CN110448642A - A kind of rhizoma polygonati fermentation material, preparation method and application - Google Patents
A kind of rhizoma polygonati fermentation material, preparation method and application Download PDFInfo
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- CN110448642A CN110448642A CN201910817897.XA CN201910817897A CN110448642A CN 110448642 A CN110448642 A CN 110448642A CN 201910817897 A CN201910817897 A CN 201910817897A CN 110448642 A CN110448642 A CN 110448642A
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- rhizoma polygonati
- fermentation material
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- slurries
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229940100389 Sulfonylurea Drugs 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- QWCKQJZIFLGMSD-UHFFFAOYSA-N alpha-aminobutyric acid Chemical class CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 150000004283 biguanides Chemical class 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 208000033679 diabetic kidney disease Diseases 0.000 description 1
- 238000013118 diabetic mouse model Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
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- 201000010063 epididymitis Diseases 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000004426 flaxseed Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
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- 238000010438 heat treatment Methods 0.000 description 1
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- 235000008216 herbs Nutrition 0.000 description 1
- 235000009200 high fat diet Nutrition 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
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- 238000005286 illumination Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
- 208000006443 lactic acidosis Diseases 0.000 description 1
- 229940078795 lactoferrin Drugs 0.000 description 1
- 235000021242 lactoferrin Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 229940010454 licorice Drugs 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 230000000291 postprandial effect Effects 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- YROXIXLRRCOBKF-UHFFFAOYSA-N sulfonylurea Chemical class OC(=N)N=S(=O)=O YROXIXLRRCOBKF-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- BSYVTEYKTMYBMK-UHFFFAOYSA-N tetrahydrofurfuryl alcohol Chemical group OCC1CCCO1 BSYVTEYKTMYBMK-UHFFFAOYSA-N 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/40—Shaping or working of foodstuffs characterised by the products free-flowing powder or instant powder, i.e. powder which is reconstituted rapidly when liquid is added
-
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8969—Polygonatum (Solomon's seal)
-
- 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
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/121—Brevis
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention discloses a kind of rhizoma polygonati fermentation materials, preparation method and application, belong to biological medicine manufacturing technology field.The preparation method selects lactic acid bacteria class probiotics strain as leavening, and leavening is seeded in the rhizoma polygonati slurries after sterilizing and is fermented, rhizoma polygonati fermentation material is obtained；Wherein leavening is mixed to prepare by lactobacillus plantarum and Lactobacillus brevis according to bacteria suspension volume ratio 1:0-10, and leavening is accessed in the rhizoma polygonati slurries after sterilizing by the inoculum concentration of 1%-30%, and the 10-80h that ferments at 20-36.5 DEG C；The invention also discloses the applications of above-mentioned rhizoma polygonati fermentation material and above-mentioned rhizoma polygonati fermentation material.Preparation method of the invention considerably reduces rhizoma polygonati small molecular sugar substance, it is fermented using the lactic acid bacteria strains of high yield aminobutyric acid to rhizoma polygonati, not only the small molecule carbohydrate in rhizoma polygonati had been eliminated, but also has remained the hypoglycemic related activity ingredients such as saponin(e and Siberian solomonseal rhizome polysaccharide in rhizoma polygonati；The ingredients such as the aminobutyric acid generated that ferment are cooperateed with the active constituent in rhizoma polygonati generates anti-diabetes B effect.
Description
Technical field
The present invention relates to biological medicine manufacturing technology fields, and in particular to a kind of rhizoma polygonati fermentation material, preparation method and application.
Background technique
Diabetes are current one of the major diseases for threatening human health.Diabetes be after cardiovascular and cerebrovascular disease and tumour it
The third-largest disease of human health is endangered afterwards, and complication includes hyperlipidemia, diabetic nephropathy, diabetic eye diseases, hepatic injury, the heart
Vascular diseases etc..Treat the needs long-time medication such as first-line drug such as sulfonylurea, biguanides of diabetes, consumption cost compared with
Height, while long-time medication easily causes the toxic side effects such as hypoglycemia, lactic acidosis, abdominal distension, diarrhea, therefore, exploitation is new
Type, antidiabetic medicine efficiently, less toxic or adjuvant treatment are still the hot spot of biomedicine field research with product.
Traditional Chinese medicine has long-term clinical practice experience in terms for the treatment of diabetes, it was found that many has the medicine of significant curative effect
Object.Wherein rhizoma polygonati is exactly one of the Chinese medicine for having significant curative effect to diabetes.It is that liliaceous plant Yunnan is yellow that China's pharmacopeia, which records rhizoma polygonati,
Smart Polygonatum kingianum Coll.et Hemsl., rhizoma polygonati Polygonatum sibiricum Red. spend more Huang
The dry rhizome of smart Polygonatum cyrtonema Hua.First recorded in " Mingyi Bielu ", effect are as follows: boosting qi and nourishing yin, invigorating the spleen,
Moistening lung, kidney-nourishing.There are many record using rhizoma polygonati relevant prescription treatment diabetes, such as 2015 version " Chinese Pharmacopoeia " record it is thirsty
Happy Yiganning capsule, ginseng the essence ball that quenches the thirst are used to the treatments of diabetes, also using rhizoma polygonati as one of primary raw material in side.Also there is a large amount of text
Offering report confirms that Siberian solomonseal rhizome polysaccharide, rhizoma polygonati total extract have preferable hypoglycemic effect.
Rhizoma polygonati defends integration of drinking and medicinal herbs raw material variety as defined in planning commission as country, itself has the work of certain alleviation diabetes
With, but since the small molecule carbohydrate in rhizoma polygonati such as sucrose, glucose and fructose content are higher, it is measured according to us, in dry rhizoma polygonati
Small molecule contents of saccharide mostly 10% or so, the rhizoma polygonati small molecular sugar contents that are produced from individual places are more than 20%, due to existing
Small molecular sugar content in technology rhizoma polygonati is high, therefore is not suitable for patients with diabetes mellitus.
Summary of the invention
The defect that the purpose of the invention is to overcome rhizoma polygonati to use as hypoglycemic drug, is removed by microbe fermentation method
The small molecule carbohydrate in rhizoma polygonati is removed, to improve response to treatment of the rhizoma polygonati for diabetes.
There is provided a kind of preparation method of rhizoma polygonati fermentation material, selections can generate aminobutyric acid for an object of the present invention
Leavening is seeded in the rhizoma polygonati slurries after sterilizing and ferments, obtain rhizoma polygonati fermentation material as leavening by lactic acid bacteria class probiotics.
Preferably, leavening is mixed to prepare by lactobacillus plantarum and Lactobacillus brevis according to bacteria suspension volume ratio 1:0-10.
Preferably, leavening is accessed in the rhizoma polygonati slurries after sterilizing by the inoculum concentration of percent by volume 1%-30%, and
Ferment 10-80h at 20-36.5 DEG C.
Preferably, the pH of rhizoma polygonati slurries is 6.0 ± 2, when fermentation liquid pH is reduced to 3.0-3.5 to indicate that fermentation is completed.
Preferably, rhizoma polygonati slurries the preparation method is as follows: fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, crush, add water, obtain
Rhizoma polygonati slurries；The sterilized processing of rhizoma polygonati slurries, the rhizoma polygonati slurries after must sterilizing.
Preferably, rhizoma polygonati dry weight in rhizoma polygonati slurries: the mass ratio of water is 1:3~10.
Preferably, rhizoma polygonati fermentation material is freeze-dried under vacuum conditions after quick prefreezing, crush, obtains rhizoma polygonati fermentation
Object freeze-dried powder.
Preferably, the concentration of bacteria suspension is 1 × 106-1×108A/mL.
The second object of the present invention is to provide the preparation method of above-mentioned rhizoma polygonati fermentation material preparation-obtained rhizoma polygonati fermentation material；
The third object of the present invention is to provide rhizoma polygonati fermentation material in food, drug or the health of preparation treatment diabetes B
Application in product.
Compared with prior art, the beneficial effects of the present invention are:
The present invention considerably reduces rhizoma polygonati small molecular sugar substance by fermentation method, and the rhizoma polygonati fermentation material being prepared is small
Molecule sugared content is substantially reduced, and is more suitable for type 2 diabetic patient and is taken.
The present invention ferments to rhizoma polygonati using the lactic acid bacteria strains of high yield aminobutyric acid, can both effectively remove in rhizoma polygonati
Small molecule carbohydrate, and the hypoglycemics related activity ingredient such as can retain saponin(e and Siberian solomonseal rhizome polysaccharide in rhizoma polygonati；And high yield amino fourth
The ingredients such as aminobutyric acid that the lactic acid bacteria of acid generates also have some improvement the effect of diabetic symptom, can be with the activity in rhizoma polygonati
Ingredient collaboration generates anti-diabetes B effect.
Rhizoma polygonati fermentation material freeze-dried powder prepared by the present invention can be applied to the preparation treatment food of diabetes B, drug or
In health-oriented products, such as rhizoma polygonati fermentation material freeze-dried powder can be made as meal replacement powder, food, pharmaceutical raw material and be used to treat diabetes.
Detailed description of the invention
Fig. 1 is Siberian solomonseal rhizome polysaccharide of the present invention and rhizoma polygonati fermentation material polysaccharide infrared chromatogram；
Fig. 2 is influence of the PS of the present invention and FPS to diabetes B mouse fasting blood-glucose；
Fig. 3 is influence of the PS of the present invention and FPS to diabetes B mouse Fasting insulin level；
Fig. 4 is the influence that PS of the present invention and FPS resists index to diabetes B mouse islets element；
Fig. 5 is influence of the PS of the present invention and FPS to diabetes B mice serum glycosylated hemoglobin.
Specific embodiment
The present invention is made combined with specific embodiments below and further being elaborated, the embodiment is served only for explaining this
Invention, is not intended to limit the scope of the present invention.Test method as used in the following examples is normal unless otherwise specified
Rule method；Used material, reagent etc., unless otherwise specified, for the reagent and material commercially obtained.
Embodiment one:
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:4 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
By cultured lactobacillus plantarum CICC 24202 (Lactobacillus plantarum, abbreviation LP), short cream bar
Bacterium YM 1301 (Lactobacillus breris, abbreviation LB)；
Bacteria suspension is that 1:1 is mixed to prepare leavening with volume ratio, is accessed in rhizoma polygonati slurries by 10% inoculum concentration, 36.5
It ferments for 24 hours at DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain freeze-dried powder.
(5) rhizoma polygonati fermentation material freeze-dried powder can be packed directly as having the generation meal product for reducing blood glucose effect, everyone is every
Day amount is 5~30g or so.
The microorganism that the fermentation process of the present embodiment uses is that country defends the strain list that can be used for food as defined in planning commission
In include strain, selected strain during the fermentation, can quickly using the small molecule carbohydrate contained in rhizoma polygonati powder, with
Achieve the purpose that reduce the small molecule carbohydrate for being not suitable for patients with diabetes mellitus contained in rhizoma polygonati；In addition, selected strain
Also some functional components be can produce, synergistic effect, sanatory effect are generated.
Embodiment two:
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:10 plus water, adjust pH to 6.2.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
LP bacterial strain, LB bacterial strain bacteria suspension are mixed to prepare leavening with volume ratio for 1:1, accessed by 10% inoculum concentration yellow
In refining, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain freeze-dried powder.
(5) rhizoma polygonati fermentation material freeze-dried powder is mixed with the pretreatment of other auxiliary materials, according to conventional beverage production technology, sized mixing,
Stabilization processes, seasoning, homogeneous, it is filling, sterilizing, finished product process, be fabricated to beverage.It is mixed in proportion after pretreatment of raw material,
It is dissolved, is stirred evenly with water or fruit juice, honey etc.；Then appropriate emulsion stabilizer processing is added, is sufficiently stirred, adds it
His flavoring substance, forms specific flavor, then homogeneous, is formed than more uniform, stable colloidal solution；It is last filling, it goes out
Finished product is packaged as after bacterium, as having the beverage for reducing blood glucose effect to use, the amount of drinking is lyophilized by rhizoma polygonati fermentation material daily for everyone
Powder is calculated as 5~30g or so.
Embodiment three:
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:5 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
LP bacterial strain, LB bacterial strain bacteria suspension are mixed to prepare leavening with volume ratio for 1:1, accessed by 10% inoculum concentration yellow
In refining, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain freeze-dried powder.
(5) rhizoma polygonati fermentation material freeze-dried powder and raw-food material such as Rhizoma Polygonati extract, licorice, linseed meal etc. mix,
And conventional flavoring substance is added, it is used after mixing as with the multi-functional meal replacement powder of hypoglycemic effect, everyone amount of drinking daily
5~30g or so is calculated as by rhizoma polygonati fermentation material freeze-dried powder.
Example IV:
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:4 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
LP bacterial strain, LB bacterial strain bacteria suspension are mixed to prepare leavening with volume ratio for 1:1, accessed by 10% inoculum concentration yellow
In refining, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain rhizoma polygonati fermentation material freeze-dried powder.
(5) by prepare containing active constituent be rhizoma polygonati fermentation material be lyophilized powder chewable tablet 1000 for raw materials used and auxiliary material and
Its quality proportioning is as follows: rhizoma polygonati fermentation material freeze-dried powder 1435g, dextrin 45g, magnesium stearate 5g, Aspartame 15g, by galenic pharmacy
The common process of tablet carries out, every slice weight 1.5g, the every freeze-dried powder of fermentation material containing rhizoma polygonati 1.43g.Usage and dosage: it is oral, every time 3
~6, three to five times on the one, the amount of drinking by rhizoma polygonati fermentation material freeze-dried powder is calculated as 5~30g or so daily for everyone.
Embodiment five:
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:4 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
LP bacterial strain, LB bacterial strain bacteria suspension are mixed to prepare leavening with volume ratio for 1:1, accessed by 10% inoculum concentration yellow
In refining, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain freeze-dried powder.
(5) by prepare be rhizoma polygonati rhizoma polygonati fermentation material freeze-dried powder object plane packet containing active constituent for raw materials used and auxiliary material and its
Quality proportioning is as follows: rhizoma polygonati fermentation material freeze-dried powder 80g, cane sugar powder 850g flour, vegetable oil 50g, active dry yeasr 20g.According to normal
It advises breadmaking process and makes bread.Bread 250g, the freeze-dried powder of fermentation material containing rhizoma polygonati 20g are eaten for each person every day.
In the food such as such as biscuit, cake, rhizoma polygonati fermentation material freeze-dried powder can also be added, by rhizoma polygonati fermentation material for each person every day
5~30g of amount of freeze-dried powder is advisable.
Embodiment six
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:4 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are sterilized 30min with high-pressure sterilizing pot at 121 DEG C.
(3) inoculation fermentation:
By LP bacterial strain by 10% inoculum concentration access rhizoma polygonati slurry, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain rhizoma polygonati fermentation material freeze-dried powder.
(5) by prepare containing active constituent be rhizoma polygonati fermentation material be lyophilized powder chewable tablet 1000 for raw materials used and auxiliary material and
Its quality proportioning is as follows: rhizoma polygonati fermentation material freeze-dried powder 1435g, dextrin 45g, magnesium stearate 5g, Aspartame 15g, by galenic pharmacy
The common process of tablet carries out, every slice weight 1.5g, the every freeze-dried powder of fermentation material containing rhizoma polygonati 1.43g.Usage and dosage: it is oral, every time 3
~6, three to five times on the one, the amount of drinking by rhizoma polygonati fermentation material freeze-dried powder is calculated as 5~30g or so daily for everyone.
Embodiment seven
(1) raw material pre-treatment:
Fresh rhizoma polygonati or dry rhizoma polygonati are cleaned, the part that removal goes mouldy and damages by worms is crushed, rhizoma polygonati dry weight: water
Rhizoma polygonati slurries are obtained for 1:4 plus water, adjust pH to 6.5.
(2) it sterilizes:
Rhizoma polygonati slurries are evenly laid out on microwave oven internal pallet according to thickness 5cm, and sterilization time 240s, sterilize function
Rate is 6 000W.
(3) inoculation fermentation:
LP bacterial strain, LB bacterial strain bacteria suspension are mixed to prepare leavening with volume ratio for 1:1, accessed by 10% inoculum concentration yellow
In refining, ferment for 24 hours at 36.5 DEG C.
(4) it is freeze-dried:
It is dry in the freeze drier that vacuum degree is 0.05~0.1MPa by rhizoma polygonati tunning after -20 DEG C of quick freezings
It is dry to crush afterwards for 24 hours, obtain freeze-dried powder.
(5) by prepare be rhizoma polygonati rhizoma polygonati fermentation material freeze-dried powder object plane packet containing active constituent for raw materials used and auxiliary material and its
Quality proportioning is as follows: rhizoma polygonati fermentation material freeze-dried powder 80g, cane sugar powder 850g flour, vegetable oil 50g, active dry yeasr 20g.According to normal
It advises breadmaking process and makes bread.Bread 250g, the freeze-dried powder of fermentation material containing rhizoma polygonati 20g are eaten for each person every day.
In the food such as such as biscuit, cake, rhizoma polygonati fermentation material freeze-dried powder can also be added, by rhizoma polygonati fermentation material for each person every day
5~30g of amount of freeze-dried powder is advisable.
We have detected rhizoma polygonati fermentation front and back small molecule carbohydrate, total starches and rhizoma polygonati fermentation material freeze-dried powder aminobutyric acid
Changes of contents, it was demonstrated that after fermentation process, small molecule contents of saccharide in rhizoma polygonati fermentation material freeze-dried powder is significantly reduced, function because
Sub- Gamma-propalanine content significantly improves.It is bright that pharmacodynamic experiment confirms that rhizoma polygonati fermentation material freeze-dried powder has diabetes B related symptoms
Aobvious improvement result.Related experiment is as follows:
The Changeement of related main component after one rhizoma polygonati fermentation process
1 materials and methods
1.1 experimental material
Rhizoma polygonati is purchased from Shangnan County, is identified as the rhizome of liliaceous plant rhizoma polygonati (P.sibiricum Red.).Test is used
Bacterial strain is lactobacillus plantarum (Lactobacillus plantarum, abbreviation LP) CICC 24202, is purchased from Chinese industrial microorganism
Culture presevation administrative center；Lactobacillus brevis (Lactobacillus breris, abbreviation LB) YM 1301, it is micro- to be stored in Yunnan Province
Biological study institute.
Other chemical reagent are commercially available general reagent.
1.2 experimental method
1.2.1 the preparation of rhizoma polygonati fermentation material freeze-dried powder
Water is added with the ratio of solid-liquid ratio 1:4 (m/V), after mixing up to rhizoma polygonati in the dry once purged crushing of rhizoma polygonati
Slurries adjust its pH to 6.0 ± 0.5, are sterilized at 121 DEG C 20min after packing with high-pressure steam sterilizing pan；Rhizoma polygonati after sterilizing
In the bacteria suspension of the ratio access 10% of LP:LB=1:1 (V/V), (adjustment bacteria suspension concentration is 1 × 10 to slurries8A/mL.), it mixes
In 36 DEG C of fermentation 60h after even；Fermentation rhizoma polygonati is starched after -20 DEG C of quick freezings, in the freezing that vacuum degree is 0.05~0.1MPa
Drying machine drying crushes afterwards for 24 hours, obtains rhizoma polygonati fermentation material freeze-dried powder.
1.2.2 the measurement of glucose content
Glucose oxidase-peroxidase method, Glucose estimation kit (the limited public affairs of Shanghai Rong Sheng biology medicine company
Department), it is operated according to kit specification.Rhizoma polygonati powder, each 500.0mg of rhizoma polygonati fermentation material freeze-dried powder are accurately weighed respectively, it is molten respectively
In 10.0mL distilled water, centrifuging and taking supernatant is as sample to be tested.
1.2.3 total reducing sugars assay
Total reducing sugars assay with 3,5- dinitrosalicylic acid (DNS) colorimetric method, with glucose as a standard mark by product, production
Directrix curve；Accurately weigh rhizoma polygonati powder respectively, rhizoma polygonati fermentation material freeze-dried powder 50.0mg is dissolved in 100mL distilled water, centrifugation, take
Clearly, 1.0mL supernatant is respectively taken, 2.0mL DNS reagent is separately added into, oscillation mixes, 90 DEG C of heating water bath 5min, then in room temperature
Cooling 20min, then add 2.0mL distilled water to mend volume to 4.0mL into every test tube, it mixes.Measure the extinction at 520nm
Value.Concentration of reduced sugar in sample to be tested is calculated according to standard curve.
1.2.4 determination of polysaccharide
Determination of polysaccharide uses phend-sulphuric acid in rhizoma polygonati fermentation material freeze-dried powder.Accurately weigh the dry Huang to constant weight
Fine powder end and each 1.00g of rhizoma polygonati fermentation material freeze-dried powder are separately added into the distilled water of 20 times of amounts, in boiling water bath in 50mL conical flask
Middle heating extraction 2h is repeated to extract 3 times, after combined extract, extracting solution is concentrated into the 1/10 of original volume, is added into filtrate
95% ethyl alcohol of 4 times of volumes stands 12h, and centrifugation discards supernatant, and underclad portion uses alcohol precipitation 2 times repeatedly of 95% ethyl alcohol again, after centrifugation
Gained precipitating is saccharide portion after freeze-drying；Each 50mg of above-mentioned polysaccharide is accurately weighed, is settled to after being dissolved with distilled water
500mL is made into the prepare liquid of 0.1mg/mL.Accurate 1.0mL the sample solution to be tested of drawing in 3 10mL tool plug test tubes, it is backward respectively
Add 5% phenol solution of 1.0mL in group, gently oscillation mixes, and is then slowly added to the 5.0mL concentrated sulfuric acid into each group again, sufficiently shakes
Mixing is swung, after 90 DEG C of water-bath 20min, the cooling 30min of room temperature, measures the light absorption value at 490nm wavelength.With glucose as a standard
Product make standard curve, calculate polyoses content in sample.
1.2.5 the infrared chromatography of polysaccharide analyzes (IR)
The same 1.2.4 of the extracting method of polysaccharide (extraction of saccharide portion)
By the Siberian solomonseal rhizome polysaccharide of Siberian solomonseal rhizome polysaccharide and rhizoma polygonati fermentation material freeze-dried powder respectively from KBr in the ratio of 1:100 be placed in through
In the dried and clean agate mortar that dehydrated alcohol is cleaned, the two is mixed, is firmly ground to fine-powdered with grinding hammer, it will be thin in right amount
Pulverulent mixture is placed in compression mold clean after alcohol is cleaned, and transparent sample thin slice is made in pressurization 1min or so.By what is pressed
Complete transparent sample thin slice is placed on infrared chromatograph specimen holder, in 4000-400cm-1Scanning, first surveys blank background in range, then
The IR of PSP, FPSP are surveyed, and records result.
1.2.6 the measurement of aminobutyric acid -- HPLC method
Chromatographic condition is as follows: DiamonsilC18 chromatographic column, and 30 DEG C of column temperature, specification is 250mm × 4.6mm, ultraviolet detection
Wavelength 254nm, 20 μ L of sample volume.Mobile phase A is methanol, and Mobile phase B is tetrahydrofuran-methanol -0.05mol/L sodium acetate
(pH6.2), ratio is (5:75:420), flow velocity 0.8mL/min, gradient elution.
Sample preparation methods: weighing rhizoma polygonati powder and rhizoma polygonati fermentation material freeze-dried powder 1.00g respectively, and 100mL distilled water is added, and surpasses
Sound extracts 30min, filtering.100 μ L of filtrate is taken, borate buffer 1mL and the o-phthalaldehyde (OPA) that 0.4mol/L is added are derivative
200 μ L of liquid, after mixing, derivative reaction 30min takes 10 μ L sample introductions to measure.
Standard curve making: using aminobutyric acid (GABA) concentration of standard solution as abscissa, pass through high performance liquid chromatography
Measuring peak area is ordinate, draws standard curve, establishes regression equation.
2 experimental results
2.1 glucose content
The variation (n=3) of glucose content in the rhizoma polygonati of table 1-1 fermentation front and back
Note: alphabetical A, B expression is examined through t between the two, and difference has extremely significant meaning (p < 0.01) table 1-1 it is found that rhizoma polygonati
The content of middle glucose is had decreased to 0.39% after fermentation by 1.04% before fermenting, and data difference has significant meaning, the number
According to consistent with the result predicted before, illustrate that fermentation energy reduces the content of glucose in rhizoma polygonati.
2.2 total reducing sugars contents
It is computed, glucose standard curve y=2.6434x+0.0048, R2=0.9973；It is calculated using standard curve
Content of reducing sugar in sample, as a result such as table 1-2.By table 1-2 it is found that under also there is significantly the content of reduced sugar before and after fermentation
Drop, has decreased to 3.83% from 9.47%, data difference has significant meaning；Reduced sugar be mainly by with reproducibility monosaccharide and
The small molecules carbohydrate such as disaccharides such as glucose, fructose, galactolipin, lactose, maltose composition, polysaccharide do not have reproducibility generally.Also
The decline of originality sugared content illustrates the small molecule contents of saccharide decline after fermenting in rhizoma polygonati.
The variation (n=3) of content of reducing sugar in the rhizoma polygonati of table 1-2 fermentation front and back
Note: alphabetical A, B expression is examined through t between the two, and difference has extremely significant meaning (p < 0.01)
2.3 polyoses content
It is computed, with glucose as a standard product, the equation using phend-sulphuric acid production standard curve is y=6.1811x
+ 0.0443, R2=0.9975；Sample measurement substitutes into standard curve calculated result as shown in table 1-3, as can be seen from the table,
Siberian solomonseal rhizome polysaccharide content after fermentation is higher than the content for the preceding Siberian solomonseal rhizome polysaccharide that ferments, and difference has significant meaning (p < 0.01).Rhizoma polygonati is more
For sugar as one of most important active constituent of rhizoma polygonati, having a large amount of document proves that Siberian solomonseal rhizome polysaccharide has hypoglycemic, reducing blood lipid etc.
Effect, the raising of the Siberian solomonseal rhizome polysaccharide content after fermentation may be the consumption due to small molecule carbohydrate and make the relative amount of polysaccharide
It improves, it is also possible to be that bacterial strain produces some polysaccharide in the metabolic process in fermentation process.
The variation of table 1-3 fermentation front and back Polysaccharide in Polygonatum sibiricum
Note: alphabetical A, B expression is examined through t between the two, and difference has extremely significant meaning (p < 0.01)
The analysis of 2.4 infrared chromatographies
The infrared spectroscopy of Siberian solomonseal rhizome polysaccharide (PSP) and rhizoma polygonati fermentation material freeze-dried powder Siberian solomonseal rhizome polysaccharide (FPSP) (wave as shown in Figure 1:
Number 4000cm-1-400cm-1 middle infrared), PSP and FPSP are in 3429cm-1、1635cm-1、1394cm-1、1065cm-1Place has altogether
Same characteristic peak, these peaks are the characteristic absorption peaks of polysaccharide.In 3429cm-1The absorption peak at place is the feature of O-H stretching vibration
Peak；And in 1635cm-1The absorption peak of the moderate strength at place is as caused by C=O stretching vibration, and prompting may in two kinds of polysaccharide
There are also uronic acids；Absorption peak at 1394cm-1 is the characteristic peak of C-H bending vibration；Absorption peak at 1065cm-1 is polysaccharide
Characteristic absorption peak.
The infrared spectrogram of two kinds of polysaccharide is compared, it can be found that the rhizoma polygonati after Siberian solomonseal rhizome polysaccharide (PSP) and fermentation is more
Sugared (FPSP) absorption peak similarity is higher, shows that the variation of fermentation front and back polysaccharide is smaller.Finger at 1600cm-1~1000cm-1
There is subtle difference at line peak, it may be possible to which lactic acid bacteria metabolism produces caused by some new substances in fermentation process.
The content of 2.5 aminobutyric acids
According to 1.2.6 method, using GABA concentration of standard solution as abscissa, peak area is measured by high performance liquid chromatography
For ordinate, standard curve is drawn, establishes regression equation, i.e., are as follows: y=0.0803x-2.0728, R2=0.9992.Measure sample
Corresponding peak area substitute into calibration curve equation calculate, obtain the content such as following table of aminobutyric acid in sample.It confirms through above-mentioned two
After kind strain fermentation, active constituent aminobutyric acid is contained in rhizoma polygonati fermentation material freeze-dried powder.
The variation of Gamma-propalanine content in the rhizoma polygonati of table 1-4 fermentation front and back
3 experiment conclusions
Based on the above results as can be seen that during rhizoma polygonati fermentation, lactobacillus plantarum (LP) and Lactobacillus brevis (LB)
The glucose and other small molecule contents of saccharide in rhizoma polygonati are effectively reduced, while relevant to its hypoglycemic effect in rhizoma polygonati
Active material such as polysaccharide component content has small size raising, and polysaccharide relative amount improves after fermentation process, so after fermentation process
Rhizoma polygonati be more suitable for patients with diabetes mellitus.Since the aminobutyric acid substance generated in fermentation process is to diabetic's correlation
Symptom also has preferable improvement effect, so the rhizoma polygonati after fermentation theoretically should have better response to treatment to diabetes.
Two, rhizoma polygonati fermentation material freeze-dried powder improves the pharmacodynamic study of diabetes B
2.1 materials and methods
2.1.1 experimental material
Rhizoma polygonati is purchased from Shangnan County, is identified as the rhizome of liliaceous plant rhizoma polygonati (P.sibiricum Red.).Rhizoma polygonati hair
Ferment object freeze-dried powder prepares same preceding method, and water is added with the ratio of solid-liquid ratio 1:4 (m/V) in the dry once purged crushing of rhizoma polygonati,
It is starched after mixing up to rhizoma polygonati, adjusts its pH to 6.0 ± 0.5, sterilized at 121 DEG C after packing with high-pressure steam sterilizing pan
20min；After sterilizing rhizoma polygonati slurry by LP:LB=1:1 (V/V) ratio access 10% bacteria suspension (adjustment bacteria suspension concentration be 1
×108A/mL), in 37 DEG C of fermentation 60h after mixing thoroughly；Fermentation rhizoma polygonati is starched after -20 DEG C of quick freezings, is 0.05 in vacuum degree
The freeze drier drying of~0.1MPa crushes afterwards for 24 hours, obtains rhizoma polygonati fermentation material freeze-dried powder.
Main agents: streptozotocin (STZ) (Sigama company, C3H15N3O7), Metformin hydrochloride (Shandong Ming Ren good fortune
Rui Da object Pharmacy stock Co., Ltd), mouse islets element (INS) ELISA detection kit (Shanghai enzyme is far biological), mouse saccharification
Hemoglobin (GHb) ELISA detection kit (Shanghai enzyme is far biological), mouse glucagon-like peptide 1 (GLP-1) ELISA inspection
Test agent box (Shanghai enzyme is far biological), total cholesterol (T-CHO) testing cassete (Bioengineering Research Institute is built up in Nanjing), triglycerides
(TG) testing cassete (Bioengineering Research Institute is built up in Nanjing), (life is built up in Nanjing to low density lipoprotein cholesterol (LDL-C) testing cassete
Object Graduate School of Engineering), high-density lipoprotein cholesterol (HDL-C) testing cassete (Bioengineering Research Institute is built up in Nanjing).
Other chemical reagent are commercially available general reagent.
2.1.2 experimental animal
Male mouse of kunming, 18g ± 2g are purchased from Xi'an Communications University's medical board Experimental Animal Center, animal productiong license
Card SCXK (Shan) 2017-003.
2.1.3 experimental animal grouping and processing
It male mouse of kunming 100, weight 18g ± 2g, raises in animal house, clean grade is 22 ± 1 DEG C of constant temperature, relatively wet
40%-70% is spent, the experiment feed and free water of standard are given in 12h illumination, and adaptive feeding starts to test after a week.It is dynamic
The grouping of object and processing method are as shown in table 2-1: mouse is randomly divided into 2 groups: Normal group (Normal Control,
N.control) and modeling group, Normal group (10) give normal diet, and modeling group (90) gives high lipid food (High
Fat Diet, HFD).Joint streptozotocin (STZ) intraperitoneal injection, which is fed, by high lipid food destroys islet cells 2 types of induction
Diabetic mouse model.After high lipid food is fed 4 weeks, it is deprived of food but not water 12h, (STZ is dissolved in intraperitoneal injection 70mg/kg STZ
0.1mol/L citric acid-sodium citrate buffer solution, pH:4.4 are configured to the solution that concentration is 1%, ready-to-use.Normal control
The citric acid-sodium citrate buffer solution of group injection respective volume), STZ is injected again with identical dosage after 72h, is infused for second
It penetrates after STZ 72h and measures fasting blood-glucose, fasting blood-glucose > 11.1mmol/L and after 3 weeks fasting blood-glucose still > with blood glucose meter
The mouse of 11.1mmol/L thinks the successful diabetic mice of modeling.
The successful mouse of modeling is randomly divided into 6 groups: model group (Model), positive drug group (Positive Control,
P.control), rhizoma polygonati group (Polygonatum Sibiricum, PS), rhizoma polygonati+LP and LB Mixed Microbes freeze-dried powder group (PS+LP+
It after LB, LP and LB are cultivated in MRS fluid nutrient medium respectively, is freeze-dried after the mixing of bacteria suspension equal proportion, preparation lactic acid bacteria is frozen
Dry powder, by experiment one in 2.5 methods measurement lactic acid bacteria freeze drying powder in Gamma-propalanine content after, by lactic acid bacteria freeze drying powder with do not send out
Ferment rhizoma polygonati powder mixing, so that the Gamma-propalanine content in mixture is suitable with content in rhizoma polygonati fermentation material freeze-dried powder.) rhizoma polygonati fermentation
Object freeze-dried powder low dose group (Low Dose of Fermented Polygonatum Sibiricum, FPSL), rhizoma polygonati fermentation material
Freeze-dried powder high dose group (High Dose of Fermented Polygonatum Sibiricum, FPSH).
N.control group: normal diet, normal diet drinking-water, the physiological saline of daily stomach-filling equal volume are given；
Model group: high lipid food, normal diet drinking-water, the physiological saline of daily stomach-filling equal volume are given；
P.control group: high lipid food, normal diet drinking-water, daily stomach-filling Metformin hydrochloride 150mg/kg are given；
PS group: word material high in fat, normal diet drinking-water, daily stomach-filling rhizoma polygonati powder 0.5g/kg are given；
PS+LB+LP group: word material high in fat, normal diet drinking-water, daily stomach-filling rhizoma polygonati+LP and LB Mixed Microbes freeze-dried powder are given
0.5g/kg；
FPSL group: word material high in fat, normal diet drinking-water, daily stomach-filling rhizoma polygonati fermentation material freeze-dried powder freeze-dried powder 0.25g/ are given
kg；
FPSH group: word material high in fat, normal diet drinking-water, daily stomach-filling rhizoma polygonati fermentation material freeze-dried powder freeze-dried powder 0.5g/ are given
kg。
The grouping of table 2-1 experimental animal and administrations
During experiment, the daily active degree for observing animal, hair color, smooth degree, state of mind etc., and monitor mouse
Diet amount of drinking water, urine volume and weight.Successive administration 4 weeks, mouse was deprived of food but not water 12h after the last administration, and every mouse weighing is simultaneously
Blood is taken in orbital venous plexus after measurement fasting blood-glucose, centrifuging and taking serum saves after packing in -20 DEG C, is used for subsequent measurements.
2.2 experiment test indexs and method
2.2.1 the weight of animals detects
Mouse after being deprived of food but not water 12h, weighs before administration and after administration and records its weight.
2.2.2 fasting blood-glucose detects
Mouse after being deprived of food but not water 12h, surveys its fasting blood sugar respectively at administration front and back.
2.2.3 oral glucose tolerance measures
Mouse puts to death preceding 1 week row oral glucose tolerance test (oral glucose tolerance test, OGTT).
It is overnight before test to be deprived of food but not water 12h, glucose is given with the dosage stomach-filling of 2g/kg.After blood glucose blood sampling point is empty stomach, load
30min, 60min and 120min measure tail vein blood glucose value with three promise stabilizing blood sugar instrument.The blood glucose value of various time points is denoted as
G0、G30、G60、G120, glucose tolerance curve figure, and area (area under the under calculated curve are drawn according to testing result
Curve, AUC).Calculation formula are as follows: AUC (mmol/L)=(1/2 × G0+G30+G60+1/2×G120)/2。
2.2.4 insulin tolerance detects
Mouse puts to death preceding row insulin tolerance test in 3 days (insulin tolerance test, ITT).Fasting before testing
2h, measurement 0min tail vein blood glucose value and immediately intraperitoneal injection 0.75IU/kg insulin, 30min, 60min after injection,
120min measures tail vein blood glucose value respectively, and the blood glucose value of various time points is denoted as G0、G30、G60、G120, according to testing result
Draw insulin tolerance curve graph, and area (AUC) under calculated curve.Calculation formula are as follows: AUC (mmol/L)=(1/2 × G0+
G30+G60+1/2×G120)/2。
2.2.5 Fasting insulin level measures
Illustrate to operate according to kit, detects the content of mice serum Fasting insulin level.Insulin is calculated according to following formula
Resist index:
HOMA-IR=fasting blood-glucose (FPG) × fasting insulin (FINS)/22.5
2.2.6 serum glycated hemoglobin measures
Illustrate to operate according to corresponding kit, detects the content of mice serum glycosylated hemoglobin.
2.2.7 lipid determination
Total cholesterol (Total Cholesterol, T-CHO) in serum, total triglycerides (Triglyceride, TG),
The measurement of low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) content illustrates according to kit
It carries out.
2.2.8 the measurement of glucagon-like peptide 1
Illustrate to operate according to kit, detects the content of glucagon-like peptide 1 (GLP-1) in mice serum.
2.2.9 data statistics processing method
Using 5 analyzing and processing data of software SPSS 16.0 and GraphPad Prism, with average value ± standard error (Mean
± SEM) mode express data.The analysis of significant difference then uses t to examine between group, and P < 0.05 indicates that statistical difference is aobvious
It writes, P < 0.01 item indicates that statistical difference is extremely significant.
3 experimental results
3.1, influence of the FPS to diabetes B mouse weight
As shown in table 2-2, the weight of each group mouse decreased significantly compared with normally group mouse before treating, and difference has
Significant meaning, after treatment 4 weeks, the mouse weight of model group is still declined, and the weight of other several groups of administration group mouse is
Rise.Further, it was observed that model group mouse activity is reduced during drug treatment, perspiration increases, and hair is rough, out
Sweat is more, and other administration groups are then relatively more active, hair smoothing, and state is preferable.
Table 2-2 FPS to diabetes B mouse weight influence (N=10)
Note: N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+LP
+ LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: fermentation rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freezes
Dry powder high dose group；* p < 0.05, * * p < 0.01 are indicated compared with model group；#p < 0.05, ##p < 0.01 are indicated and normal group
It compares.
3.2, influence of the FPS to diabetes B mouse fasting blood-glucose
N.control in Fig. 2: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+
LP+LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-drying
Powder high dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；##p < 0.01 is indicated compared with N.control；
As shown in Fig. 2, before treatment, after the fasting blood-glucose of diabetic mice is significantly higher than normal group, treatment 4 weeks, model group
The fasting blood-glucose of mouse is risen before relatively treating, and the mouse fasting blood-glucose of remaining each treatment group decreased significantly.With mould
Type group is compared, and difference has significant；The blood glucose of rhizoma polygonati fermentation material freeze-dried powder high dose group has extremely aobvious compared to model group difference
Meaning, show rhizoma polygonati fermentation material freeze-dried powder have apparent hypoglycemic effect, and its therapeutic effect have dose-dependant effect
It answers.Compared with rhizoma polygonati group of not fermenting, rhizoma polygonati powder mixture of lactic acid bacteria group, the empty stomach of rhizoma polygonati fermentation material freeze-dried powder under same dose
Blood glucose is lower, but difference is without significant meaning.
3.3, influence of the PS and FPS to diabetes B Mouse oral glucose tolerance
As shown in table 2-3, the oral glucose tolerance of diabetes B mouse is impaired, cannot make quickly after oral glucose
Blood sugar recovery is to base level, and after drug treatment, the impaired glucose tolerance degree of each administration group is alleviated.Wherein rhizoma polygonati ferments
The therapeutic effect of object freeze-dried powder has significant meaning, i.e. area under the glucose curve of rhizoma polygonati fermentation material freeze-dried powder high dose group
(AUC) difference has extremely significant property meaning (p < 0.01) between model group and rhizoma polygonati group of not fermenting.In addition, after glucose load
30min, the blood glucose value of each group mouse have all reached peak, and to when 60min, in addition to model group, remaining is each after glucose load
Treatment group's blood glucose decreased significantly, and rhizoma polygonati fermentation material freeze-dried powder high dose group and model group and rhizoma polygonati group, rhizoma polygonati powder lactic acid
Bacterium mixture group, which is compared, more significant difference (p < 0.05).Illustrate that rhizoma polygonati fermentation material freeze-dried powder alleviates diabetes B grape
The effect of impaired glucose tolerance is substantially better than rhizoma polygonati.
Table 2-3 PS and FPS to diabetes B mouse OGTT, AUC influence (N=10)
Note: N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+LP
+ LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-dried powder
High dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；#p < 0.05, ##p < 0.01 are indicated and N.control phase
Compare；△ p < 0.05, △ △ p < 0.01 is indicated compared with PS；▽ p < 0.05, ▽ ▽ p < 0.01 is indicated compared with FPSH.
3.4, influence of the PS and FPS to diabetes B mouse islets element tolerance
Diabetes B is the diabetes using insulin resistance as Etiological, as shown in Table 2-4, the performance of model group mouse
Apparent Insulin resistance is gone out, that is, after having injected insulin, blood glucose value is and each there is no apparent downward trend
A treatment group mouse blood glucose after insulin injection 30min significantly decreases, blood glucose value bottom out when arriving 120min, yellow
The blood glucose value of essence group gos up most obvious, and its blood glucose value and fermentation rhizoma polygonati low dose group and high dose group have extremely significant difference
(p<0.01)；In addition, the difference of the AUC of rhizoma polygonati fermentation material freeze-dried powder high dose group and rhizoma polygonati group also have significant (p <
0.05).The difference of rhizoma polygonati group and rhizoma polygonati powder mixture of lactic acid bacteria group is without significant meaning.In general, rhizoma polygonati fermentation material freeze-dried powder
Curative effect be better than rhizoma polygonati, and its therapeutic effect difference has significant (p < 0.05).
Table 2-4 PS and FPS to diabetes B mouse ITT, AUC influence (N=10)
Note: N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+LP
+ LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-dried powder
High dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；
#p < 0.05, ##p < 0.01 are indicated compared with N.control；△ p < 0.05, △ △ p < 0.01 is indicated compared with PS
Compared with；▽ p < 0.05, ▽ ▽ p < 0.01 is indicated compared with FPSH.
3.5, influence of the PS and FPS to diabetes B mouse Fasting insulin level and insulin resistance index
In Fig. 3 and Fig. 4, N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati
Group；PS+LP+LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation
Object freeze-dried powder high dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；##p < 0.01 is indicated and N.control phase
Compare；△ p < 0.05, △ △ p < 0.01 is indicated compared with PS；▽ ▽ p < 0.01 is indicated compared with FPSH.
Such as Fig. 3, shown in Fig. 4, the Fasting insulin level of diabetes B model group mouse is increased, insulin resistance index
Also significant to increase, this is related to the insulin resistance of diabetes B.Each treatment group can reduce the empty stomach of diabetes B mouse
Insulin level alleviates insulin resistance, and wherein the effect of rhizoma polygonati fermentation material freeze-dried powder high dose group is best, with rhizoma polygonati group phase
Than rhizoma polygonati fermentation material freeze-dried powder low dose group significant difference (p < 0.05), rhizoma polygonati fermentation material freeze-dried powder high dose group is reducing
Fasting insulin level alleviate insulin resistance in terms of all have extremely significant difference (p < 0.01), rhizoma polygonati fermentation material freeze-dried powder group with
Rhizoma polygonati group, the difference of rhizoma polygonati powder mixture of lactic acid bacteria group also have significant meaning.Illustrate that rhizoma polygonati fermentation material freeze-dried powder is treating 2 types sugar
Curative effect in terms of urine patient's insulin resistance is substantially better than do not ferment rhizoma polygonati and rhizoma polygonati powder mixture of lactic acid bacteria group.
3.6, influence of the PS and FPS to diabetes B mouse glycosylated hemoglobin
N.control in Fig. 5: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+
LP+LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-drying
Powder high dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；##p < 0.01 is indicated compared with N.control；△
△ p < 0.01 is indicated compared with PS；▽ ▽ p < 0.01 is indicated compared with FPSH.
As shown in figure 5, the glycosylated hemoglobin of model group significantly increases, the mouse glycosylated hemoglobin of each treatment group is aobvious
Write decline, no matter the effect of high dose or low dose group is superior to unfermentable rhizoma polygonati group and rhizoma polygonati powder cream for rhizoma polygonati after fermentation
Sour bacterium mixture group, and difference all has extremely significant meaning (p < 0.01).It may be after fermentation, effective component is become
Change, the decline of glycosylated hemoglobin illustrates effect of the rhizoma polygonati after fermentation in terms of control blood glucose for a long time better than the rhizoma polygonati that do not ferment.
3.7, influence of the PS and FPS to diabetes B lipid of mice
As shown in table 2-5, rhizoma polygonati fermentation material freeze-dried powder have reduce the triglycerides of diabetes B mouse, total cholesterol,
The effect of low-density lipoprotein, and effect is better than rhizoma polygonati group of not fermenting.In this experiment, because high cholesterol high in fat has been used to raise
Material, triglycerides, total cholesterol, low-density lipoprotein and the high-density lipoprotein of diabetes B mouse are above normal group,
The indices for the treatment of group have decline, and wherein high-density lipoprotein is also declined slightly but is still above normal group.Show rhizoma polygonati
Fermentation material freeze-dried powder has effect for reducing blood fat.
Table 2-5 PS and FPS to diabetes B lipid of mice influence (N=10)
Note: N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+LP
+ LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-dried powder
High dose group；* p < 0.05, * * p < 0.01 are indicated compared with Model；#p < 0.05, ##p < 0.01 are indicated and N.control phase
Compare.
3.8, influence of the PS and FPS to diabetes B mouse glucagon-like-peptide-1
Glucagon-like-peptide-1 (GLP-1) is a kind of important peptide hormone in body, passes through glucagon suppression
Secretion play blood sugar reducing function, that is, to act on pancreatic islet alpha and β cell can reduce the secretion of glucagon.It can be with from table 2-6
Find out, the secretory volume of the GLP-1 of model group is substantially reduced, and the GLP-1 secretory volume of each treatment group is risen, and wherein rhizoma polygonati is sent out
The therapeutic effect of ferment object freeze-dried powder is best, and compared with rhizoma polygonati group, rhizoma polygonati powder mixture of lactic acid bacteria group, rhizoma polygonati fermentation material freeze-dried powder
High dose group can dramatically increase the secretion (p < 0.05) of GLP-1.
Table 2-6 PS and FPS to diabetes B mice serum glucagon-like peptide 1 content influence (N=
10)
Note: N.control: normal group；Model: model group；P.control: positive controls；PS: rhizoma polygonati group；PS+LP
+ LB: rhizoma polygonati powder mixture of lactic acid bacteria group；FPSL: rhizoma polygonati fermentation material freeze-dried powder low dose group；FPSH: rhizoma polygonati fermentation material freeze-dried powder
High dose group；* p < 0.01 is indicated compared with Model；##p < 0.01 is indicated compared with N.control；△ p < 0.05 is indicated
Compared with PS.
4, conclusion
Based on the above results, it can be seen that rhizoma polygonati and rhizoma polygonati fermentation material freeze-dried powder can significantly improve diabetes B mouse
Oral glucose tolerance is impaired and insulin resistance, can reduce diabetes B mouse fasting blood-glucose and postprandial blood sugar and
Its blood lipid level, while also can be reduced the accumulation of epididymal adipose tissues.Especially from insulin tolerance, Fasting insulin level, saccharification blood
It is small to diabetes B that the testing result of the indexs such as Lactoferrin, glucagon-like peptide 1 can be seen that rhizoma polygonati fermentation material freeze-dried powder
The therapeutic effect of mouse is substantially better than the rhizoma polygonati powder of non-fermentation process, and rhizoma polygonati powder lactic acid bacteria powder mixture and the rhizoma polygonati that do not ferment
No significant difference in index of correlation, response to treatment are suitable.
Involved in claims of the present invention when numberical range, it is thus understood that two endpoints of each numberical range and two
Any one numerical value can be selected between a endpoint, repeat in order to prevent, the present invention describes preferred embodiment, but this field
Technical staff once know basic creative concept, then other change and modification can be made to embodiment.
Obviously, various changes and modifications can be made to the invention by those skilled in the art, without departing from of the invention
Spirit and scope, if in this way, these modifications and changes of the present invention belongs to claim and its equivalent technologies of the invention
Within the scope of, then the present invention is also intended to include these modifications and variations.
Claims (10)
1. a kind of preparation method of rhizoma polygonati fermentation material, which is characterized in that select the lactic acid bacteria class probiotics that can generate aminobutyric acid
As leavening, leavening is seeded in the rhizoma polygonati slurries after sterilizing and is fermented, rhizoma polygonati fermentation material is obtained.
2. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the leavening is by plant cream
Bacillus and Lactobacillus brevis are mixed to prepare according to bacteria suspension volume ratio 1:0-10.
3. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the leavening presses volume hundred
In rhizoma polygonati slurries after dividing the inoculum concentration access sterilizing than 1%-30%, and the 10-80h that ferments at 20-36.5 DEG C.
4. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the pH of the rhizoma polygonati slurries is
6.0 ± 2, when fermentation liquid pH is reduced to 3.0-3.5 to indicate that fermentation is completed.
5. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the preparation of the rhizoma polygonati slurries
Method is as follows: fresh rhizoma polygonati or dry rhizoma polygonati being cleaned, is crushed, is added water, obtain rhizoma polygonati slurries；The sterilized processing of rhizoma polygonati slurries,
Rhizoma polygonati slurries after must sterilizing.
6. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that rhizoma polygonati in the rhizoma polygonati slurries
Dry weight: the mass ratio of water is 1:3~10.
7. a kind of preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the rhizoma polygonati fermentation material is through fast
It after fast prefreezing, is freeze-dried under vacuum conditions, crushes, obtain rhizoma polygonati fermentation material freeze-dried powder.
8. the preparation method of rhizoma polygonati fermentation material as described in claim 1, which is characterized in that the concentration of the bacteria suspension be 1 ×
106-1×108A/mL.
9. the rhizoma polygonati fermentation material that the preparation method of claim 1-8 any one rhizoma polygonati fermentation material is prepared.
10. rhizoma polygonati fermentation material as claimed in claim 9 is in the food, drug or health-oriented products of preparation treatment diabetes B
Application.
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