CN106265717B - Application of the dicliptera chinensis polysaccharide in preparing prevention diabetes medicament or health products - Google Patents
Application of the dicliptera chinensis polysaccharide in preparing prevention diabetes medicament or health products Download PDFInfo
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- CN106265717B CN106265717B CN201610576684.9A CN201610576684A CN106265717B CN 106265717 B CN106265717 B CN 106265717B CN 201610576684 A CN201610576684 A CN 201610576684A CN 106265717 B CN106265717 B CN 106265717B
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- dicliptera chinensis
- polysaccharide
- dicliptera
- chinensis polysaccharide
- diabetes
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Classifications
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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
-
- 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)
Abstract
The present invention discloses application of the dicliptera chinensis polysaccharide in preparing prevention diabetes medicament or health products.By preparing diabetic animal experiment, it is found that it can improve the symptom of diabetes rat, inhibit dead, maintain weight, reduce blood glucose and improve resistance to sugar amount, show its good blood sugar reducing function.In addition, dicliptera chinensis polysaccharide can reduce diabetes rat serum glycosylation end products peptide (AGE P), glycosylated hemoglobin (HbAlc), total cholesterol（TC）, triglycerides（TG）, low-density lipoprotein（LDL‑C）With kidney aldose reductase（AR）Activity or content increase liver glycogen and high-density lipoprotein（HDL‑C）Content illustrates that it, by inhibiting liver, renal lesions and improving blood fat environment, has the function that inhibit diabetic complication.Being experimentally confirmed dicliptera chinensis polysaccharide has preferable prevention diabetes and its inhibits complication effect, and this new application is that its medical value has opened up frontier, has important application value and economic benefit.
Description
Technical field
The invention belongs to tcm field, be related to be with Dicliptera chinensis Dicliptera chinensis (L.) Nees. herbs
Raw material extraction purification goes out dicliptera chinensis polysaccharide and its application in being prepared into prevention diabetes medicament or health products,
Background technology
Dicliptera chinensis is the complete of Acanthaceae China peristrophe category Dicliptera chinensis Dicliptera chinensis (L.) Nees.
Grass.Herb is sweet in flavor, slight bitter, cold in nature, the thoughts of returning home, liver, lung channel, there is heat-clearing, cool blood, dampness removing, detoxify, promote the production of body fluid, diuresis and other effects, face
Bed be usually used in treating cold, fever, pyreticosis macular eruption, hot summer weather polydipsia, hematemesis blood, have blood in stool, hematuria, uterine bleeding, cough with lung heat, throat it is swollen
Bitterly, under the hot hot eyes of liver, eye conjunctivitis, child convulsion, dribbling urination, band, the cards such as herpes zoster, carbuncle swells malignant boil furuncle, snake dog bite.Dog
Liver dish resource is very abundant, the ground such as distribution Yunnan, Guangdong, Guangxi, for the civil medicinal herbs most in use in southwest.Dicliptera chinensis is Guangxi Folk medicine
Dual-purpose common strong medicine is eaten, the substances such as polysaccharide, organic acid, amino acid are mainly contained.Invention person's early-stage study finds dog
Liver dish polysaccharide component has anti-inflammatory, anti-oxidant and raising immunity of organism, reaches improvement and adjusts organism metabolism effect, but confrontation sugar
The research of urine disease still belongs to the first time.
Diabetes (Diabetes mellitus, DM) are often referred to diabetes B (TZDM), account for 90% or more of DM sums,
Its incidence, dead rate are high, worldwide have become the third-largest serious harm after cardiovascular and cerebrovascular disease and cancer
The disease of human health.Diabetes be by heredity and acquired environment collective effect, it is chronic based on carbohydrate metabolism disturbance
Comprehensive disease.With disease, can cause it is multigroup knit multiple organ lesion, seriously threaten human health.In traditional sugar
In urine disease treatment, diabetes treatment by western medicine is quick, but needs to take for a long time, and erious adverse reaction.Modern study is found, in part
Medicine or its extract have curative effect stable and adverse reaction is few, there is that Western medicine is irreplaceable in terms for the treatment of diabetes
And advantage.Therefore safe and effective antidiabetic medicine is found from natural drug to have a very important significance.Dicliptera chinensis medicine
With resourceful, the cost for obtaining dicliptera chinensis polysaccharide is low, suitable for mass production.The research of domestic and international dicliptera chinensis polysaccharide is retrieved, still
Have no the research report prevented dicliptera chinensis polysaccharide in terms of diabetes effect.
Invention content
The purpose of the present invention is going out dicliptera chinensis polysaccharide by raw material extraction purification of Dicliptera chinensis herb, and it is prepared into preventing
Diabetes medicament or health products, this announcement that still belongs to the first time at home.
Realizing the technical solution of the object of the invention is：
The new application of dicliptera chinensis polysaccharide is used to prepare the drug or health products of prevention diabetes, can be prepared into for mouth
The Chinese medicine preparation of clothes, such as powder, tablet, oral solution, granule, capsule and pill etc., 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 Dicliptera chinensis herb coarse powder, and petroleum ether reflux degreasing is added and carries
1-2h, the dregs of a decoction is taken to volatilize after petroleum ether to the greatest extent, extracting in boiling water 1-3 times, each 1-3h merges simultaneously concentrated extracting solution, with chloroform and just
The ratio 4 of butanol:1, using Sevag fado time extractions, the supernatant for the protein that is removed adds absolute ethyl alcohol that alcohol content is made to reach
To 70-99%, the alcohol precipitation in 4 DEG C of refrigerators filters, discards supernatant liquid, filter residue is washed with ether, absolute ethyl alcohol, acetone successively, i.e.,
Obtain Dicliptera chinensis Thick many candies；Dicliptera chinensis Thick many candies are dissolved in water, are loaded onto the DE52 of the phosphate buffer pre-treatment through pH=7-9
In type resin chromatography column, static 12h, with distilled water, flow velocity is eluted with 1-3mL/min, then through sephadex-G25 gel chromatographies
Column, with distilled water, flow velocity is eluted with 0.5-1mL/min, concentrate eluant, then removes its polysaccharide pigment through activated carbon, and vacuum is dry
It is dry to obtain white powder dicliptera chinensis polysaccharide (DCP).
Compared with prior art, the present invention Dicliptera chinensis herb resource is abundant, prepare that dicliptera chinensis polysaccharide is simple for process, cost
It is low, it is without any side effects, it is in good taste, there is good Development volue.It is suitable in addition, the present invention has excavated dicliptera chinensis polysaccharide
Diabetes are prevented, it is curative for effect, there is good medical value preparing prevention diabetes medicament or health products, and be Dicliptera chinensis
Polysaccharide further deeply develops offer Research foundation.
Description of the drawings
Fig. 1 is influence (n=10) of the dicliptera chinensis polysaccharide of the embodiment of the present invention to diabetes rat body weight；
Fig. 2 is influence of the dicliptera chinensis polysaccharide of the embodiment of the present invention to blood glucose in diabetic rats concentration；
Fig. 3 is influence of the dicliptera chinensis polysaccharide of the embodiment of the present invention to diabetes rat sugar tolerance；
In figure：A, correspondence represents blank control group, model group, the positive (glibenclamide) group, Dicliptera chinensis respectively by B, C, D, E, F
The high, medium and low dosage group of polysaccharide,
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).
Specific implementation mode
The content of present invention is further described with reference to embodiment and attached drawing, but is not limitation of the invention.
One, it is the extraction and purification method of dicliptera chinensis polysaccharide below：
Dry Dicliptera chinensis herb coarse powder 2000g is weighed, the reflux degreasing of 20000ML petroleum ethers is added and extracts 2h.The dregs of a decoction are waved
To the greatest extent after petroleum ether, extracting in boiling water 2 times, each 1.5h merges extracting solution, 200mL is concentrated into, with chloroform-n-butanol (4:1) repeatedly
It extracts (i.e. Sevag methods), the supernatant for the protein that is removed adds 95% ethyl alcohol that alcohol content is made to reach 80%, in 4 DEG C of refrigerators
Alcohol precipitation 12h.It filters, discards supernatant liquid, filter residue is washed with ether, absolute ethyl alcohol, acetone to get Dicliptera chinensis Thick many candies successively.It will
Dicliptera chinensis Thick many candies are dissolved in water, are loaded onto in the DE52 type resin chromatography columns of the phosphate buffer pre-treatment through PH=8,
Static 12h, with distilled water, flow velocity is eluted with 2mL/min, then through sephadex-G25 gel chromatography columns, with distilled water, flow velocity with
0.5-1mL/min is eluted, concentrate eluant, then removes its polysaccharide pigment through activated carbon, is dried in vacuo to obtain white powder dog liver
Dish polysaccharide (DCP).Dicliptera chinensis polysaccharide compares product with glucose, using one sulfuric acid method of phenol, through UV-vis spectroscopy meter
Purity of polysaccharide in determination sample is up to 95.6%.
Two, dicliptera chinensis polysaccharide is prepared into pharmaceutical preparation：
Corresponding auxiliary material can be added in dicliptera chinensis polysaccharide, is prepared into powder, tablet, oral solution, granule, capsule and pill
Deng.
Three, dicliptera chinensis polysaccharide to diabetes rat hypoglycemic and inhibits complication effect experiment method and result：
1, material, reagent and instrument
1.1 animal
SPF grades of SD rats 90, half male and half female provide (credit number by Medical Colleges Of Guilin's SPF Experimental Animal Centers
SCXK2007-0001)。
1.2 drugs and reagent
Dicliptera chinensis is accredited as Acanthaceae China peristrophe category Dicliptera chinensis through Guangxi University of Chinese Medicine professor Wei Songji
The herb of Dicliptera chinensis (L.) Nees..Reality is prepared with " the extraction and purification method of dicliptera chinensis polysaccharide "
It tests with dicliptera chinensis polysaccharide (DCP) for reagent product.
Glibenclamide, Huarun Zizhu Pharmaceutical Co., Ltd..Streptozotocin (STZ), is purchased from Sigma companies.Total cholesterol
(TC), triglycerides (TG), high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C), are purchased from Nanjing and build up biological work
Journey research institute.Insulin radioimmunoassay immune reagent kit is purchased from Bio-Rad companies.One peptide of glycosylation end products (AGE-P), liver glycogen,
Kidney aldose reductase (AR) and glycosylated hemoglobin (HbAlc) detection kit, are purchased from the limited public affairs of Zhejiang Ai Kang biotechnologies
Department.
1.3 instrument
TLL-C tabletop refrigerated centrifuges, Beijing Fourth Ring scientific instrument factory；TSN-695B types, which are put, exempts from measuring instrument, Shanghai day ring
One factory of instrument；AU600 full-automatic biochemical detectors, Olympus production；752 type spectrophotometers, Shanghai third analytical instrument
Factory；BS1105 electronic balances, German sartorius companies；Bayer blood glucose monitoring system and its test paper, German Bayer.
2. experimental method
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 are fed with high glucose and high fat feed 4 weeks.All Rat Fasts can't help water 12h after 4 weeks, and modeling group is noted by the abdominal cavities 35mg/kg
Streptozotocin (STZ) modeling is penetrated, Normal group only injects equivalent lemon acid-sodium citrate buffer solution (pH=4.4), after injection
Continue to be deprived of food but not water 12h.After feeding 72h again, it is deprived of food but not water 12h, the fasting blood-glucose of modeling group, sieve are measured with blood glucose meter
It is the successful diabetes rat of modeling to select fasting blood-glucose >=13.8mmol/L, this experiment is at mould 74.By the successful rat of modeling
It is divided into model group, positive controls (glibenclamide, 20mg/kg) and dicliptera chinensis polysaccharide according to blood glucose value and weight stochastic averagina
High, medium and low dosage group (600,400,200mg/kg), gastric infusion, Normal group and model group gavage distilled water, it is continuous to fill
Stomach totally 6 weeks.The general status that animal is observed during experiment is weighed 1 time in each week before administration and after administration, and quiet through animal tail
Arteries and veins is taken a blood sample, and whole blood blood glucose is measured.After being administered 6 weeks, it is deprived of food but not water 12h, gavage gives glucose 2.5g/ to each group rat respectively
Kg draws change of blood sugar figure respectively at 0h, 0.5h, 1h, 2h, 3h tail vein measuring blood sugar of blood extracting value, and t examines more a time
The glucose-tolerant degree of point.After being deprived of food but not water 4h again, chloral hydrate anesthesia, abdominal aortic blood takes liver and kidney.
3. experimental result
The observation of 3.1 rat general status
The normal rats state of mind is good, and weight gain moves freely.Diabetic model group rat occurs typical more
Food, more drink, diuresis, weight loss symptoms, and tired mind, activity are reduced, hair is lackluster, 4 death during experiment；Dog
Liver dish polysaccharide group rat amount of drinking water, urine volume, dietary amount are substantially reduced compared with model group, and weight is dramatically increased compared with diabetes rat,
Only there is 1 death in low dosage, illustrates that dicliptera chinensis polysaccharide can improve the symptom of diabetes rat.
The variation of 3.2 rat body weights
After rats by intraperitoneal injection STZ 72h, each modeling group and modeling precursor heavy phase ratio are remarkably decreased (P ＜ 0.01).It is real
During testing, blank group weight gradually increases, and modeling group rat is in reduction trend, under wherein model group rats weight is substantially in
Drop trend, declines in the last fortnight especially after modeling particularly evident, and medication group pharmaceutical intervention is after two weeks, positive group and dog liver
Weight is substantially in rising trend from the 2nd week for dish polysaccharide high dose group, in dicliptera chinensis polysaccharide, low dose group weight keep stablize.
See Fig. 1.
The variation of 3.3 rat blood sugars level
Before administration, compared with blank group blood glucose level, modeling group is statistically significant (P ＜ 0.01), illustrate modeling at
Work(.After administration, from the 2nd week, compared with model group, each dosage group of dicliptera chinensis polysaccharide and positive group blood glucose are substantially reduced, and are had
Statistical significance (P ＜ 0.05).See Fig. 2.
3.4 oral glucose tolerances (OGTT) are tested
OGTT experiments are clinically to diagnose the goldstandard of DM.Experimental result is shown, compared with model group, positive group and dog liver
The high, medium and low dosage group of dish polysaccharide can significantly improve diabetes rat glucose tolerance, the blood when gavage gives glucose 3h
Sugared value difference is heteropolar significantly.See Fig. 3.
3.5 dicliptera chinensis polysaccharides are to diabetes rat serum advanced glycosylation end products-peptide (AGE-P) and glycosylated hemoglobin
(HbAlc) influence
Monitor the screening that advanced glycosylation end products-peptide (AGE-P) is used for DM people at highest risk, glycosylated hemoglobin (HbAlc) row
For one of the main standard of DM and the diagnosis of its complication.Experimental result is shown, compared with model group, medication group rat AGE-P and
The significant reductions of HbAlc, show that dicliptera chinensis polysaccharide can improve diabetes and its complication, the result table 1 and Fig. 2 results
It is consistent.
Influence of 1 dicliptera chinensis polysaccharide of table (DCP) to diabetes rat AGE-P and HbAlc
Compared with model group, * P ＜ 0.05, * * P ＜ 0.01 (following table indicates and dosage is same).
3.6 dicliptera chinensis polysaccharides are to diabetes rat hepatic tissue liver glycogen, kidney aldose reductase (AR) activity and level
It influences
The effect of liver glycogen is to maintain stablizing relatively for blood sugar concentration, and under hyperglycemic state (such as diabetes), hexose swashs
Enzyme is saturated, at this moment aldose reduction enzyme activition, promotes internal renal lesions.Experimental result is shown, compared with model group, medication
Group rat liver glycogen increases and kidney AR conspicuousnesses reduce, and shows that dicliptera chinensis polysaccharide can regulate and control diabetes glucose concentration and inhibition
The complication of kidney.It is shown in Table 2.
2 dicliptera chinensis polysaccharide of table (DCP) is to the influence to diabetes rat liver glycogen and A R
Influence of 3.7 dicliptera chinensis polysaccharides to blood fat factor TC, TG, HDL-C and LDL-C levels in diabetes rat serum
Compared with blank group, the horizontal conspicuousness of model group rats TC, TG, LDL-C increases, the horizontal conspicuousness drops of HDL-C
It is low, show that serious diseases occurs in the blood lipid metabolism of diabetes rat.By 6 weeks pharmaceutical interventions, compared with model group, Dicliptera chinensis
The horizontal conspicuousnesses of each dosage group rat TC, TG, LDL-C of polysaccharide reduce, and the horizontal conspicuousnesses of HDL-C increase, and show that Dicliptera chinensis is more
Sugar can have improvement result to the blood fat environment of diabetes rat.It is shown in Table 3.
Influence of 3 dicliptera chinensis polysaccharide of table (DCP) to LDL-C, HDL-C, TG and TC in diabetes rat serum
By testing above, it is found that dicliptera chinensis polysaccharide can improve the symptom of diabetes rat, inhibit dead, maintain body
Weight reduces blood glucose and improves resistance to sugar amount, shows its good blood sugar reducing function.In addition, can to reduce diabetes big for dicliptera chinensis polysaccharide
It is mouse serum advanced glycosylation end products-peptide (AGE-P), glycosylated hemoglobin (HbAlc), total cholesterol (TC), triglycerides (TG), low
Density lipoprotein (LDL-C) and kidney aldose reductase (AR) activity or content, increase liver glycogen and high-density lipoprotein (HDL-
C) content illustrates that it, by inhibiting liver, renal lesions and improving blood fat environment, has the function that inhibit diabetic complication.
Claims (2)
1. application of the dicliptera chinensis polysaccharide in preparing prevention diabetes medicament or health products.
2. application according to claim 1, it is characterized in that：The preparation of the dicliptera chinensis polysaccharide is to weigh dry Dicliptera chinensis
Herb coarse powder is added petroleum ether reflux degreasing and extracts 1-2h, and the dregs of a decoction volatilize after petroleum ether to the greatest extent, extracting in boiling water 1-3 times, each 1-
3h merges simultaneously concentrated extracting solution, with the ratio 4 of chloroform and n-butanol:1, using Sevag fado time extractions, be removed albumen
The supernatant of matter, adds absolute ethyl alcohol that alcohol content is made to reach 70-99%, the alcohol precipitation in 4 DEG C of refrigerators, filters, discards supernatant liquid, filter residue
It is washed successively to get Dicliptera chinensis Thick many candies with ether, absolute ethyl alcohol, acetone；Dicliptera chinensis Thick many candies are dissolved in water, are loaded onto through pH
In the DE52 type resin chromatography columns of the phosphate buffer pre-treatment of=7-9, static 12h, with distilled water, flow velocity is with 1-3 mL/
Min is eluted, then through sephadex-G25 gel chromatography columns, and with distilled water, flow velocity is eluted with 0.5-1 mL/min, concentration elution
Liquid, then its polysaccharide pigment is removed through activated carbon, it is dried in vacuo to obtain white powder dicliptera chinensis polysaccharide.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
Citations (4)
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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 |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (1)
| Title |
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| 狗肝菜多糖保肝作用研究;李海燕等;《中药材》;20060831;第29卷(第08期);833-834 * |
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