CN108685938A - The hypoglycemic purposes of beans taro leaf polyose - Google Patents
The hypoglycemic purposes of beans taro leaf polyose Download PDFInfo
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- CN108685938A CN108685938A CN201810404020.3A CN201810404020A CN108685938A CN 108685938 A CN108685938 A CN 108685938A CN 201810404020 A CN201810404020 A CN 201810404020A CN 108685938 A CN108685938 A CN 108685938A
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- beans taro
- taro leaf
- leaf polyose
- beans
- polyose
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- 244000205754 Colocasia esculenta Species 0.000 title claims abstract description 94
- 235000006481 Colocasia esculenta Nutrition 0.000 title claims abstract description 94
- 244000046052 Phaseolus vulgaris Species 0.000 title claims abstract description 92
- 235000010627 Phaseolus vulgaris Nutrition 0.000 title claims abstract description 92
- 230000002218 hypoglycaemic effect Effects 0.000 title claims abstract description 19
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims abstract description 36
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 31
- 239000008103 glucose Substances 0.000 claims abstract description 30
- 102000004877 Insulin Human genes 0.000 claims abstract description 18
- 108090001061 Insulin Proteins 0.000 claims abstract description 18
- 229940125396 insulin Drugs 0.000 claims abstract description 18
- 239000003814 drug Substances 0.000 claims abstract description 15
- 229940079593 drug Drugs 0.000 claims abstract description 10
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 7
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 7
- 230000004190 glucose uptake Effects 0.000 claims description 4
- 230000000694 effects Effects 0.000 abstract description 15
- 235000013305 food Nutrition 0.000 abstract description 7
- 206010012601 diabetes mellitus Diseases 0.000 abstract description 5
- 208000017667 Chronic Disease Diseases 0.000 abstract description 3
- 238000011156 evaluation Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 16
- 239000001963 growth medium Substances 0.000 description 13
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 150000004676 glycans Chemical class 0.000 description 10
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 235000009508 confectionery Nutrition 0.000 description 7
- 239000013641 positive control Substances 0.000 description 7
- 230000004663 cell proliferation Effects 0.000 description 6
- 230000004700 cellular uptake Effects 0.000 description 6
- 229920001282 polysaccharide Polymers 0.000 description 6
- 239000005017 polysaccharide Substances 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 239000012980 RPMI-1640 medium Substances 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 235000019441 ethanol Nutrition 0.000 description 5
- 238000011081 inoculation Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 4
- 239000012141 concentrate Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 150000004804 polysaccharides Polymers 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 239000012045 crude solution Substances 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 230000001771 impaired effect Effects 0.000 description 3
- 230000031700 light absorption Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000012549 training Methods 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 2
- 244000100170 Phaseolus lunatus Species 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003636 conditioned culture medium Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000469 ethanolic extract Substances 0.000 description 2
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 2
- 150000002515 isoflavone derivatives Chemical class 0.000 description 2
- 235000008696 isoflavones Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 2
- 229960003105 metformin Drugs 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
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- 210000002966 serum Anatomy 0.000 description 2
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- 239000011782 vitamin Substances 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- QUTFFEUUGHUPQC-ILWYWAAHSA-N (2r,3r,4s,5r)-3,4,5,6-tetrahydroxy-2-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]hexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](C=O)NC1=CC=C([N+]([O-])=O)C2=NON=C12 QUTFFEUUGHUPQC-ILWYWAAHSA-N 0.000 description 1
- 241000913828 Apios Species 0.000 description 1
- 244000176051 Apios tuberosa Species 0.000 description 1
- 235000003276 Apios tuberosa Nutrition 0.000 description 1
- 241001269238 Data Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 230000023852 carbohydrate metabolic process Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 210000004153 islets of langerhan Anatomy 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000005360 mashing Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000022558 protein metabolic process Effects 0.000 description 1
- 230000003946 protein process Effects 0.000 description 1
- 235000021251 pulses Nutrition 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
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- 238000002525 ultrasonication Methods 0.000 description 1
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Classifications
-
- 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
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Food Science & Technology (AREA)
- Obesity (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hematology (AREA)
- Nutrition Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of hypoglycemic purposes of beans taro leaf polyose, and developing natural antihypelipidemic product for fields such as medicine, Food Sciences provides new resources.HepG2 cell models, RINm5F cell model experimental evaluations through high sugar induced structure in the present invention prove, beans taro leaf polyose can significantly improve the ability of HepG2 cell consumptions intake glucose, promote the synthesis of RINm5F cells and excreting insulin, function and effect comprehensively and notable.Therefore, beans taro leaf polyose of the invention can be applied to prepare antihypelipidemic foodstuff or drug, be occurred with treating and preventing diabetes and its chronic diseases.
Description
Technical field
The present invention relates to field of medicaments, and in particular to a kind of hypoglycemic purposes of beans taro leaf polyose.
Background technology
U.S. beans taro (Apios americana Medikus) is mainly grown in the Ontario, Canada in North America to the U.S.
Between Florida, there is extensive cultivated area in American-European and Japan at present.This is a kind of perennial pulse family Root of Fortune Apios category plant
Object is similar to potato.Edible part is underground stem tuber, is a kind of high protein, high-calcium food, thus by American Indian as leading
Food.U.S. beans taro introduced China's plantation in 2009, and was cultivated well.Research shows that its flower, leaf, stem tuber are containing rich
The ingredients such as rich free amino acid, soluble protein, flavones, isoflavones, saponin(e, vitamin and mineral.Wherein, beans taro contains
There are the vitamin, total saposins and isoflavones of high level, there is great potentiality to be exploited.
According to epidemiological analysis, diabetes are a kind of chronic diseases having long history.It can severely impact the mankind
Eubolism, such as carbohydrate, lipid and protein metabolism.This metabolic disorder be typically by insulin secretion not
Caused by foot or insulin inactivation.Common symptom has:Diuresis, more drinks, more food, weight loss etc., can threat to life when serious.
2000 annual datas statistics shows the torment that the whole world has 1.91 hundred million populations enduring diabetes, it is contemplated that the year two thousand thirty, this number will
Reach 3.06 hundred million.Although this is that a kind of coverage is wide, the quite deep disease of harm, since it is practised by heredity, diet, life
The used influence for waiting many factors, we are difficult to start in terms of single, find the not high precautionary measures of effective, convenient and cost
Therapeutic scheme.How prevention and treatment diabetes and its related complication have caused the concern of whole world scholar, natural plants to be lived
Property substance is particularly subject to favor, but the function of polysaccharide of beans taro leaf polyose is rarely reported at present.
Invention content
The technical problem to be solved in the present invention is to provide a kind of hypoglycemic purposes of beans taro leaf polyose, are medicine, Food Science
Equal fields develop natural antihypelipidemic product and provide new resources.
In order to solve the above technical problems, the present invention provides a kind of hypoglycemic purposes of beans taro leaf polyose:
It is used to prepare antihypelipidemic foodstuff and/or drug.
The improvement of hypoglycemic purposes as beans taro leaf polyose of the present invention:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced HepG2 cells to glucose consumption ability.
Hypoglycemic purposes as beans taro leaf polyose of the present invention is further improved:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced HepG2 cells to glucose uptake ability.
Hypoglycemic purposes as beans taro leaf polyose of the present invention is further improved:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced RINm5F cells synthesis excreting insulin ability.
Note:Above-mentioned beans taro leaf polyose is obtained by prior art preparation, such as can be prepared by following steps:
1), the preparation of beans taro leaf polyose crude product:
According to 1g:The solid-liquid ratio of 10ml weighs beans taro leaf 100g and 1000mL distilled water is added, and 80 DEG C of water-bath 4h are extracted
(enzyme deactivation is also achieved while extraction), is then beaten, then centrifuges (4000r/min centrifuges 30min), by the supernatant of centrifugation gained
Liquid filters and collects filtrate;
It is substituted after beans taro leaf repeats above-mentioned addition distilled water with the filter residue of centrifugation gained and first carries out extraction process (80 DEG C of water-baths
4h) and then mashing, the process that centrifuges again 2 times, merge 3 centrifugations and filter the filtrate concentrated by rotary evaporations of gained to the 1/5 of original volume,
Obtain concentrate.
Under stirring condition, ethyl alcohol (straight alcohol) is added in the concentrate of gained to ethyl alcohol final concentration of 85%, at 4 DEG C
It stands and collects precipitation afterwards for 24 hours, obtain beans taro leaf Thick many candies;
2), the purifying of beans taro leaf Thick many candies, follows the steps below successively:
A, activated carbon decolorizing:
By beans taro leaf Thick many candies be placed in beaker with distillation water dissolution (dosage of distilled water need to only can guarantee that beans taro leaf is slightly more
Sugar dissolving), beans taro leaf Thick many candies solution is obtained, activated carbon is added in beans taro leaf Thick many candies solution in 80 ± 10 DEG C of water-baths
It is filtered after middle stirring 45min;The mass ratio of beans taro leaf Thick many candies and activated carbon is 15:1;
It is substituted after above-mentioned beans taro leaf Thick many candies solution repeats above-mentioned addition activated carbon and stirring and is filtered with the filtrate of gained
Decolorization 3 times, the filtrate of final gained is in colourless, polysaccharide crude solution after must decolourizing at this time;
B, Sevage methods take off albumen:
According to 4:100mL Sevage reagents, Yu Ci is added in polysaccharide crude solution after 400mL decolorations in 1 volume ratio
1000 turns/min rotating speeds stir 30min in power blender, are then centrifuged for, and water layer and position positioned at lower layer are removed by separatory funnel
Albuminate between the solvent layer on upper layer;The Sevage reagents are by n-butanol:Chloroform=1:5 volume ratio mixing and
;
Polysaccharide crude solution repeats above-mentioned addition after substituting above-mentioned decoloration with the gains after the removing albuminate of gained
Sevage reagents, magnetic agitation, centrifugation remove the de- protein process 5 times of albuminate, must remove the polysaccharide solution of protein;
The polysaccharide solution concentrated by rotary evaporation for removing protein is obtained into concentrate (about the 10% of original volume), by concentrate
In -80 DEG C of pre-freeze 6h, then use vacuum freeze drier 48h (technological parameter that vacuum freeze drier is set as -40 DEG C,
It 1.2Pa) is dried into powdered (moisture content≤0.1%), obtains beans taro leaf polyose.
The invention discloses a kind of hypoglycemic purposes of beans taro leaf polyose, are the natural drop of the fields such as medicine, Food Science exploitation
Sugar product provides new resources.HepG2 cell models, RINm5F cell model experimental evaluations through high sugar induced structure in the present invention
It proves, beans taro leaf polyose can significantly improve the ability of HepG2 cellular uptake consumption of glucose, promote the synthesis of RINm5F cells simultaneously
Excreting insulin, function and effect are comprehensively and notable.Therefore, beans taro leaf polyose of the invention be applied to prepare antihypelipidemic foodstuff and/or
Drug is occurred with treating and preventing diabetes and its chronic diseases.
The usage of the beans taro leaf polyose of the present invention is oral, dosage about 150~250mg every time, three times a day.
The present invention compared with the existing technology, has the following advantages:
1. present invention firstly discovers that beans taro leaf polyose can promote high sugar induced HepG2 cellular uptake consumption of glucose, table
Reveal significant hypoglycemic effect, there is good development prospect.
2. present invention firstly discovers that beans taro leaf polyose can promote high sugar induced RINm5F cells to synthesize excreting insulin, tool
There is good development prospect.
3. the present invention provides new medical application for beans taro leaf polyose, a new application field has been expanded.
Description of the drawings
Fig. 1 is influence of the various concentration beans taro leaf polyose to HepG2 cell Proliferations in the present invention.
Fig. 2 is influence of the various concentration beans taro leaf polyose to HepG2 cell consumption glucose abilities in the present invention.
Fig. 3 is that (fluorescence shines for influence of the various concentration beans taro leaf polyose to HepG2 cellular uptake glucose abilities in the present invention
Piece).
Fig. 4 is that (fluorescence is fixed for influence of the various concentration beans taro leaf polyose to HepG2 cellular uptake glucose abilities in the present invention
Amount).
Fig. 5 is influence of the various concentration beans taro leaf polyose to RINm5F cell Proliferations in the present invention.
Fig. 6 is influence of the various concentration beans taro leaf polyose to RINm5F insulin inside cells contents in the present invention.
Fig. 7 is influence of the various concentration beans taro leaf polyose to the extracellular insulin contents of RINm5F in the present invention.
Note:In Fig. 2~Fig. 4 of the present invention:
C represents low sugar control group (DMEM concentration of glucose 5.5mM);
H represents high sugared control group (DMEM concentration of glucose 30mM);
M represents positive controls (DMEM concentration of glucose 30mM+2mM melbine)
In Fig. 6~Fig. 7 of the present invention:
C represents low sugar control group (RPMI-1640 concentration of glucose 4.5mM);
H represents high sugared control group (RPMI-1640 concentration of glucose 16.7mM).
Specific implementation mode
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in
This:
The hypoglycemic purposes of embodiment 1, beans taro leaf polyose is intervened with beans taro leaf polyose by high sugar as Figure 1-Figure 4
The glucose uptake of the HepG2 cells of induction, observation HepG2 cells consumes ability, illustrates that beans taro leaf polyose has significant drop
Sugared effect.
Experiment 1, influence of the beans taro leaf polyose to HepG2 cell Proliferation vigor;
The HepG2 cell inoculations for collecting logarithmic phase are placed in 37 DEG C, 5%CO in 96 orifice plates per 5000, hole cell2Culture
It is incubated in case and is grouped afterwards for 24 hours.
It is divided into following 9 groups:Blank control group and 5,10,20,40,80,160,320 and 640ug/mL beans taro leaf polyoses
Group;Above-mentioned 5,10,20,40,80,160,320 and 640ug/mL beans taro leaf polyose groups are separately added into the ethyl alcohol extraction of corresponding concentration
Object is added without ethanol extract in blank control group.
Every group of 6 hole of repetition, is placed in 37 DEG C, 5%CO2After being incubated for 24 hours jointly in incubator, residual liquid in each hole is sucked out,
The MTT (0.5mg/mL is dissolved in not serum-containing media) of 100ul is added afterwards twice with the rinsing of phosphate buffer (PBS) solution,
In 37 DEG C, 5%CO2Continue culture 4 hours in incubator.
Then, it removes per boreliquid, 250ul dimethyl sulfoxide (DMSO)s (DMSO) is added per hole, 10 points are shaken on horizontal shaker
Clock, microplate reader measure its light absorption value at 570nm.Cell-proliferation activity (%)=experimental port OD values/control wells OD values * 100%;
According to Fig. 1, MTT experiment is not the results show that light absorption value has significant difference between each group, illustrate beans taro leaf polyose 0~
It is non-toxic to HepG2 cells in 640ug/mL concentration ranges, the proliferation of HepG2 cells had both been pressed down without facilitation effect or not
Effect processed.
Experiment 2, influence of the beans taro leaf polyose to HepG2 cell consumption glucose abilities;
The HepG2 cell inoculations of logarithmic phase are collected in 6cm culture dishes (density 4*105A cell/ware), low sugar training is added
Base (DMEM concentration of glucose 5.5mM) is supported, 37 DEG C, 5%CO are placed in2It is grouped after being incubated overnight in incubator;It is equally divided into following 5
Group:Low sugar control group, high sugared control group, melbine positive controls (a concentration of 2mM) and 100ug/mL beans taro leaf polyoses
Group and 300ug/mL beans taro leaf polyose groups, each group repeat 5 wares.
Wherein melbine positive controls are added melbine and are pre-processed for 24 hours in incubator.100ug/ml beans taro leaves
Polysaccharide group and 300ug/ml beans taro leaf polyose groups are separately added into corresponding concentration beans taro leaf polyose and are pre-processed for 24 hours in incubator.
Replace culture medium:It discards original culture solution and is cleaned twice with PBS, wherein low sugar control group adds low sugar culture medium
(DMEM concentration of glucose 5.5mM), remaining group increase sugar culture-medium (DMEM concentration of glucose 30mM).Keep diformazan double simultaneously
Determination of metformin and consistent, 100ug/mL beans taro leaf polyose group and 300ug/mL beans taro leaves when pretreatment in guanidine positive controls
In polysaccharide group, beans taro leaf polyose concentration is consistent when being pre-processed with it.
By above-mentioned each group in incubator coprocessing for 24 hours after, replace culture medium:Discard original culture solution in culture dish
It is used in combination PBS to clean twice, changes low sugar culture medium into later.12h is cultivated in incubator.
Cell conditioned medium is collected, the glucose utilization in culture solution is measured using glucose kit.Discard extra training
Nutrient solution is simultaneously cleaned twice with PBS, and 1ml cell pyrolysis liquids are added in each culture dish, 2ml centrifuge tubes are collected in cell scraper.Ultrasound
Afterwards quantitative and corrected glucose consumption data are carried out with BCA protein reagents box.
According to Fig.2, glucose consumption experimental result is shown, the beans taro leaf polyose of 100ug/ml and 300ug/ml concentration
It can obviously repair by the glucose absorption ability of the lower impaired HepG2 cells of high sugar effect, and 100ug/ml and 300ug/ml
The beans taro leaf polyose effect of concentration is similar.
Experiment 3, influence of the beans taro leaf polyose to HepG2 cellular uptake glucose abilities;
The HepG2 cell inoculations of collection logarithmic phase (density 4*10 in 24 orifice plates5A cells/well), low sugar training is added
Support base (DMEM concentration of glucose 5.5mM), be placed in 37 DEG C, be incubated overnight in 5%CO2 incubators after be grouped;It is respectively divided into following
Five groups:Low sugar control group, high sugared control group, melbine positive controls (a concentration of 2mM), 100ug/mL beans taro leaf polyose groups
With 300ug/mL beans taro leaf polyose groups, each group repeats 3 holes.
Wherein melbine positive controls are added melbine and are pre-processed for 24 hours in incubator.100ug/ml beans taro leaves
Polysaccharide group and 300ug/ml beans taro leaf polyose groups are separately added into corresponding concentration beans taro leaf polyose and are pre-processed for 24 hours in incubator.
Replace culture medium:It discards original culture solution and is cleaned twice with PBS, wherein low sugar control group adds low sugar culture medium
(DMEM concentration of glucose 5.5mM), remaining group increase sugar culture-medium (DMEM concentration of glucose 30mM).Keep diformazan double simultaneously
Determination of metformin and consistent, 100ug/mL beans taro leaf polyose group and 300ug/mL beans taro leaves when pretreatment in guanidine positive controls
It is consistent when beans taro leaf polyose concentration is pre-processed with it in polysaccharide group.
In incubator coprocessing for 24 hours after, discard culture medium and cleaned twice with PBS, change into 2-NBDG containing 0.1mM spy
The fresh culture of needle and 100nM insulin, cultivates 30min in incubator.After dyestuff is clean simultaneously with fluorescence microscope
It takes pictures, calculates average optical density value.
According to fig. 3 and shown in Fig. 4, glucose uptake experimental result is shown, compared to low sugar condition, under high sugar effect,
HepG2 cells can absorb more glucose, but gap unobvious.Beans taro leaf polyose can improve HepG2 cellular uptakes Portugal
The ability of grape sugar.Wherein, compared to high sugared control group, the function and effect of the beans taro leaf polyose (300ug/ml) of high concentration are very bright
It is aobvious.
Embodiment 2, as shown in Figure 5-Figure 7, intervened the RINm5F cells by high sugar induced with beans taro leaf polyose, observe
The insulin synthesis of RINm5F cells secretes situation, illustrates that beans taro leaf polyose has significant hypoglycemic effect.
Experiment 1, influence of the beans taro leaf polyose to RINm5F cell Proliferation vigor;
The RINm5F cell inoculations of logarithmic phase are collected in 96 orifice plates, per 5000, hole cell, are placed in 37 DEG C, 5%CO2 cultures
It is incubated in case and is grouped afterwards for 24 hours.
It is divided into following 9 groups:Blank control group and 5,10,20,40,80,160,320 and 640ug/mL beans taro leaf polyoses
Group;Above-mentioned 5,10,20,40,80,160,320 and 640ug/mL beans taro leaf polyose groups are separately added into the ethyl alcohol extraction of corresponding concentration
Object is added without ethanol extract in blank control group.
Every group of 6 hole of repetition, is placed in 37 DEG C, 5%CO2After being incubated for 24 hours jointly in incubator, residual liquid in each hole is sucked out,
The MTT (0.5mg/mL is dissolved in not serum-containing media) of 100ul is added afterwards twice with the rinsing of phosphate buffer (PBS) solution,
In 37 DEG C, 5%CO2Continue culture 4 hours in incubator.
Then, it removes per boreliquid, 250ul dimethyl sulfoxide (DMSO)s (DMSO) is added per hole, 10 points are shaken on horizontal shaker
Clock, microplate reader measure its light absorption value at 570nm.Cell-proliferation activity (%)=experimental port OD values/control wells OD values * 100%;
According to Fig.5, MTT experiment is the results show that compared with blank control group, and the beans taro leaf polyose of 40~640ug/ml concentration is to thin
Born of the same parents' proliferation activity has apparent facilitation.Beans taro leaf polyose is nontoxic to RINm5F cells in 0-640ug/mL concentration ranges
Property.
Experiment 2, beans taro leaf polyose synthesize RINm5F cells the influence of excreting insulin;
The RINm5F cell inoculations of logarithmic phase are collected in 6cm culture dishes (density 4*105A cell/ware), low sugar is added
Culture medium (RPMI-1640 concentration of glucose 4.5mM), be placed in 37 DEG C, be incubated overnight in 5%CO2 incubators after be grouped, that is, point
For low sugar control group, high sugared control group, 200ug/mL beans taro leaf polyose group and 300ug/mL beans taro leaf polyose groups.Wherein 200ug/
The beans taro leaf polyose that corresponding concentration is added in mL beans taro leaf polyose group and 300ug/ml beans taro leaf polyose groups pre-processes in incubator
24h。
Replace culture medium:It discards original culture solution in hole and is cleaned twice with PBS, wherein low sugar control group adds low sugar to train
Base (RPMI-1640 concentration of glucose 4.5mM) is supported, remaining increases sugar culture-medium (RPMI-1640 concentration of glucose 16.7mM) simultaneously
The beans taro leaf polyose concentration for keeping beans taro leaf polyose group consistent with when pretreatment.In incubator coprocessing for 24 hours after, replace culture
Base:It discards original culture solution in culture dish and is cleaned twice with PBS, change low sugar culture medium into without exception.It is cultivated in incubator
12h。
Cell conditioned medium is collected, the insulin content in Elisa kit measurement culture solutions is used.Discard extra culture solution
It is used in combination PBS to clean twice, 1mlPBS is added in each ware, 2ml centrifuge tubes are collected in cell scraper.A part of use is taken after ultrasonication
BCA protein reagent boxes are quantified, and Elisa kit measurement insulin contents are used after rest part centrifugation.By intraor extracellular
Insulin content is corrected with albumen concentration.
According to Fig. 6,7, the results show that under high sugar effect, the synthesis of RINm5F cells divides insulin content determination experiment
The ability for secreting insulin is impaired.200,300 and 400ug/ml beans taros leaf polyose can obviously be repaired lower impaired by high sugar effect
RINm5F cells synthesis and excreting insulin ability, significant effect.Show that beans taro leaf polyose can promote islet cells to increase
Excreting insulin is grown and synthesizes, it is great for the therapeutic potential of hypoinsulinism class patients with type Ⅰ DM.
The present invention proposes a kind of hypoglycemic purposes of beans taro leaf polyose, is that natural hypoglycemic is developed in the fields such as medicine, Food Science
Product provides new resources.It is related to application of the beans taro leaf polyose in preparing antihypelipidemic product simultaneously.That listed above is only the present invention
Implementation several examples.It is clear that the invention is not restricted to above example, acceptable there are many deformations.The common skill of this field
All deformations that art personnel directly can export or associate from present disclosure are considered as the protection model of the present invention
It encloses.
Claims (4)
1. the hypoglycemic purposes of beans taro leaf polyose, it is characterised in that:
It is used to prepare antihypelipidemic foodstuff and/or drug.
2. the hypoglycemic purposes of beans taro leaf polyose according to claim 1, it is characterised in that:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced HepG2 cells to glucose consumption ability.
3. the hypoglycemic purposes of beans taro leaf polyose according to claim 1, it is characterised in that:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced HepG2 cells to glucose uptake ability.
4. the hypoglycemic purposes of beans taro leaf polyose according to claim 1, it is characterised in that:
It is used to prepare the antihypelipidemic foodstuff and/or drug for improving high sugar induced RINm5F cells synthesis excreting insulin ability.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109512812A (en) * | 2018-12-14 | 2019-03-26 | 江苏省农业科学院 | A kind of glucose induction human liver cancer cell HepG2 establishes the method and application of diabetes model |
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2018
- 2018-04-28 CN CN201810404020.3A patent/CN108685938A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 安家炜等: "美国豆芋地上部位有效成分及α- 葡萄糖苷酶抑制活性研究", 《核农学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109512812A (en) * | 2018-12-14 | 2019-03-26 | 江苏省农业科学院 | A kind of glucose induction human liver cancer cell HepG2 establishes the method and application of diabetes model |
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