CN102961733A - Application of seaweed polypeptide in preparing health-care product for reducing blood fat and blood sugar as well as beverage containing polypeptide for reducing blood fat and blood sugar - Google Patents
Application of seaweed polypeptide in preparing health-care product for reducing blood fat and blood sugar as well as beverage containing polypeptide for reducing blood fat and blood sugar Download PDFInfo
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Abstract
The invention relates to an application of a seaweed polypeptide in preparing a health-care product for reducing blood fat and blood sugar as well as a beverage containing the polypeptide for reducing the blood fat and the blood sugar. The seaweed polypeptide is prepared by the following steps of: milling a mixture of immersed porphyra haitanensis and water through a colloid mill to obtain porphyra haitanensis slurry; adopting AS.1398 neutral protease to carry out enzymolysis on the porphyra haitanensis slurry for 5-10 hours under the conditions that the temperature is 45-55 DEG C, the pH (Potential of Hydrogen) is 7.2-7.8, the enzyme adding amount E/S is 8000-12000 U/g and the weight/volume ratio concentration of a substrate is 3-7%, so as to obtain an enzymolysis solution; centrifuging the enzymolysis solution and collecting liquid supernatant; and carrying out ultra-filtering separation on the liquid supernatant to obtain an active component with the molecular weight of less than 10000 Da. An animal experiment shows that agars are subjected to biological enzymolysis to obtain the seaweed polypeptide which has the obvious effects of reducing the blood fat, the blood sugar and blood pressure; and after the seaweed polypeptide is taken for a long period, the weight can be prevented from too fast increase and the happening possibility of arteriosclerosis can be reduced.
Description
Technical field
The present invention relates to the medical usage and the beverage that comprises aforementioned polypeptides of polypeptide, relate to particularly a kind of blood fat reducing, blood sugar lowering beverage that has the Sargassum polypeptide of blood fat reducing, function of blood sugar reduction and comprise aforementioned polypeptides.
Background technology
Sargassum belongs to rudimentary plant, mainly is grown in neritic province domain, China's seashore line length, and the shallow sea area is large, the Sargassum abundant species, this wherein has the Sargassum of economic worth mainly to be divided into chlorella, Brown algae and red algae three major types.At present, the economical alga of having carried out artificial cultivation mainly contains: the Thallus Laminariae (Thallus Eckloniae), Thallus Laminariae and the Sargassum fusiforme (Harv.) Setch that belong to Brown algae; Porphyra haitanensis, Porphyra yezoensis Ueda, Thallus Gracilariae, Gracilaria tenuistipitata, Bangia fuscopurpurea and the Eucheuma muricatum (Gmel.) Web.Van Bos. etc. that belong to red algae.
Yet because China's Sargassum career development is in the junior stage, therefore, its utilization for Sargassum is quite inadequate, and at present, the seaweed products of China concentrate on this one side of Sargassum polysaccharides.According to bibliographical information, the Sargassum polysaccharides in the Sargassum all has and improves body immunity, anticancer, antiviral activity, and can reduce and cause atherosclerotic lipid content in the blood vessel.Therefore, the industrialization seaweed products of domestic market comparative maturity are mainly the Sargassum polysaccharides product: the agar that extracts from red algae, carrageenan etc., the Algin that extracts from Brown algae, fucoidan etc., and the xylan that extracts from chlorella, mannan etc.And Sargassum polysaccharides is the biomacromolecule that a class forms very complex, its normal ethanol precipitation, the quaternary amine sedimentation method, DEAE fibre column chromatography and gel filtration chromatography method of adopting of purifying, and can reach the purpose of polysaccharide and Protein Separation by molecular sieve or anion-exchange column etc., and then so that the Sargassum protein that accounts between the Sargassum content 10%~50% has formed industrial waste mostly, cause significant wastage.
On the other hand, existing hypoglycemic medicine has insulin, biguanides (representing medicine is metformin), sulphanylureas (representing medicine is glipizide, gliclazide etc.), alpha-glucosidase inhibitor (representing medicine is acarbose), euglycemic agent (representing medicine is rosiglitazone, pioglitazone etc.).Yet above-mentioned various medicines have its limitation, and are larger to the injury of the intestines and stomach such as the antidiabetic drug of biguanides, cause easily dyspepsia; Sulfonylurea drugs may cause leukopenia; Alpha-glucosidase inhibitor then can produce gastrointestinal reaction, and concrete manifestation has abdominal distention, flatulence, epigastrium causalgia, diarrhoea or constipation etc.; Euglycemic agent then might cause the generation of water retention.
And existing the most frequently used blood lipid-lowering medicine is HMG-CoA reductase inhibitor, lovastatin, simvastatin, pravastatin, mevastatin, fluvastatin, atorvastatin, cerivastatin and rosuvastatin etc. are specifically arranged, yet, statins causes that easily the liver enzyme increases, may cause changes of liver function, and some patients were has the symptom of myalgia after taking medicine, indivedual even initiation sarcolysis disease.
To sum up, there is no a kind of reliable, gratifying Therapeutic Method for blood sugar lowering, blood fat reducing at present.
Summary of the invention
The object of the present invention is to provide the purposes of Sargassum polypeptide in preparation blood fat reducing, hypoglycemic drug.
The purposes of Sargassum polypeptide in preparation blood fat reducing, health-caring product capable of reducing blood sugar, described Sargassum polypeptide adopts following mode to make: the porphyra haitanensis after the immersion and the mixed material of water grind through colloid mill, obtain the porphyra haitanensis serosity, adopt the AS.1398 neutral protease at 45 ℃~55 ℃, pH7.2~7.8, enzyme concentration E/S 8000~12000U.g
-1, concentration of substrate 3~7%(w/v) condition under enzymolysis porphyra haitanensis serosity 5h~10h, obtain enzymolysis solution, with the centrifugal rear collection supernatant of enzymolysis solution, again supernatant is obtained molecular weight less than the active component of 10000 Da, freeze-dried back through ultra-filtration and separation.
Porphyra haitanensis of the present invention (
Porphyra haitanensis) belong to the former Rhodophyceae of Rhodophyta (Rhodophyta) (Protoflorideae) Bangiales (Bangiales) Bangia fuscopurpurea section (Bangiaceae) Porphyra (Porphyra).
In the embodiment that recommends, preferably adopt the AS.1398 neutral protease at 50 ℃, pH7.5, enzyme concentration E/S 10000U.g
-1, concentration of substrate 5%(w/v) condition under enzymolysis porphyra haitanensis serosity 10h, obtain enzymolysis solution.
In the embodiment that recommends, the supernatant of centrifugal rear collection is by one times of the ratio thin up of 1:1, and the supernatant adding ultrafiltration apparatus after then will diluting separates.
Another object of the present invention is to provide the blood fat reducing, the blood sugar lowering beverage that comprise above-mentioned Sargassum polypeptide.
A kind of blood fat reducing, blood sugar lowering beverage that comprises the Sargassum polypeptide, it is characterized in that: it comprises Sargassum polypeptide and the edible drink formula of blood fat reducing, blood sugar lowering effective dose, described Sargassum polypeptide stock solution adopts following mode to make: the porphyra haitanensis after the immersion and the mixed material of water grind through colloid mill, obtain the porphyra haitanensis serosity, adopt the AS.1398 neutral protease at 45 ℃~55 ℃, pH7.2~7.8, enzyme concentration E/S 8000~12000U.g
-1, concentration of substrate 3~7%(w/v) condition under enzymolysis porphyra haitanensis serosity 5h~10h, obtain enzymolysis solution, with the centrifugal rear collection supernatant of enzymolysis solution, again supernatant is obtained molecular weight less than the Sargassum polypeptide stock solution of the active component of 10000 Da through ultra-filtration and separation.
The present invention removes Sargassum polypeptide liquid bitter off-flavors by above-mentioned separation method, and keeps strong Thallus Porphyrae local flavor, and then obtains the beverage products that mouthfeel is unique, have food therapy value.
In the embodiment that recommends, described drink formula comprises cyclodextrin, malic acid, sodium citrate, sodium isoascorbate, Vc, glycine, white sugar and sodium carboxymethyl cellulose, and adds in the Sargassum polypeptide stock solution according to the ratio of cyclodextrin 5 ‰~7 ‰, malic acid 0.5 ‰~1 ‰, sodium citrate 0.2 ‰~0.6 ‰, sodium isoascorbate 0.2 ‰~0.6 ‰, Vc 0.2 ‰~0.6 ‰, glycine 0.1 ‰~0.3 ‰, white sugar 50 ‰~80 ‰ and sodium carboxymethyl cellulose 0.5~0.8 ‰.
In the embodiment that recommends, described drink formula also comprises maple sugar essence and roasting barley essence, and adds in the Sargassum polypeptide stock solution according to the ratio of maple sugar essence 0.1 ‰~0.3 ‰ and roasting barley essence 0.5 ‰~1.5 ‰.
Need to prove that raw material used in the present invention is dried porphyra haitanensis, it both can select the first planting Thallus Porphyrae that mouthfeel is better, nutritive value is higher, also can use the lower end stubble Thallus Porphyrae of mouthfeel and nutritive value.And can be take the Thallus Porphyrae of four batches or five batches as raw material in view of the Sargassum polypeptide beverage, therefore, its raw material sources are wide, cost is low, preparation process is simple, equipment and fund input are relatively less, and the significant added value of improving product, obtain higher economic benefit.
Sargassum polypeptide beverage provided by the invention has following features:
1, taste lubrication, sour-sweet moderate, Thallus Porphyrae is with rich flavor, is royal purple, transparent clear.It is suitable for various fill, can reach more than 1 year through the shelf-life behind the ultra high temperature short time sterilization;
2, organic iodine rich content is suitable for the utilization and extention of the landlocked iodine deficient area of China, plays the effect of the abundant product that enriches the iodine; Vitamin and content of mineral substances are also higher, and fishy smell is little, unique flavor, are the healthy marine foods of typical low fat, rich cellulose and mineral;
3, the effect that has blood fat reducing, blood sugar lowering and blood pressure lowering: confirm by zoopery, Thallus Porphyrae has significant blood fat reducing, blood sugar lowering and hypotensive activity through the Sargassum polypeptide that biological enzymolysis obtains, long-term taking can suppress the body weight excessively rapid growth, (report about the hypotensive activity of Sargassum polypeptide is asked for an interview the Chinese Sea medicine to reduce arteriosclerotic Probability, 03 phase in 2010, Wang Yin etc.);
The present invention utilizes the marine algae resource of China's abundant, theoretical and modern scientific research achievement is basic take Chinese traditional methods to keep in good health, take the dietetic therapy homology as theory, according to the trophic function characteristic of Sargassum and the demand of modern diet structure, has the activity polypeptid substance of blood fat reducing, effect of lowering blood sugar by extractions such as modern biotechnology, nanometer film hyperfiltration technique, Analysis on Biological Activity technology.The Sargassum polypeptide series products of the present invention's development can be filled up the market vacancy of Sargassum protein active polypeptide product, and because the Sargassum active polypeptide self has the advantages such as easy absorption, high activity, blood fat reducing, blood sugar lowering, blood pressure reduction effect be remarkable, distinctive algae fragrance is fit to present people demand to health, nutrition, delicious food aspect food in addition.
The present invention is take this Sargassum polypeptide as main prescription, inquire into the technology that removes of Sargassum polypeptide liquid bitter off-flavors, the allocating technology of local flavor, optimize condition and the best sterilization scheme affect product stability, be refined into the Novel series seaweeds such as the Sargassum polypeptide beverage health goods of health-care effect dietetic therapy homologies such as having nutrition and blood fat reducing, blood sugar lowering, blood pressure lowering concurrently.This meets the direction of current alga food research and development, meet Marine health care food potential market demand, has huge promotion prospect, can promote the Sargassum industry to go on the road of the sound development of friendly process, green product, drive Sargassum industrial economy benefit and increase, promote the development of China's marine economy.
The specific embodiment
For the ease of understanding the present invention, the spy enumerates embodiment, with further annotation the present invention, rather than to the restriction of any mode of the present invention.
Embodiment 1
1) raw material: the dried porphyra haitanensis of the material choice first planting;
2) soak: in the 1:25(porphyra haitanensis: ratio water) adds water, soaks 30 min;
3) colloid mill rubs: porphyra haitanensis is soaked mixed liquor by colloid mill, form the thinner porphyra haitanensis serosity of granule;
4) heat tracing: the porphyra haitanensis serosity is heated to central temperature reaches 50 ℃;
5) protease hydrolyzed: adopt the AS.1398 neutral protease at 50 ℃, pH7.5, enzyme concentration E/S 10000U.g
-1, concentration of substrate 5%(w/v) condition under enzymolysis porphyra haitanensis serosity 10h, obtain enzymolysis solution;
6) centrifugal: with enzymolysis solution in 8000r.min
-1Centrifugal 10min tentatively removes the Thallus Porphyrae slag, obtains supernatant;
7) supernatant dilution: the centrifugal supernatant of collecting is comparatively dense thick, first in one times of the ratio thin up of 1:1;
8) ultrafiltration remove impurity: the supernatant after will diluting separates by the ultrafiltration apparatus of the pottery volume heart, obtains molecular weight less than the active component of 10000 Da, freeze-dried back.
Embodiment 2
1) raw material: the 3rd batch of dried porphyra haitanensis of material choice;
2) soak: in the 1:25(porphyra haitanensis: ratio water) adds water, soaks 30 min;
3) colloid mill rubs: porphyra haitanensis is soaked mixed liquor by colloid mill, form the thinner porphyra haitanensis serosity of granule;
4) heat tracing: the porphyra haitanensis serosity is heated to central temperature reaches 45 ℃;
5) protease hydrolyzed: adopt the AS.1398 neutral protease at 45 ℃, pH7.2, enzyme concentration E/S 8000U.g
-1, concentration of substrate 3%(w/v) condition under enzymolysis porphyra haitanensis serosity 10h, obtain enzymolysis solution;
6) centrifugal: with enzymolysis solution in 8000r.min
-1Centrifugal 10min tentatively removes the Thallus Porphyrae slag, obtains supernatant;
7) supernatant dilution: the centrifugal supernatant of collecting is comparatively dense thick, first in one times of the ratio thin up of 1:1;
8) ultrafiltration remove impurity: the supernatant after will diluting separates by the ultrafiltration apparatus of the pottery volume heart, obtains molecular weight less than the active component of 10000 Da, and is for subsequent use after the spray drying.
Embodiment 3
1) raw material: the dried porphyra haitanensis of material choice end stubble;
2) soak: in the 1:30(porphyra haitanensis: ratio water) adds water, soaks 30 min;
3) colloid mill rubs: porphyra haitanensis is soaked mixed liquor by colloid mill, form the thinner porphyra haitanensis serosity of granule;
4) heat tracing: the porphyra haitanensis serosity is heated to central temperature reaches 55 ℃;
5) protease hydrolyzed: adopt the AS.1398 neutral protease at 55 ℃, pH7.8, enzyme concentration E/S 12000U.g
-1, concentration of substrate 7%(w/v) condition under enzymolysis porphyra haitanensis serosity 5h, obtain enzymolysis solution;
6) centrifugal: with enzymolysis solution in 8000r.min
-1Centrifugal 10min tentatively removes the Thallus Porphyrae slag, obtains supernatant;
7) supernatant dilution: the centrifugal supernatant of collecting is comparatively dense thick, first in one times of the ratio thin up of 1:1;
8) ultrafiltration remove impurity: the supernatant after will diluting separates by the ultrafiltration apparatus of the pottery volume heart, obtains molecular weight less than the active component of 10000 Da, freeze-dried back.
Embodiment 4
1) raw material: the 3rd batch of dried porphyra haitanensis of material choice;
2) soak: in the 1:20(porphyra haitanensis: ratio water) adds water, soaks 30 min;
3) colloid mill rubs: porphyra haitanensis is soaked mixed liquor by colloid mill, form the thinner porphyra haitanensis serosity of granule;
4) heat tracing: the porphyra haitanensis serosity is heated to central temperature reaches 45 ℃;
5) protease hydrolyzed: adopt the AS.1398 neutral protease at 45 ℃, pH7.2, enzyme concentration E/S 8000U.g
-1, concentration of substrate 3%(w/v) condition under enzymolysis porphyra haitanensis serosity 10h, obtain enzymolysis solution;
6) centrifugal: with enzymolysis solution in 8000r.min
-1Centrifugal 10min tentatively removes the Thallus Porphyrae slag, obtains supernatant;
7) supernatant dilution: the centrifugal supernatant of collecting is comparatively dense thick, first in one times of the ratio thin up of 1:1;
8) ultrafiltration remove impurity: the ultrafiltration apparatus that the supernatant after will diluting passes through the pottery volume heart separates, and obtains molecular weight less than the Sargassum polypeptide stock solution of the active component composition of 10000 Da;
9) ratio according to cyclodextrin 6.5 ‰, malic acid 0.8 ‰, sodium citrate 0.4 ‰, sodium isoascorbate 0.45 ‰, Vc 0.5 ‰, glycine 0.15 ‰, white sugar 70 ‰ and sodium carboxymethyl cellulose 0.6 ‰ adds above-mentioned substance in Sargassum polypeptide stock solution, dissolving obtains the Sargassum polypeptide beverage;
10) sterilization is canned: canned material is selected the tinplate built-up tin, and pasteurize (85 ℃, 20min) obtains beverage products.
The beverage taste is tried out and ocular estimate is finished by the assessment panel that several professionals form, this beverage mouthfeel lubricated, sour-sweet moderate, Thallus Porphyrae is with rich flavor, is not with bitter off-flavors, outward appearance to be royal purple, transparent clear.
Embodiment 5
1) raw material: the dried porphyra haitanensis of material choice end stubble;
2) soak: in the 1:25(porphyra haitanensis: ratio water) adds water, soaks 30 min;
3) colloid mill rubs: porphyra haitanensis is soaked mixed liquor by colloid mill, form the thinner porphyra haitanensis serosity of granule;
4) heat tracing: the porphyra haitanensis serosity is heated to central temperature reaches 55 ℃;
5) protease hydrolyzed: adopt the AS.1398 neutral protease at 55 ℃, pH7.8, enzyme concentration E/S 12000U.g
-1, concentration of substrate 7%(w/v) condition under enzymolysis porphyra haitanensis serosity 5h, obtain enzymolysis solution;
6) centrifugal: with enzymolysis solution in 8000r.min
-1Centrifugal 10min tentatively removes the Thallus Porphyrae slag, obtains supernatant;
7) supernatant dilution: the centrifugal supernatant of collecting is comparatively dense thick, first in one times of the ratio thin up of 1:1;
8) ultrafiltration remove impurity: the ultrafiltration apparatus that the supernatant after will diluting passes through the pottery volume heart separates, and obtains molecular weight less than the Sargassum polypeptide stock solution of the active component composition of 10000 Da;
9) ratio according to cyclodextrin 6.5 ‰, malic acid 0.8 ‰, sodium citrate 0.4 ‰, sodium isoascorbate 0.45 ‰, Vc 0.5 ‰, glycine 0.15 ‰, white sugar 70 ‰, sodium carboxymethyl cellulose 0.6 ‰, maple sugar essence 0.1 ‰ and roasting barley essence 1 ‰ adds above-mentioned substance in Sargassum polypeptide stock solution, dissolving obtains the Sargassum polypeptide beverage; This wherein, maple sugar essence, roasting barley essence are the perfumery prescription, therefore, can allocate other odor type according to consumer taste;
10) sterilization is canned: canned material is selected the tinplate built-up tin, and pasteurize (85 ℃, 20min) obtains beverage products.
The beverage taste is tried out and ocular estimate is finished by the assessment panel that several professionals form, this beverage mouthfeel lubricated, sour-sweet moderate, Thallus Porphyrae is with rich flavor, is not with bitter off-flavors, outward appearance to be royal purple, transparent clear.
Test example 1
1 experiment material and method
1.1 Sargassum polypeptide:
The Sargassum polypeptide that embodiment 1 makes.
1.2 laboratory animal:
Kunming mice (KM), male, weight 20~25g, 100;
The Wistar rat, male, weight 150~200g, 60.
1.3 index detectable:
T-CHOL test kit, Triglyceride Reagent box, HDL-C test kit, low-density lipoprotein cholesterol test kit, superoxide dismutase test kit, glutathion peroxidase test kit, Malondialdehyde Kit, nitric oxide test kit; Glycated serum protein test kit, serum insulin test kit, liver glycogen test kit etc.
1.4 other reagent materials:
Simvastatin, the Beijing WBL Peking University Biotech Co., Ltd; Glipizide controlled release tablets, Pfizer pharmaceutical Co. Ltd; Alloxan, sigma company; High lipid food (prescription: 10% Adeps Sus domestica, 10% yolk powder, 1% cholesterol, 0.3% cholate, 78.7% normal feedstuff), the two lion laboratory animal feed technology company limiteies in Suzhou.
1.5 effect for reducing blood fat animal experiment method:
1.5.1 laboratory animal grouping
After 60 Wistar rats are fed 5d with the normal feedstuff adaptability, claim weight and carry out the stratified random grouping according to weight, comprise 5 groups of hyperlipidemia model Wistar rats (50,10 every group) and 1 group of Normal group (10).
1.5.2 hyperlipidemia model rat modeling
5 groups of hyperlipidemia model Wistar rats are carried out hyperlipidemia model modeling test, and 50 Wistar rats are fed 2 weeks of high lipid food continuously, the Normal group normal feedstuff of feeding, and afterbodys are got blood after 2 weeks, and the index that detects serum TC, TG, HDL-C, LDL-C detects.
1.5.3 experiment grouping and dosage design
Choose 5 groups in the hyperlipidemia model Wistar rat, group is respectively B hyperlipidemia model matched group, the high fat medicine of C matched group (simvastatin 10mgkg
-1), the high fat Sargassum of D polypeptide low dose group (0.5 gkg
-1), dosage group (1.0 gkg among the E
-1), F high dose group (1.5 gkg
-1); The group of Normal group is the A Normal group.Concrete grouping and dosage require as shown in table 1.
The grouping of table 1 lipid-lowering test animal and dosage
Sequence number | Group | Feed for nursing | The gavage medicine | Dosage (mg/kg bw) |
A | Normal group | Normal feedstuff | Distilled water | - |
B | The hyperlipidemia model matched group | High lipid food | Distilled water | - |
C | High fat medicine matched group | High lipid food | Simvastatin | 10 |
D1 | Sargassum polypeptide low dose group | High lipid food | The Sargassum polypeptide | 500 |
D2 | Dosage group in the Sargassum polypeptide | High lipid food | The Sargassum polypeptide | 1000 |
D3 | Sargassum polypeptide high dose group | High lipid food | The Sargassum polypeptide | 1500 |
1.5.4 feeding manner
Every group of 10 Wistar rat are that 1 mouse cage is raised, and 6 groups of experiments are carried out simultaneously, according to the design of the table 1 different feedstuff of feeding, normally feed water.
1.5.5 administering mode
Administration every day before measurement rat weight according to the different dosing dosage of different groups, is dissolved in the distilled water of equal volume, with the mode administration of gavage.Timing every day gastric infusion is (about at 10 in the morning) once, and wherein, A group and B organize these two groups of matched groups equal volume distilled water gavage.Test continuous 4 weeks, the experimental session rat freely drinks water and ingests.
1.5.6 DATA REASONING
Experimental data mainly comprises the liver coefficient of serum lipids index (TC, TG, HDL-C, LDL-C), liver organization of rat after respectively organizing rat weight data, experiment in 30 days finishes and Antioxidant Indexes (SOD, MDA) etc.
Weight is measured: measure rat weight before the gastric infusion every day, and that treating excess syndrome is tested is front, the weight data in 1 week, 2 weeks, 3 weeks, 4 weeks are added up the changes of body mass of analyzing rat.
Serum Indexes is measured: fasting evening before that day of last gavage, pluck eye behind gavage 1 h and get blood (every rat is got 3mL blood), blood sample extracts serum (every rat 0.5mL) in the room temperature hold over night, adopts TC, TG, the indexs such as HDL-C, LDL-C in the kit measurement serum.
The liver index measurement: after experiment finished, animal was put to death in the vertebra dislocation, went liver organization 2g with the remaining bloodstain of washing immediately, fully ground in 4 ℃ of ice baths, and liver organization is in 8000 rmin
-1, 4 ℃ of centrifugal 10min, with the indexs such as SOD, MDA in the kit measurement liver organization.
1.6 hypoglycemic activity animal experiment method:
1.6.1 the diabetes mice model is set up
The KM Mus normal diet of feeding, adapt to 3d after, fasting 12h, after weighing, lumbar injection 200mg/kg bw alloxan.Behind the 3d, fasting 5h, tail point blood sampling is surveyed fasting blood sugar with blood glucose meter, blood glucose value〉16mmol/L be considered as the modeling success.
1.6.2 grouping
Survey blood glucose value after the modeling, the mice of screening modeling success.Diabetic mice is divided into 5 groups at random, 10 every group, is respectively: dosage group, E3 Sargassum polypeptide high dose group in B hyperglycemia model group, C medicine matched group, E1 Sargassum polypeptide low dose group, the E2 Sargassum polypeptide; In addition 10 of random chooses not the mice of modeling be the A Normal group.Concrete grouping and dosage such as following table 2:
The grouping of table 2 blood sugar lowering laboratory animal and dosage
Sequence number | Group | Medicine | Dosage (mg/kg bw) |
A | Normal group | Distilled water | —— |
B | The hyperglycemia model group | Distilled water | —— |
C | The medicine matched group | Glipizide controlled release tablets | 10 |
E1 | Sargassum polypeptide low dose group | The Sargassum polypeptide | 500 |
E2 | Dosage group in the Sargassum polypeptide | The Sargassum polypeptide | 1000 |
E3 | Sargassum polypeptide high dose group | The Sargassum polypeptide | 1500 |
1.6.3 administration is raised
Administration every day before measurement rat weight according to the grouping dosage of table 2, is dissolved in the distilled water of equal volume, with the mode administration of gavage.Regularly be administered once every day, and wherein, A group and B organize these two groups of matched groups equal volume distilled water gavage.Normal forage feed, normal feedwater continued for 4 weeks.
1.6.4 DATA REASONING
Experimental data mainly comprises respectively organizes weight, fasting blood sugar, load carbohydrate tolerance, organ index etc.
Weight: measure KM Mus weight every day before the gastric infusion, analyzes the changes of body mass of KM Mus.
The fasting glucose pH-value determination pH: behind the KM Mus fasting 5h, the blood sampling of tail point is surveyed fasting blood sugar with blood glucose meter.
Load carbohydrate tolerance test: test last day, the fasting 5h of KM Mus elder generation, again the medicament of gavage corresponding dosage, behind the 20min, gavage 2.0g/kg bw glucose, afterwards in 0h, 0.5h, 2h, respectively afterbody blood sampling, measure blood glucose value, observe each group to the variation of area under each time point blood glucose line behind the glucose.Area under the curve of blood glucose=0.25 * (0h blood glucose value+4 * 0.5h blood glucose value+3 * 2h blood glucose value).
Organ index is measured: after experiment finished, mice was weighed, get blood after, open abdomen and get Thymus and spleen, use scales/electronic balance weighing.Organ index %=(internal organs wet quality g ÷ weight g) * 100%
2 experimental results and analysis
2.1 the effect for reducing blood fat of Sargassum polypeptide research
2.1.1 the Sargassum polypeptide is on the impact of rat weight
Raising and experiment through continuous 4 weeks, the weight of Wistar rat all increases to some extent, wherein the Weight gain amplitude of hyperlipidemia model group rat is maximum, and high lipid food is fed, and weights reach (313.90 ± 12.91) g after 4 weeks, exceeds 8.5% than the rats in normal control group of the normal feedstuff of feeding.Each dosage group of medicine matched group and Sargassum polypeptide is compared with the hyperlipidemia model group, and its weight is to some extent decline (table 3) also.Data show that the Sargassum polypeptide is the increase of control volume quality effectively.
Table 3 Sargassum polypeptide is on the impact (g) of hyperlipidemia model rat weight
Before the experiment | 1 week | 2 weeks | 3 weeks | 4 weeks | |
The A Normal group | 192.66±7.30 | 231.50±11.04 | 252.50±7.56 | 269.40±8.76 | 288.50±7.55 |
B hyperlipidemia model group | 195.58±8.72 | 241.60±12.87 | 272.40±14.77 | 293.40±14.80 | 313.90±12.91 |
C medicine matched group | 195.78±8.57 | 239.78±16.77 | 260.56±18.95 | 280.67±18.41 | 289.44±14.10 |
E1 Sargassum polypeptide low dose group | 193.40±9.23 | 230.50±16.94 | 258.80±16.58 | 272.90±18.92 | 287.90±19.81 |
Dosage group in the E2 Sargassum polypeptide | 193.02±9.2 | 230.50±17.33 | 250.00±17.64 | 267.90±18.76 | 284.60±17.41 |
E3 Sargassum polypeptide high dose group | 192.36±9.07 | 236.50±13.98 | 261.80±17.31 | 280.70±14.80 | 293.10±14.04 |
2.1.2 the Sargassum polypeptide is on the impact of rat blood serum blood lipid level
The indexs such as the cholesterol in the serum, triglyceride, low-density lipoprotein cholesterol and HDL-C are to weigh the significant data of Blood Lipid.From the data of table 4, can find out, experiment through 4 weeks, the content of the cholesterol of hyperlipidemia model group, triglyceride, this 3 index of low-density lipoprotein cholesterol all is higher than Normal group, HDL-C content is lower than Normal group, the hyperlipidemia model modeling success of Wistar rat is described.Each dosage group of medicine matched group and Sargassum polypeptide with the hyperlipidemia model group relatively also can be found out index content such as having cholesterol reducing, triglyceride, low-density lipoprotein cholesterol and the effect that improves HDL-C content.Simultaneously, find that the regulating effect of Sargassum polypeptide high dose group is best, have certain dosage effect between each dosage group.
Table 4 Sargassum polypeptide is on the impact (mmol/L) of hyperlipidemia model rat blood serum blood lipid level
2.1.3 the Sargassum polypeptide is on the impact of rat liver index and lipid peroxidation index of correlation thereof
The rat high lipid food was fed after 4 weeks, and liver indexes of comparing other each groups with Normal group all have significant increase (table 5).Simultaneously, find in opening the abdominal cavity anatomic observation that the normal liver scarlet with rats in normal control group, smooth, that texture is fine and smooth is compared, other the obvious enlargement of liver, color dimness, textures of respectively organizing rat are rough, surperficial with fat deposit.As seen.The rat hyperlipidemia that high lipid food causes is comparatively serious to the infringement of liver.Lipid peroxidation index data show from liver, compare with Normal group, the malonaldehyde of hyperlipidemia model group (MDA) content exceeds 1.2 times, reach (14.81 ± 1.41) nmol/mgprot, the MDA content of each dosage group of Sargassum polypeptide decreases, and wherein high dose group is compared reduction amplitude maximum with the hyperlipidemia model group.And compare with Normal group, the superoxide dismutase of hyperlipidemia model group (SOD) content significantly descends, and the SOD content of each dosage group of Sargassum polypeptide is higher than the hyperlipidemia model group.As seen, Long-term Feeding Sargassum polypeptide, the MDA content in the functions for the treatment of hyperlipidemia rat liver, and increased SOD content.
Table 5 Sargassum polypeptide is on the impact of rat liver index and lipid peroxidation index of correlation thereof
? | Liver index (%) | MDA(nmol/mgprot) | SOD(U/mgprot) |
The A Normal group | 2.92±0.22 | 6.67±1.41 | 63.65±5.3 |
B hyperlipidemia model group | 3.65±0.21 | 14.81±1.41 | 45.89±5.16 |
C medicine matched group | 3.58±0.17 | 12.55±0.99 | 53.48±2.87 |
E1 Sargassum polypeptide low dose group | 3.47±0.40 | 14.35±1.60 | 49.83±5.19 |
Dosage group in the E2 Sargassum polypeptide | 3.86±0.45 | 13.38±0.97 | 48.19±5.06 |
E3 Sargassum polypeptide high dose group | 3.62±0.27 | 12.62±0.27 | 49.33±3.78 |
2.2 the hypoglycemic activity of Sargassum polypeptide research
2.2.1 the foundation of mice hyperglycemia model
Before test, give the injected in mice alloxan and set up hyperglycemia model, the mouse blood sugar value of modeling reaches (21.23 ± 5.9) mmol/L, and the blood glucose value of Normal group mice is (6.73 ± 1.16) mmol/L, and data show the modeling success.In experimentation, find that hyperglycemia model group mice has obvious polyuria, polydipsia, polyphagia, build to become thin, hair tarnishes, and the curling oneself up hogback does not have the symptoms such as spirit; And the Normal group mental status is good, and bright and clean by hair, growth promoter is normal.
2.2.2 the Sargassum polypeptide is on the impact of Mice Body quality and organ index
After the modeling success, continue the laboratory observation in 4 weeks, find that the weight of hyperlipidemia model group, medicine matched group and each dosage group of Sargassum polypeptide all increasess slowly by (table 6), 4 body mass ratio Normal groups when all are low respectively 28.3%~35.3%.
Table 6 Sargassum polypeptide is on the impact (g) of Mice Body quality
? | After the modeling | 1 week | 2 weeks | 3 weeks | 4 weeks |
The A Normal group | 36.89±1.87 | 40.85±2.91 | 43.74±3.14 | 45.58±3.12 | 46.41±2.74 |
B hyperglycemia model group | 24.72±1.96 | 28.11±2.87 | 29.64±3.45 | 31.09±3.51 | 30.03±3.73 |
C medicine matched group | 25.31±1.79 | 29.10±2.41 | 31.60±2.91 | 32.82±3.68 | 33.28±4.04 |
E1 Sargassum polypeptide low dose group | 24.44±1.79 | 26.98±2.41 | 29.67±2.91 | 31.85±3.68 | 31.11±4.04 |
Dosage group in the E2 Sargassum polypeptide | 24.26±2.59 | 26.70±3.58 | 28.22±4.15 | 29.20±4.24 | 30.61±5.12 |
E3 Sargassum polypeptide high dose group | 24.68±2.74 | 29.45±2.90 | 29.67±2.32 | 30.71±4.12 | 30.21±3.07 |
Diabetes are a kind of chronic dysbolismus diseases, and lasting hyperglycemia can cause immune system disorder, and spleen and the thymus important immune organ that is human body.As shown in Table 7, the spleen of hyperglycemia model group and thymus coefficient all are lower than Normal group, and the obvious atrophy of spleen and thymus is described, immunologic function may descend.Spleen and the thymus coefficient of the middle and high dosage group of medicine matched group and Sargassum polypeptide all are higher than the hyperglycemia model group, illustrate that the Sargassum polypeptide of doses has certain protective effect to spleen and thymus.Liver is one of glycometabolic major organs, it can promote the synthetic of glycogen and store, by finding out in table 7 data, the liver coefficient of hyperglycemia model group mice is apparently higher than Normal group, and the liver coefficient of medicine matched group and each dosage group of Sargassum polypeptide all is lower than the hyperglycemia model group.
The organ coefficient of table 7 model mice (%)
The liver coefficient | Spleen coefficient | The thymus coefficient | |
The A Normal group | 4.20±0.28 | 0.36±0.07 | 0.20±0.04 |
B hyperglycemia model group | 5.26±0.29 | 0.24±0.08 | 0.13±0.05 |
C medicine matched group | 4.92±0.52 | 0.38±0.05 | 0.15±0.05 |
E1 Sargassum polypeptide low dose group | 4.78±0.51 | 0.24±0.07 | 0.12±0.07 |
Dosage group in the E2 Sargassum polypeptide | 5.03±0.64 | 0.31±0.07 | 0.17±0.07 |
E3 Sargassum polypeptide high dose group | 5.03±0.16 | 0.32±0.06 | 0.16±0.06 |
2.2.3 the Sargassum polypeptide is on the impact of mouse's blood sugar content
Through the lasting experiment in 4 weeks, the blood glucose value of mice is carried out tracing observation, the result is as shown in table 8.Data from table can find out that along with the passing of experimental period, the blood glucose value of hyperglycemia model group continues to increase, and blood glucose value reaches (30.29 ± 3.77) mmol/L during 4 week; The blood glucose value continuous decrease of medicine matched group, (21.24 ± 6.08) mmol/L during from the experiment beginning reduces to (15.37 ± 4.61) mmol/L after 4 weeks; The blood glucose value of each dosage group of Sargassum polypeptide is compared hyperglycemia model group blood glucose value and is also all decreased, and wherein the reduction amplitude of the blood glucose value of high dose group is maximum, reduces to (20.59 ± 2.63) mmol/L after 4 weeks, reduces by 32.02% than the hyperglycemia model group.As seen, the Sargassum polypeptide has certain inhibitory action to the blood sugar increasing of hyperglycemia model mice.
The variation (mmol/L) of table 8 hyperglycemia model mouse blood sugar value
After the modeling | 1 week | 2 weeks | 3 weeks | 4 weeks | |
The A Normal group | 6.73±1.16 | 8.04±1.53 | 7.26±1.11 | 8.04±1.50 | 7.61±1.10 |
B hyperglycemia model group | 21.79±6.87 | 24.27±4.23 | 28.65±1.38 | 29.78±1.87 | 30.29±3.77 |
C medicine matched group | 21.24±6.08 | 19.70±5.40 | 18.65±6.55 | 18.51±4.64 | 15.37±4.61 |
E1 Sargassum polypeptide low dose group | 21.03±5.87 | 24.22±5.56 | 26.52±3.91 | 27.40±3.71 | 26.73±3.72 |
Dosage group in the E2 Sargassum polypeptide | 21.28±6.85 | 22.2±5.23 | 23.65±4.48 | 25.09±4.61 | 24.01±3.61 |
E3 Sargassum polypeptide high dose group | 20.82±5.42 | 24.66±2.75 | 23.14±3.20 | 19.91±1.64 | 20.59±2.63 |
2.2.4 the Sargassum polypeptide is on the impact of glucose tolerance in mice
By table 9 data as can be known the Normal group mice after the gavage glucose in the 0.5h blood glucose value reach maximum, return to base level behind the 2h, show as NGT.Hyperglycemia model group mice 0.5h blood glucose value after gavage reaches the highest, but blood glucose value has only fallen 7.1% after rise behind the 2h.The blood glucose value of medicine matched group has fallen 31.2% after rise behind 2h.What falling was maximum behind each dosage group 2h of Sargassum polypeptide is high dose group, has fallen 26.9% after rise.Simultaneously, the Area under the curve of blood glucose of each dosage group of Sargassum polypeptide all is lower than the hyperglycemia model group, and wherein high dose group is minimum, and is close with the medicine matched group, and visible Sargassum polypeptide is the blood sugar lowering area under curve significantly.
Table 9 Sargassum polypeptide is on the impact (mmol/L) of glucose tolerance in mice
0h | 0.5h | 2h | Area under the curve of blood glucose | |
The A Normal group | 8.35±1.64 | 12.48±1.77 | 6.23±1.69 | 19.23±2.77 |
B hyperglycemia model group | 30.03±3.32 | 32.83±0.59 | 30.50±1.82 | 63.22±1.14 |
C medicine matched group | 27.33±3.54 | 31.62±0.76 | 21.75±2.85 | 54.76±2.65 |
E1 Sargassum polypeptide low dose group | 27.53±3.68 | 31.70±1.74 | 30.00±2.94 | 61.08±2.38 |
Dosage group in the E2 Sargassum polypeptide | 26.10±3.94 | 32.53±1.63 | 25.98±3.28 | 58.53±4.34 |
E3 Sargassum polypeptide high dose group | 23.8.±5.72 | 31.43±0.96 | 22.95±3.77 | 54.59±3.86 |
3 conclusions
3.1 the blood fat reducing function of Sargassum polypeptide
Experimental data shows that the Sargassum polypeptide of doses can effectively be controlled the increase of Wistar rat weight; The effect that has the index content such as cholesterol reducing, triglyceride, low-density lipoprotein cholesterol and improve HDL-C content; The also MDA content in the functions for the treatment of hyperlipidemia Wistar rat liver, and increased SOD content.As seen, long-term taking Sargassum polypeptide has certain regulating action to the indices of blood fat, controls the rising of every blood lipids index, can prevent the generation of hyperlipidemia.
3.2 the effect of lowering blood sugar of Sargassum polypeptide
The hyperglycemia model mouse experiment is the result show, the Sargassum polypeptide has certain protective effect to immune organ spleen and thymus, also can suppress the enlargement of liver; Blood sugar increasing had certain inhibitory action, significantly the blood sugar lowering area under curve.As seen, the Sargassum polypeptide has certain auxiliary curative effect aspect the prevention of hyperglycemia and the treatment.
Test example 2
The beverage of embodiment 5 is tried out through tens people, common every day 1~2 time, each 250mL.
Try out 3~6 months phases.
Below lifting several exemplary further specifies.
1, Zhao * *, 45 years old, suffer from hyperlipidemia, often dizzy, tired, drink beverage that the present invention makes after four months, blood lipid level reduces, spirit is clearly better;
2, Lee * *, 52 years old, meet the type ii diabetes diagnostic criteria, two hours blood glucoses are 17.03mmol/L after the meal, drink beverage that the present invention makes after four months, two hours blood glucoses drop to 10.20mmol/L after the meal.
Above-described embodiment is the better embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (7)
1. the purposes of Sargassum polypeptide in preparation blood fat reducing, health-caring product capable of reducing blood sugar, described Sargassum polypeptide adopts following mode to make: the porphyra haitanensis after the immersion and the mixed material of water grind through colloid mill, obtain the porphyra haitanensis serosity, adopt the AS.1398 neutral protease at 45 ℃~55 ℃, pH7.2~7.8, enzyme concentration E/S 8000~12000U.g
-1, substrate bulking value specific concentration 3~7% condition under enzymolysis porphyra haitanensis serosity 5h~10h, obtain enzymolysis solution, with the centrifugal rear collection supernatant of enzymolysis solution, again supernatant is obtained molecular weight less than the active component of 10000 Da through ultra-filtration and separation, for subsequent use after lyophilization or the spray drying.
2. according to claims 1 described purposes, it is characterized in that: preferably adopt the AS.1398 neutral protease at 50 ℃, pH7.5, enzyme concentration E/S 10000U.g
-1, substrate bulking value specific concentration 5% condition under enzymolysis porphyra haitanensis serosity 10h, obtain enzymolysis solution.
3. according to claims 2 described purposes, it is characterized in that: the supernatant of centrifugal rear collection is in one times of the ratio thin up of 1:1, and the supernatant after then will diluting adds ultrafiltration apparatus and separates.
4. blood fat reducing, blood sugar lowering beverage that comprises the Sargassum polypeptide, it is characterized in that: it comprises Sargassum polypeptide and the edible drink formula of blood fat reducing, blood sugar lowering effective dose, described Sargassum polypeptide stock solution adopts following mode to make: the porphyra haitanensis after the immersion and the mixed material of water grind through colloid mill, obtain the porphyra haitanensis serosity, adopt the AS.1398 neutral protease at 45 ℃~55 ℃, pH7.2~7.8, enzyme concentration E/S 8000~12000U.g
-1, concentration of substrate 3~7%(w/v) condition under enzymolysis porphyra haitanensis serosity 5h~10h, obtain enzymolysis solution, with the centrifugal rear collection supernatant of enzymolysis solution, again supernatant is obtained molecular weight less than the Sargassum polypeptide stock solution of the active component of 10000 Da through ultra-filtration and separation.
5. according to claims 4 described blood fat reducing that comprise the Sargassum polypeptide, the blood sugar lowering beverage, it is characterized in that: described drink formula comprises cyclodextrin, malic acid, sodium citrate, sodium isoascorbate, Vc, glycine, white sugar and sodium carboxymethyl cellulose, and according to cyclodextrin 5 ‰~7 ‰, malic acid 0.5 ‰~1 ‰, sodium citrate 0.2 ‰~0.6 ‰, sodium isoascorbate 0.2 ‰~0.6 ‰, Vc 0.2 ‰~0.6 ‰, glycine 0.1 ‰~0.3 ‰, the ratio of white sugar 50 ‰~80 ‰ and sodium carboxymethyl cellulose 0.5~0.8 ‰ adds in the Sargassum polypeptide stock solution.
6. according to claims 5 described blood fat reducing, blood sugar lowering beverages that comprise the Sargassum polypeptide, it is characterized in that: described drink formula also comprises maple sugar essence and roasting barley essence, and adds in the Sargassum polypeptide stock solution according to the ratio of maple sugar essence 0.1 ‰~0.3 ‰ and roasting barley essence 0.5 ‰~1.5 ‰.
7. according to claims 4 described Sargassum polypeptide beverages with blood fat reducing, function of blood sugar reduction, it is characterized in that: the supernatant of centrifugal rear collection is in one times of the ratio thin up of 1:1, and the supernatant after then will diluting adds ultrafiltration apparatus and separates.
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