for the eluent of alcohol/water (v/v, 0:10,1:9,10:0) component
Temperature concentrating under reduced pressure, is dried, respectively obtains component A, component B and component C.
Embodiment 3
A kind of epicarpium Castaneae extract, is prepared by the following method and obtains:
1, extract.
By Semen Castaneae (the Castanea mollissima Blume) shell of dry chopping, it is 50% by concentration expressed in percentage by volume
The ethanol water of concentration, is 5-10 times amount according to chestnut shell and ethanol solid-liquid ratio (g/ml), carries out ultrasonic
Ripple extracts 2-4 time, each 30min-60min, united extraction liquid.In the present embodiment, chestnut shell is 500g,
Ethanol is 5L, ultrasonic extraction 3 times, each 60min.
Subsequently, extracting solution concentrating under reduced pressure reclaims ethanol, to more difficult concentration, merges concentrated solution, continues at uncovered
Vessel in heating waves most ethanol as far as possible, cools, and obtains 100% ethanol extract 28.5g, obtains chestnut after drying
Shell extract, is component III.
2, purification.
The epicarpium Castaneae extract loading obtained by described extraction step, to macroporous adsorbent resin (HP-20), carries out pure
Changing, be eluent by methanol/water (v/v, 0:10,1:9,2:8,10:0), every kind of eluent 1L is (quite
In 2 column volumes), carry out gradient elution, collect the separation component of methanol/water (v/v, 2:8), collection is washed
De-liquid (methanol/water=2:8), by eluent room temperature concentrating under reduced pressure, is dried, obtains epicarpium Castaneae extract and refine component 3.1g,
It is component H III.
Test example 1
The suppression PTP1B activity test of the extract that above-described embodiment obtains.
1, experimental technique.
It is the GST from expression in escherichia coli purification for the protein-tyrosine-phosphatase PTP1B of screening
Fusion protein (source: the U.S., article No.: A0230).Use ultraviolet substrate pNPP, observe variable concentrations counterweight
The activity suppression of group enzyme, with the medicinal effects of preliminary assessment compound.
The product obtained due to the phospholipid of PTP1B hydrolysis substrate pNPP has the strongest light to absorb at 410nm.
Therefore change that light at 410nm absorbs can directly be detected to observe the activity change of enzyme, and compound pair
Its suppression situation.And with sodium vanadate as positive control, its IC50It is 2.0 μm ol/L.Experimental result is as follows
Shown in table:
The different extract IC to PTP1B inhibitory activity of table 1.50(n=10)
Experimental result shows: different concentration ethanol extract and active constituent are to suppression PTP1B active effect all
Differ, wherein component II and component H II (75% ethanol extraction and 75% ethanol extraction refine component)
Optimal to suppression PTP1B active effect, its IC50It is respectively 15.5 ± 1.5 μ g/ml and 11.3 ± 1.4 μ g/ml.
And component A, B, C are compared with component H II, almost without activity, illustrate in 75% ethanol extraction,
The active component with suppression PTP1B effect is primarily present in the eluent of methanol/water (v/v, 2:8).
Test example 2
The extract that above-described embodiment obtains causes the experiment of hyperglycemic rat blood sugar lowering respectively to STZ.
1, experimental technique.
With Kunming rat 100 (male), random packet become blank group, model group (Diabetic control),
Experimental group (component I, component H I, component II, component H II, component III, component H III), diabetes pill
Matched group (XKW).
In addition to blank group, water 1d is can't help in the fasting of each group, (uses 0.05mol/ml in 4 DEG C of ice baths with STZ
Citric acid pH4.5 solution is prepared, and uses immediately) lumbar injection 60mg/kg, makes diabetes model.Injection
Rear 72h docking measuring blood sugar of blood extracting (fasting 6h before surveying), blood glucose value is used for testing higher than 11.1mmol/L
(10/group).
Experimental group is respectively with component I, component H I, component II, component H II, component III, component H III
Gavage (200mg/kg)；XKW group is with diabetes pill gavage (250mg/kg)；Blank group, model group gavage
Equal-volume normal saline.It is administered 30d, weighs every day, be administered once.After last is administered, docking takes blood and surveys
Determine blood glucose (fasting 6h before surveying).Experimental result is as follows:
The different extract of table 2. causes hyperglycemic rat blood sugar lowering situation to STZ
Experimental result shows: experimental group rat blood sugar all has decline in various degree, wherein with component II and group
The experimental group of part H II (75% ethanol extraction and 75% ethanol extraction refine component) gavage and model group ratio
Relatively, blood glucose effect is reduced the most notable.Experimental data shows, the hypoglycemic effect of component H II is suitable with diabetes pill.
Test example 3
The extract that above-described embodiment obtains causes the impact experiment of hyperglycemic rat insulin sensitivity to STZ.
1, experimental technique.
Experimental group is respectively with component I, component H I, component II, component H II, component III, and component H III fills
Stomach (200mg/kg)；XKW group is with diabetes pill gavage (250mg/kg)；Blank group, model group gavage etc.
ML normal saline.It is administered 30d, weighs every day, be administered once.The most each group being administered in 30d, often
It lumbar injection Depot H Insulin 1IU/kg.After 30d, each group rat every day of lumbar injection respectively in four days
Rapid acting human insulin 0.05,0.5,1.0,2.5IU/kg, measure to the blood glucose before and after rapid acting human insulin, meter
Calculate ratio.Experimental result is as shown in Figure 1.
Showing from the experimental result of Fig. 1, different concentration ethanol extract and active constituent cause high blood to STZ
Sugar rat insulin sensitivity all has improvement, but each component effect is different.Wherein component II and component H II (75%
Ethanol extraction and 75% ethanol extraction refine component) compare with model group, insulin sensitivity strengthens effect
Fruit is preferable, component H II most pronounced effects.Experimental data shows, the hypoglycemic effect of component H II and XZK group
Quite.
Test example 4
The extract that above-described embodiment obtains causes the impact experiment of hyperglycemic rat carbohydrate tolerance respectively to STZ.
1, experimental technique
Experimental group is respectively with component I, component H I, component II, component H II, component III, and component H III fills
Stomach (300mg/kg)；XKW group is with diabetes pill gavage (250mg/kg)；Blank group, model group gavage etc.
ML normal saline (often group 10).It is administered 30d, weighs every day, be administered once.Below administration is respectively organized
Fasting 4h (9:00～13:00) after 30d, gavage glucose (300mg/kg), measure before gavage (0min)
And 30min after gavage, 60min, 120min blood glucose.Experimental result is as shown in Figure 2.
Showing from the experimental result of Fig. 1, each group medicine STZ that is improved causes the carbohydrate tolerance of hyperglycemic rat
Trend, but each component effect is different.Wherein component II and component H II (75% ethanol extraction and 75%
Ethanol extraction refines component) effect is preferable, component H II best results.Experimental data shows, component H
The hypoglycemic effect of II is better than XZK group.
Test example 5
The extract that above-described embodiment obtains causes the experiment of hyperglycemic rat body weight change respectively to STZ.
Experimental group is respectively with component I, component H I, component II, component H II, component III, and component H III fills
Stomach (300mg/kg)；XKW group is with diabetes pill gavage (250mg/kg)；Blank group, model group gavage etc.
ML normal saline.It is administered 30d, weighs every day, be administered once.Experimental result is as follows:
The different extract of table 3. causes hyperglycemic rat body weight change situation (n=10) to STZ
Experimental result shows: different concentration ethanol extract and active constituent cause hyperglycemic rat body weight to STZ
Reduction effect all differ.Wherein component II and component H II (75% ethanol extraction and 75% ethanol extraction
Thing refines component) compare with model group, reduce body weight best results.Experimental data shows, component H II
Hypoglycemic effect is suitable with XKW group.
By the experimental result of above-mentioned test example, think: insulin sensitivity reduces and beta Cell of islet
Function is impaired is the pathogenetic principal element of type 2 diabetes mellitus, and the height of PTP1B is expressed and can be caused body
Insulin sensitivity is reduced and leptin resistance, thus causes type 2 diabetes mellitus and obesity.
Test data shows, the epicarpium Castaneae extract of the present invention can significantly inhibit PTP1B activity (test example 1).
And cause hyperglycemic rat blood sugar lowering experiment (test example 2) by STZ, affirm fully extract pair of the present invention
The hypoglycemic activity of diabetic animal models, also points out each group of medicine all to have diabetes-alleviating clinic polydipsia, many
The action character of urine symptom, and show (examination by STZ cause hyperglycemic rat insulin sensitivity Journal of Sex Research further
Test example 3), extract of the present invention can be obviously improved the body sensitivity to insulin, improves the life of insulin
Thing availability.By STZ is caused hyperglycemic rat carbohydrate tolerance detection display (test example 4), the chestnut of the present invention
Shell extract can significantly improve body carbohydrate tolerance.(test example is learnt by rat body weight change test data
5), extract of the present invention all has remarkable result to suppression body weight increase.
Summary correlation test data, show extract of the present invention to treatment type 2 diabetes mellitus, obesity and
Complication has potential practical significance.
Each technical characteristic of embodiment described above can combine arbitrarily, for making description succinct, the most right
The all possible combination of each technical characteristic in above-described embodiment is all described, but, if these skills
There is not contradiction in the combination of art feature, is all considered to be the scope that this specification is recorded.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed,
But can not therefore be construed as limiting the scope of the patent.It should be pointed out that, for this area
For those of ordinary skill, without departing from the inventive concept of the premise, it is also possible to make some deformation and change
Entering, these broadly fall into protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be with appended power
Profit requires to be as the criterion.
Claims (10)
1. the preparation method of an epicarpium Castaneae extract, it is characterised in that comprise the following steps:
Extract: with chestnut shell as raw material, the ethanol water with concentration expressed in percentage by volume as 40%-100% as solvent,
Carry out solvent extraction, recycling design, obtain epicarpium Castaneae extract.
The preparation method of epicarpium Castaneae extract the most according to claim 1, it is characterised in that described extraction
In step, described solvent be concentration expressed in percentage by volume be the ethanol water of 70%-80%.
The preparation method of epicarpium Castaneae extract the most according to claim 1, it is characterised in that described extraction
In step, the ratio of described raw materials quality and solvent volume is 1g:5-10ml, point carries out ultrasound wave for 2-4 time and carries
Take, each 30min-60min.
4. according to the preparation method of the epicarpium Castaneae extract described in any one of claim 1-3, it is characterised in that
Also include that purification step, described purification step are:
The epicarpium Castaneae extract loading obtained by described extraction step, to macroporous adsorbent resin, is purified, uses body
The long-pending ratio methanol-water solution eluting for 1-3:8, collects eluent, recycling design, to obtain final product.
The preparation method of epicarpium Castaneae extract the most according to claim 4, it is characterised in that described purification
In step, the model of described macroporous adsorbent resin is HP-20 type.
The preparation method of epicarpium Castaneae extract the most according to claim 4, it is characterised in that described purification
In step, after the epicarpium Castaneae extract loading obtained by described extraction step to macroporous adsorbent resin, use body successively
Long-pending than being the methanol-water solution gradient elution of 0:10,0.5-2:9,1-3:8, collected volume ratio is for 1-3:8 part
The eluent of methanol-water solution, recycling design, to obtain final product.
The preparation method of epicarpium Castaneae extract the most according to claim 6, it is characterised in that described purification
1-3 column volume of methanol-water solution eluting in step, with volume ratio as 0:10；With volume ratio as 0.5-2:
1-3 column volume of methanol-water solution eluting of 9；Methanol-water solution eluting 1-3 with volume ratio as 1-3:8
Column volume.
8. the epicarpium Castaneae that the preparation method of the epicarpium Castaneae extract described in any one of claim 1-7 prepares extracts
Thing.
9. the epicarpium Castaneae extract described in claim 8 is as the suppression of suppression protein-tyrosine-phosphatase 1B
Agent and/or as the application in euglycemic agent.
Application the most according to claim 9, it is characterised in that: described protein-tyrosine-phosphatase
The inhibitor of 1B, and/or described euglycemic agent preparation be used for preventing and treat diabetes, obesity and
The medicine of its complication is applied.
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| CN103948654A (en) * | 2014-05-13 | 2014-07-30 | 北京林业大学 | Method of extracting, purifying and inhibiting alpha-glucosaccharase active ingredient from chestnut shell |
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| CN103948654A (en) * | 2014-05-13 | 2014-07-30 | 北京林业大学 | Method of extracting, purifying and inhibiting alpha-glucosaccharase active ingredient from chestnut shell |
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| CN108523126A (en) * | 2018-04-24 | 2018-09-14 | 广州正广生物科技有限公司 | It is a kind of that there is the composition and preparation method thereof for improving blood glucose level |
| CN108523126B (en) * | 2018-04-24 | 2021-11-09 | 广州正广生物科技有限公司 | Composition for improving blood sugar level and preparation method thereof |
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