CN106265717A - Dicliptera chinensis polysaccharide application in preparation preventing and treating diabetes medicament or health product - Google Patents
Dicliptera chinensis polysaccharide application in preparation preventing and treating diabetes medicament or health product Download PDFInfo
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- CN106265717A CN106265717A CN201610576684.9A CN201610576684A CN106265717A CN 106265717 A CN106265717 A CN 106265717A CN 201610576684 A CN201610576684 A CN 201610576684A CN 106265717 A CN106265717 A CN 106265717A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- 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)
- A61K36/19—Acanthaceae (Acanthus family)
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Abstract
The application in preparation preventing and treating diabetes medicament or health product of the present invention open dicliptera chinensis polysaccharide.Test by preparing diabetic animal, find that it can improve the symptom of diabetes rat, suppression death, maintain body weight, reduce blood glucose and improve resistance to sugar amount, showing its good blood sugar reducing function.In addition, dicliptera chinensis polysaccharide can reduce diabetes rat serum advanced glycation end products peptide (AGE P), glycolated hemoglobin (HbAlc), T-CHOL (TC), triglyceride (TG), low density lipoprotein, LDL (LDL C) and kidney aldose reductase (AR) activity or content, raise liver glycogen and high density lipoprotein (HDL C) content, illustrate that it by suppression liver, nephropathy and improves blood fat environment, reach to suppress the effect of diabetic complication.Be experimentally confirmed dicliptera chinensis polysaccharide have preferably prevent and treat diabetes and suppression complication effect, this new application is that its medical value has opened up frontier, has important using value and economic benefit.
Description
Technical field
The invention belongs to tcm field, relate to Herba Diclipterae Chinensis Dicliptera chinensis (L.) Nees. herb be
Raw material extraction purification goes out dicliptera chinensis polysaccharide and the application in being prepared as preventing and treating diabetes medicament or health product thereof,
Background technology
Herba Diclipterae Chinensis is that Acanthaceae China Herba Peristrophes belongs to the complete of Herba Diclipterae Chinensis Dicliptera chinensis (L.) Nees.
Grass.Herb sweet in the mouth, micro-hardship, cold in nature, GUIXIN, liver, lung meridian, have heat clearing away, removing heat from blood, dampness removing, detoxify, promote the production of body fluid, the effect such as diuresis, face
Bed be usually used in treating cold, fever, calentura send out speckle, summer-heat excessive thirst, tell epistaxis, have blood in stool, hematuria, metrorrhagia, cough due to lung-heat, swelling throat
Bitterly, the card such as liver-heat conjunctival congestion, eye conjunctivitis, infantile convulsion, dribbling urination, leukorrhagia, herpes zoster, carbuncle furuncle furuncle, Serpentis lyssodexis.Canis familiaris L.
Liver dish resource is the abundantest, is distributed the ground such as Yunnan, Guangdong, Guangxi, for southwestern medicinal herbs most in use among the people.Herba Diclipterae Chinensis is Guangxi Folk medicine
Eat dual-purpose conventional strong medicine, mainly contain the materials such as polysaccharide, organic acid, aminoacid.Invention person's early-stage Study finds Canis familiaris L.
Liver dish polysaccharide component has antiinflammatory, antioxidation and raising immunity of organism, reaches to improve and regulation organism metabolism effect, but antagonism sugar
The research of urine disease still belongs to the first time.
Diabetes (Diabetes mellitus, DM) are often referred to type 2 diabetes mellitus (TZDM), account for more than the 90% of DM sum,
Its sickness rate, dead rate are high, worldwide have become as the third-largest serious harm after cardiovascular and cerebrovascular disease and cancer
The disease of human health.Diabetes are jointly to be acted on by heredity and acquired environment, chronic based on carbohydrate metabolism disturbance
Comprehensive disease.Along with disease, can cause organizing multiple organ pathological changes, serious threat human health more.At traditional sugar
In urine disease treatment, diabetes treatment by western medicine instant effect, but need long-term taking, and erious adverse reaction.Modern study finds, in part
Medicine or its extract, in terms for the treatment of diabetes, have stable curative effect and untoward reaction are few, have the irreplaceable feature of Western medicine
And advantage.Therefore from natural drug, find safe and effective antidiabetic medicine tool be of great significance.Herba Diclipterae Chinensis medicine
With aboundresources, it is thus achieved that the low cost of dicliptera chinensis polysaccharide, suitable a large amount of productions.Retrieve the research of domestic and international dicliptera chinensis polysaccharide, still
Have no research report dicliptera chinensis polysaccharide prevented and treated in terms of diabetes effect.
Summary of the invention
It is an object of the invention to go out dicliptera chinensis polysaccharide with Herba Diclipterae Chinensis herb for raw material extraction purification, and be prepared into preventing and treating
Diabetes medicament or health product, this announcement that still belongs to the first time at home.
The technical scheme realizing the object of the invention is:
The new application of dicliptera chinensis polysaccharide, prevents and treats medicine or the health product of diabetes for preparation, can be prepared into for mouth
The Chinese medicine preparation of clothes, such as powder, tablet, oral liquid, granule, capsule and pill etc., its preparation method and existing corresponding dosage form
Preparation method is general.
The preparation of dicliptera chinensis polysaccharide of the present invention is to weigh dry Herba Diclipterae Chinensis herb coarse powder, adds petroleum ether backflow defat and carries
Take 1-2h, after medicinal residues volatilization petroleum ether to the greatest extent, boiling water extraction 1-3 time, each 1-3h, merge and concentrated extracting solution, with chloroform and just
Ratio 4:1 of butanol, uses Sevag fado time extraction, the supernatant of the protein that is removed, adds dehydrated alcohol and make alcohol content reach
To 70-99%, precipitate with ethanol in 4 DEG C of refrigerators, sucking filtration, abandoning supernatant, filtering residue uses ether, dehydrated alcohol, washing with acetone, i.e. successively
Obtain Herba Diclipterae Chinensis crude polysaccharides；Herba Diclipterae Chinensis crude polysaccharides is dissolved in water, is loaded onto the DE52 of phosphate buffer pre-treatment through pH=7-9
In type resin chromatography post, static 12h, with distilled water, flow velocity is with 1-3mL/min eluting, then through sephadex-G25 gel chromatography
Post, with distilled water, flow velocity is with 0.5-1mL/min eluting, concentrate eluant, then removes its polysaccharide pigment through activated carbon, and vacuum is done
Dry white powder dicliptera chinensis polysaccharide (DCP).
Compared with prior art, Herba Diclipterae Chinensis herb resource enriches the present invention, prepares that dicliptera chinensis polysaccharide technique is simple, cost
Low, without any side effects, in good taste, there is good Development volue.It is applicable to additionally, the present invention has excavated dicliptera chinensis polysaccharide
Prevent and treat diabetes, determined curative effect, at preparation preventing and treating diabetes medicament or health product, there is good medical value, and be Herba Diclipterae Chinensis
Polysaccharide deeply develops offer Research foundation further.
Accompanying drawing explanation
Fig. 1 is the impact (n=10) on diabetes rat body weight of the embodiment of the present invention dicliptera chinensis polysaccharide；
Fig. 2 is the impact on blood glucose in diabetic rats concentration of the embodiment of the present invention dicliptera chinensis polysaccharide；
Fig. 3 is the impact on diabetes rat carbohydrate tolerance of the embodiment of the present invention dicliptera chinensis polysaccharide；
In figure: A, B, C, D, E, F correspondence respectively represents blank group, model group, the positive (glibenclamide) group, Herba Diclipterae Chinensis
Polysaccharide high, medium and low dosage group,
In the abscissa of Fig. 1 and Fig. 2, before "-1 " refers to modeling；" 0 " refers to modeling 72h；" 1,2,3,4,5,6 " show medicine week number,
In the abscissa of Fig. 3,0,0.5,1,1.5,2.0,2.5,3.0 refer to minute (h).
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, present invention is further described, but is not limitation of the invention.
One, the following is the extraction and purification method of dicliptera chinensis polysaccharide:
Weigh dry Herba Diclipterae Chinensis herb coarse powder 2000g, add the backflow defat of 20000ML petroleum ether and extract 2h.Medicinal residues are waved
To the greatest extent after petroleum ether, boiling water extraction 2 times, each 1.5h, united extraction liquid, it is concentrated into 200mL, with chloroform-n-butyl alcohol (4:1) repeatedly
Extraction (i.e. Sevag method), the supernatant of the protein that is removed, add 95% ethanol and make alcohol content reach 80%, in 4 DEG C of refrigerators
Precipitate with ethanol 12h.Sucking filtration, abandoning supernatant, filtering residue with ether, dehydrated alcohol, washing with acetone, obtains Herba Diclipterae Chinensis crude polysaccharides successively.Will
Herba Diclipterae Chinensis crude polysaccharides is dissolved in water, is loaded onto in the DE52 type resin chromatography post of the phosphate buffer pre-treatment of PH=8,
Static 12h, with distilled water, flow velocity is with 2mL/min eluting, then through sephadex-G25 gel chromatography column, with distilled water, flow velocity with
0.5-1mL/min eluting, concentrate eluant, then remove its polysaccharide pigment through activated carbon, it is vacuum dried to obtain white powder Canis famililiaris L.
Dish polysaccharide (DCP).Dicliptera chinensis polysaccharide glucose compares product, uses phenol one sulfuric acid method, through UV-vis spectroscopy meter
The purity of polysaccharide measured in sample reaches 95.6%.
Two, dicliptera chinensis polysaccharide is prepared as pharmaceutical preparation:
Dicliptera chinensis polysaccharide can be added corresponding adjuvant, be prepared as powder, tablet, oral liquid, granule, capsule and pill
Deng.
Three, dicliptera chinensis polysaccharide to diabetes rat blood sugar lowering and suppresses complication effect experiment method and result:
1, material, reagent and instrument
1.1 animal
SPF level SD rat 90, male and female half and half, Medical Colleges Of Guilin's SPF Experimental Animal Center provide (credit number
SCXK2007-0001)。
1.2 medicines and reagent
Herba Diclipterae Chinensis is accredited as Acanthaceae China Herba Peristrophes through Guangxi University of Chinese Medicine professor Wei Songji and belongs to Herba Diclipterae Chinensis
The herb of Dicliptera chinensis (L.) Nees..Reality is prepared with " the extraction and purification method of dicliptera chinensis polysaccharide "
Test with dicliptera chinensis polysaccharide (DCP) for reagent product.
Glibenclamide, Huarun Zizhu Pharmaceutical Co., Ltd..Streptozotocin (STZ), is purchased from Sigma company.T-CHOL
(TC), triglyceride (TG), high density lipoprotein (HDL-C), low density lipoprotein, LDL (LDL-C), be purchased from Nanjing and build up biological work
Journey institute.Insulin radioimmunoassay immune reagent kit, is purchased from Bio-Rad company.Advanced glycation end products one peptide (AGE-P), liver glycogen,
Kidney aldose reductase (AR) and glycolated hemoglobin (HbAlc) detection kit, be purchased from the Zhejiang limited public affairs of Ai Kang biotechnology
Department.
1.3 instrument
TLL-C tabletop refrigerated centrifuge, Fourth Ring, Beijing scientific instrument factory；TSN-695B type is put and is exempted from measuring instrument, Shanghai day ring
Instrument one factory；AU600 full-automatic biochemical detector, Olympus produces；752 type spectrophotometers, Shanghai the 3rd analytical tool
Factory；BS1105 electronic balance, sartorius company of Germany；Bayer blood glucose monitoring system and reagent paper thereof, Germany Bayer.
2. experimental technique
90 SD rats are randomly divided into Normal group (10) and modeling group (80), and Normal group is fed commonly to raise
Material, modeling group is fed with high glucose and high fat feedstuff 4 weeks.After 4 weeks, all Rat Fast can't help water 12h, and modeling group presses 35mg/kg abdominal cavity note
Penetrating streptozotocin (STZ) modeling, Normal group only injects equivalent lemon acid-sodium citrate buffer solution (pH=4.4), after injection
Continue fasting and can't help water 12h.After again feeding 72h, water 12h is can't help in fasting, measures the fasting glucose of modeling group, sieve by blood glucose meter
Selecting fasting glucose >=13.8mmol/L is the successful diabetes rat of modeling, and this tests into mould 74.By successful for modeling rat
It is divided into model group, positive controls (glibenclamide, 20mg/kg) and dicliptera chinensis polysaccharide according to blood glucose value and body weight stochastic averagina
High, medium and low dosage group (600,400,200mg/kg), gastric infusion, Normal group and model group gavage distilled water, fill continuously
Stomach totally 6 weeks.Observe the general status of animal during experiment, weigh in each week 1 time before being administered and after being administered, and quiet through animal tail
Arteries and veins is taken a blood sample, and measures whole blood blood glucose.After being administered 6 weeks, water 12h is can't help in fasting, and each group rat gavage respectively gives glucose 2.5g/
Kg, measures blood glucose value respectively at 0h, 0.5h, 1h, 2h, 3h tail venous blood sampling, draws change of blood sugar figure, and t checks the more individual time
The glucose-tolerant degree of point.After water 4h is can't help in fasting again, chloral hydrate anesthesia, abdominal aortic blood, take liver and kidney.
3. experimental result
The observation of 3.1 rat general status
The normal rats mental status is good, and body weight increases, and moves freely.Diabetic model group rat occurs the most
Food, polydipsia, polyuria, lose weight symptom, and tired mind, movable reduces, and hair is lackluster, 4 death of experiment period；Canis familiaris L.
Liver dish polysaccharide group rat amount of drinking water, urine volume, dietary amount the most relatively model group substantially reduce, and body weight relatively diabetes rat dramatically increases,
There is 1 death in only low dosage, illustrates that dicliptera chinensis polysaccharide can improve the symptom of diabetes rat.
The change of 3.2 rat body weights
After rats by intraperitoneal injection STZ 72h, each modeling group and modeling precursor heavy phase ratio, all it is remarkably decreased (P ＜ 0.01).Real
Testing period, blank group body weight is gradually increased, modeling group rat all in reducing trend, wherein model group rats body weight the most all under
Decline particularly evident in fall trend, the especially the last fortnight after modeling, and medication group pharmaceutical intervention is after two weeks, positive group and Canis famililiaris L.
Dish polysaccharide high dose group body weight from the 2nd week is the most in rising trend, in dicliptera chinensis polysaccharide, low dose group body weight keep stable.
See Fig. 1.
The change of 3.3 rat blood sugar levels
Before administration, compare with blank group blood sugar level, modeling group statistically significant (P ＜ 0.01), illustrate that modeling becomes
Merit.After administration, from the 2nd week, comparing with model group, dicliptera chinensis polysaccharide each dosage group and positive group blood glucose substantially reduce, and all have
Statistical significance (P ＜ 0.05).See Fig. 2.
3.4 oral glucose tolerances (OGTT) are tested
OGTT experiment is the goldstandard diagnosing DM clinically.Experimental result shows, compared with model group, and positive group and Canis famililiaris L.
Dish polysaccharide high, medium and low dosage group can significantly improve diabetes rat glucose tolerance, the blood when gavage gives glucose 3h
Sugar value difference heteropole is notable.See Fig. 3.
3.5 dicliptera chinensis polysaccharides are to diabetes rat serum advanced glycosylation end products-peptide (AGE-P) and glycolated hemoglobin
(HbAlc) impact
Monitoring advanced glycosylation end products-peptide (AGE-P) arranges for the examination of DM high-risk group, glycolated hemoglobin (HbAlc)
One of main standard for DM and the diagnosis of its complication.Experimental result shows, compared with model group, medication group rat AGE-P and
HbAlc all has significance to reduce, and shows that dicliptera chinensis polysaccharide can improve diabetes and its complication, this result table 1 and Fig. 2 result
Consistent.
The table 1 dicliptera chinensis polysaccharide (DCP) impact on diabetes rat AGE-P and HbAlc
Compare with model group, * P ＜ 0.05, * * P ＜ 0.01 (following table represents same with dosage).
3.6 dicliptera chinensis polysaccharides are to diabetes rat hepatic tissue liver glycogen, kidney aldose reductase (AR) activity and level
Impact
The effect of liver glycogen is the most stable of maintenance blood sugar concentration, and (such as diabetes) under hyperglycemic state, hexose swashs
Enzyme is saturated, at this moment AR enzyme activition, promotes internal nephropathy.Experimental result shows, compared with model group, and medication
Group rats'liver glycogen raises and kidney AR significance reduces, and shows that dicliptera chinensis polysaccharide can regulate and control diabetes glucose concentration and suppression
The complication of kidney.It is shown in Table 2.
Table 2 dicliptera chinensis polysaccharide (DCP) is on diabetes rat liver glycogen and the impact of A R
3.7 dicliptera chinensis polysaccharide is on blood fat factor TC, the impact of TG, HDL-C and LDL-C level in diabetes rat serum
Compared with blank group, the equal significance of model group rats TC, TG, LDL-C level raises, and the horizontal significance of HDL-C drops
Low, show that serious diseases occurs in the blood lipid metabolism of diabetes rat.Through the pharmaceutical intervention of 6 weeks, compared with model group, Herba Diclipterae Chinensis
Polysaccharide each dosage group rat equal significance of TC, TG, LDL-C level reduces, and the horizontal significance of HDL-C raises, and shows that Herba Diclipterae Chinensis is many
Sugar can have improvement result to the blood fat environment of diabetes rat.It is shown in Table 3.
Table 3 dicliptera chinensis polysaccharide (DCP) is to the impact of LDL-C, HDL-C, TG and TC in diabetes rat serum
By above experiment, find that dicliptera chinensis polysaccharide can improve the symptom of diabetes rat, suppression death, maintain body
Weight, reduces blood glucose and improves resistance to sugar amount, showing its good blood sugar reducing function.Additionally, dicliptera chinensis polysaccharide can to reduce diabetes big
Mus serum advanced glycosylation end products-peptide (AGE-P), glycolated hemoglobin (HbAlc), T-CHOL (TC), triglyceride (TG), low
Density lipoprotein (LDL-C) and kidney aldose reductase (AR) activity or content, raise liver glycogen and high density lipoprotein (HDL-
C) content, illustrates that it by suppression liver, nephropathy and improves blood fat environment, reaches to suppress the effect of diabetic complication.
Claims (2)
1. dicliptera chinensis polysaccharide application in preparation preventing and treating diabetes medicament or health product.
Application the most according to claim 1, is characterized in that: the preparation of described dicliptera chinensis polysaccharide is to weigh dry Herba Diclipterae Chinensis
Herb coarse powder, adds petroleum ether backflow defat and extracts 1-2h, after medicinal residues volatilization petroleum ether to the greatest extent, and boiling water extraction 1-3 time, each 1-
3h, merges and concentrated extracting solution, by ratio 4:1 of chloroform Yu n-butyl alcohol, uses Sevag fado time extraction, and be removed albumen
The supernatant of matter, adds dehydrated alcohol and makes alcohol content reach 70-99%, precipitate with ethanol in 4 DEG C of refrigerators, sucking filtration, abandoning supernatant, filtering residue
Successively with ether, dehydrated alcohol, washing with acetone, obtain Herba Diclipterae Chinensis crude polysaccharides；Herba Diclipterae Chinensis crude polysaccharides is dissolved in water, is loaded onto through pH
In the DE52 type resin chromatography post of the phosphate buffer pre-treatment of=7-9, static 12h, with distilled water, flow velocity is with 1-3 mL/
Min eluting, then through sephadex-G25 gel chromatography column, with distilled water, flow velocity, with 0.5-1 mL/min eluting, concentrates eluting
Liquid, then remove its polysaccharide pigment through activated carbon, it is vacuum dried to obtain white powder dicliptera chinensis polysaccharide.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108938720A (en) * | 2018-10-18 | 2018-12-07 | 郑老海 | A kind of Chinese medicine and preparation method thereof for treating diabetes |
| CN115677869A (en) * | 2022-09-14 | 2023-02-03 | 临沂大学 | Preparation method and application of golden ganoderma polysaccharide |
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| CN102283859A (en) * | 2011-09-07 | 2011-12-21 | 广东药学院 | Application of dicliptera chinensis polysaccharide |
| CN104083393A (en) * | 2014-04-01 | 2014-10-08 | 桂林医学院 | Application of dicliptera chinensis polysaccharide in preparation of medicines for treating liver injury caused by antituberculosis drugs |
| CN104189371A (en) * | 2014-09-02 | 2014-12-10 | 霍占祥 | Formula of traditional Chinese medicine for treating kidney yin depletion type diabetes |
| CN105638968A (en) * | 2014-10-22 | 2016-06-08 | 宾业崴 | Macrosolen cochinchinensis beverage tea |
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2016
- 2016-07-21 CN CN201610576684.9A patent/CN106265717B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102283859A (en) * | 2011-09-07 | 2011-12-21 | 广东药学院 | Application of dicliptera chinensis polysaccharide |
| CN104083393A (en) * | 2014-04-01 | 2014-10-08 | 桂林医学院 | Application of dicliptera chinensis polysaccharide in preparation of medicines for treating liver injury caused by antituberculosis drugs |
| CN104189371A (en) * | 2014-09-02 | 2014-12-10 | 霍占祥 | Formula of traditional Chinese medicine for treating kidney yin depletion type diabetes |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108938720A (en) * | 2018-10-18 | 2018-12-07 | 郑老海 | A kind of Chinese medicine and preparation method thereof for treating diabetes |
| CN115677869A (en) * | 2022-09-14 | 2023-02-03 | 临沂大学 | Preparation method and application of golden ganoderma polysaccharide |
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