CN1304548C - Health care wine containing dihydro-myricetin and myricetin composition - Google Patents
Health care wine containing dihydro-myricetin and myricetin composition Download PDFInfo
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- CN1304548C CN1304548C CNB031238882A CN03123888A CN1304548C CN 1304548 C CN1304548 C CN 1304548C CN B031238882 A CNB031238882 A CN B031238882A CN 03123888 A CN03123888 A CN 03123888A CN 1304548 C CN1304548 C CN 1304548C
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- ampelopsin
- wine
- dibydro myricetrin
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- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- LJDZFAPLPVPTBD-UHFFFAOYSA-N nitroformic acid Chemical compound OC(=O)[N+]([O-])=O LJDZFAPLPVPTBD-UHFFFAOYSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
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- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000003495 polar organic solvent Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
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- 239000002594 sorbent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
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Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention provides health-care wine and a preparing method thereof. A composition containing dihydro-myricetin and myricetin, which is used as an additive agent, is contained in the health-care wine. Wine comprises alcohol wine and juice wine. The content of the composition containing dihydro-myricetin and myricetin of the present invention in the wine can be 0.1 to 53%w/v (percentage by weight in volume (gram per liter), the following text is the same), and the range from 2%w/v to the amount of the dissolving saturation of the composition containing dihydro-myricetin and myricetin of the present invention in various kinds of wine is preferable. The composition containing dihydro-myricetin and myricetin of the present invention, which is in the amount, can be directly added into the wine and stirred uniformly to obtain the wine containing the composition containing dihydro-myricetin and myricetin of the present invention, and the composition containing dihydro-myricetin and myricetin is used as the additive agent.
Description
The invention belongs to field of health care products.
Document Walter Karrerr, Birkhauser Verlag, Bassel undstuttgart (1958), P.652, NO:1640 discloses the structural formula of dibydro myricetrin,
But the activity of dibydro myricetrin is not studied.Ampelopsin is a kind of compound known, and its structural formula is:
。Nobody disclosed the composition and use thereof that contains dibydro myricetrin and ampelopsin up to now.
The purpose of this invention is to provide a kind of new composition, it contains dibydro myricetrin and ampelopsin, and wherein the dihydromyricetin cellulose content is 20% to 98% weight ratio, and ampelopsin content is 2% to 80% weight ratio.Can also contain one or more in the present composition and be selected from following composition: arbutin, gallic acid and/or tea-polyphenol.The content of arbutin preferably accounts for the 0-50wt% of dibydro myricetrin and ampelopsin gross weight, the content of gallic acid preferably accounts for the 0-50wt% of dibydro myricetrin and ampelopsin gross weight, and the content of tea-polyphenol preferably accounts for the 0-50wt% of dibydro myricetrin and ampelopsin gross weight.
The applicant is surprisingly found out that the composite health product field of containing dibydro myricetrin and ampelopsin has purposes widely.
The composition that contains dibydro myricetrin and ampelopsin of the present invention has hypoglycemic and reducing blood lipid, function with prevention and/or recovery alcoholic liver injury, can be used as additive is added in various types of wine, itself and wine are complementary, and can not change original colourity and the clarity and the sense of taste of wine.
Another object of the present invention provides the method for compositions that preparation the present invention contains dibydro myricetrin and ampelopsin.Composition of the present invention can also can extract from ampelopsis by described constituent monomers is mixed in proportion preparation.Described ampelopsis comprises ampelopsis cantoniensis, a leaf beautiful grape, Ampelopsis grossedentata, Cayratia japonica (Thunb.) Gagnep., radix ampelopsis, Northeastern Caulis seu folium ampelopsis brevipedunculatae and Vitis Amurensis.Described extracting method can be: decoct vitaceae by all kinds of SOLVENTS (water and/or organic solvent), decoct one or many, decoction liquor is through concentrating cooling, standing over night, detection precipitates the effective component with supernatant, gets to have active precipitation and/or obtain the present composition after further silica gel adsorption.Decoct used solvent preferably water, alcohols, ester class, ketone, ethers and strong polar organic solvent, more preferably low-level chain triacontanol, low-grade fatty acid ester has ketone and ether and/or water, the most preferably methyl alcohol of 3-12 carbon atom, ethanol, propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, sec-butyl alcohol, the trimethyl carbinol, ethyl acetate, methyl acetate, ethyl formate, acetone, methyl ethyl ketone, ether, methyl ethyl ether, methyl tertiary butyl ether, methylene dichloride, chloroform, tetracol phenixin, methyl-sulphoxide, N, dinethylformamide and/or water or the like.Separating the sorbent material that uses can be silica gel.
The evidence composition that contains dibydro myricetrin and ampelopsin of the present invention has hypoglycemic and reducing blood lipid, has the function of prevention and/or recovery alcoholic liver injury, has liver protecting, the function of enhancing immunity systemic immunity power.The composition that contains dibydro myricetrin and ampelopsin of the present invention can be used as additive and is added in various types of wine, makes health promoting wine.An object of the present invention is to provide a kind of composition that contains dibydro myricetrin and ampelopsin as wine of additive and preparation method thereof.Wine comprises alcohol wine and fruit juice wine.The content that contains the composition of dibydro myricetrin and ampelopsin in the wine can be 0.1-53%w/v (percent weight in volume (grams per liter), hereinafter identical), preferably 2%w/v is to the amount of the degree of dissolution saturation of the composition that contains dibydro myricetrin and ampelopsin in various wine.Can be directly the composition that contains dibydro myricetrin and ampelopsin of described amount be added in the wine and stirs, make the composition that contains dibydro myricetrin and ampelopsin wine as additive.Another object of the present invention provides the composition that contains dibydro myricetrin and ampelopsin as the purposes of additive in the various wine of preparation.
Describe the present invention in detail by the following examples.But should be appreciated that these embodiment just illustrate the present invention, rather than in office where face limits the scope of the invention.
Embodiment 1: prepare the present composition from dibydro myricetrin and ampelopsin crystal
Get dibydro myricetrin 80g, ampelopsin 20g gets mixture after said components is mixed in mixing machine.Perhaps get dibydro myricetrin 98g, ampelopsin 2g gets mixture after said components is mixed in mixing machine.Perhaps get dibydro myricetrin 20g, ampelopsin 80g gets mixture after said components is mixed in mixing machine.
Embodiment 2: prepare the present composition from dibydro myricetrin and ampelopsin crystal
Get dibydro myricetrin 20g, ampelopsin 80g, arbutin 2.5g, gallic acid 2.5g, tea-polyphenol 2.5g, said components in mixing machine, mix mixture.Perhaps get dibydro myricetrin 98g, ampelopsin 2g, arbutin 2.5g, gallic acid 2.5g, tea-polyphenol 2.5g, said components in mixing machine, mix mixture.Perhaps get dibydro myricetrin 85g, ampelopsin 15g, arbutin 25g, gallic acid 2g, said components in mixing machine, mix mixture.Perhaps get dibydro myricetrin 50g, ampelopsin 50g, arbutin 25g, tea-polyphenol 2.5g, said components in mixing machine, mix mixture.Perhaps get dibydro myricetrin 70g, ampelopsin 30g, arbutin 5g, said components in mixing machine, mix mixture.
Embodiment 3: prepare the present composition from dibydro myricetrin and ampelopsin crystal
Get first kind of composition (or the third composition of embodiment 2) 30g of embodiment 1 respectively, lactose 53g, Xylo-Mucine 1.5g, Magnesium Stearate 0.5g, 50% ethanol 5ml, said components is mixed the back compacting in flakes in mixing machine.
Embodiment 4: prepare the present composition from dibydro myricetrin and ampelopsin crystal
Get dibydro myricetrin 115g, ampelopsin 15g, Icing Sugar 345g, dextrin 145g, ethanol 5ml, said components is mixed after drying, obtains electuary.
Embodiment 5: prepare the present composition from porcelain ampelopsis
Get the dry young stem and leaf 2kg of Ampelopsis grossedentata, with 95% ethanol (medical ethanol) refluxing extraction twice, each 2 hours, filter, merging filtrate is evaporated to dried, add 1200 milliliters of 85 ℃ of hot water, medicinal extract is fully dissolved, elimination black insolubles, add silica gel (100 order) 200g in the filtrate, filtrate is heated to 60 ℃, take advantage of heat filtering, filtrate is cooled to room temperature, precipitation, standing over night, obtain yellow mercury oxide, productive rate: 11.2%.Dihydromyricetin cellulose content 74.8% weight ratio wherein, ampelopsin content 21.1% weight ratio, the content of arbutin is 2.8% weight ratio, the content of gallic acid is 1.3% weight ratio.Thin-layer chromatographic analysis polymeric amide thin-layer chromatography, specification 8 * 8cm, developping agent: methyl alcohol, developer: 4% iron trichloride-ethanolic soln, R
The f gallic acid=0.72, R
The f arbutin=0.54, R
The f dibydro myricetrin=0.50, R
The f ampelopsin=0.21.
Perhaps get the dry young stem and leaf 2kg of Ampelopsis grossedentata, with 95% ethanol (medical ethanol) refluxing extraction twice, each 2 hours, filter, merging filtrate is evaporated to dried, add 1200 milliliters of 85 ℃ of hot water, medicinal extract is fully dissolved, elimination black insolubles, filtrate is used twice of equal-volume ethyl acetate extraction, combined ethyl acetate solution, be evaporated to driedly, obtain yellow mercury oxide, productive rate: 10.25%.Dihydromyricetin cellulose content 76% weight ratio wherein, ampelopsin content 18% weight ratio, the content of arbutin is 3.0% weight ratio, and the content of gallic acid is 2% weight ratio, and all the other are R in the porcelain ampelopsis
F5=0.41 composition.Thin-layer chromatographic analysis polymeric amide thin-layer chromatography, specification 8 * 8cm, developping agent: methyl alcohol, developer: 4% iron trichloride-ethanolic soln, R
The f gallic acid=0.72, R
The f arbutin=0.54, R
The f dibydro myricetrin=0.50, R
The f ampelopsin=0.21, R
F5=0.41.
Embodiment 6: prepare the present composition from the Northeastern Caulis seu folium ampelopsis brevipedunculatae plant
Under the room temperature, the Northeastern Caulis seu folium ampelopsis brevipedunculatae 1000 gram flush away dust with collecting add 4000 ml waters, supersound extraction 0.5 hour is filtered, and residue adds 1000 milliliters of propyl alcohol again, supersound extraction is 0.5 hour again, refilters, and filtrate merges, be concentrated into 2 milliliters, add 500 milliliters of 100 ℃ of hot water, be cooled to 60 ℃, leave standstill, filter.Filtrate is heated to 70 ℃ with silica gel (100 order) 100g absorption with filtrate, takes advantage of heat filtering, and filtrate is cooled to room temperature, precipitation, and standing over night obtains yellow mercury oxide, productive rate: 10.3%.Dihydromyricetin cellulose content 58% weight ratio wherein, ampelopsin content 14% weight ratio, the content of arbutin is 1.4% weight ratio, and the content of gallic acid is 1.7% weight ratio, and all the other are R in the Northeastern Caulis seu folium ampelopsis brevipedunculatae plant
F5=0.43 composition.Thin-layer chromatographic analysis polymeric amide thin-layer chromatography, specification 8 * 8cm, developping agent: methyl alcohol, developer: 4% iron trichloride-ethanolic soln, R
The f gallic acid=0.76, R
The f arbutin=0.55, R
The f dibydro myricetrin=0.54, R
The f ampelopsin=0.22, R
F5=0.43.
The example 7 hypoglycemic tests of composition that contain dibydro myricetrin and ampelopsin of the present invention
1. materials and methods
1.1. tried thing: first kind of composition that embodiment 5 obtains
1.2. experimental animal and testing conditions
Select the female Kunming mouse of healthy cleaning level for use, body weight 24 ± 2g, animal-feed operative norm GB14924-94.Sense environmental conditions, temperature range 20-25 ℃, relative humidity scope 40-70%.
1.3. dosage is selected
Test divides intact animal and two batches of hyperglycemia model animals to carry out, and respectively establishes 1 control group (distilled water) and pool give birth to board blood pure 0.25,0.50,1.50g/kg dosage group, is equivalent to 5,10 and 30 times of people's recommended intake respectively, and filling stomach amount is 0.1ml/10g.
1.4. key instrument and reagent
Tetraoxypyrimidine (Alloxan): Sigma company; 50% glucose injection: new Cao in Yancheng pharmaceutical factory.
The super blood sugar detection instrument of SUPER GLUCOCARD II GT-1640 type: Japanese capital person first science Co., Ltd. produces: the GLUCOCARD blood sugar test paper: Japanese capital person first science Co., Ltd. produces.
1.5. experimental technique
1.5.1. intact animal: fasting was got mouse and is surveyed blood sugar after 5 hours.40 of the mouse of screening fasting plasma glucose in normal range are divided into four groups at random by glucose level, 10 every group, are respectively each dosage group of normal control group and sample (difference is not more than 1.1mmol/L between group) "
1.5.2. hyperglycemia model animal: strict fasting is after 24 hours, mouse tail vein injection tetraoxypyrimidine 50mg/kgBW, and fasting is 5 hours after 6 days, surveys blood sugar.The mouse 40 of screening fasting blood sugar＞10mmol/L is done, and is divided into four groups at random by glucose level, 10 every group, is respectively each dosage group of model control group and sample (difference is not more than 1.1mmol/L between group).
1.5.3. the mensuration of fasting plasma glucose and sugar tolerance: by design dosage mouse continuous irrigation stomach was tried thing 30 days, fasting is 5 hours then, gives glucose 1.5g/kgBW and irritates stomach, measures blood glucose value respectively in 0,0.5,2.0 hour 3 phase.
1.6. statistical method
All experimental data adopts SPSS/PC software package (one-way analysis of variance) to handle on microcomputer.
2. result
2.1. contain of the influence of the composition of dibydro myricetrin and ampelopsin to the normal mouse body weight
Table 1 respectively organize normal mouse initial body weight, mid-term body weight, finish body weight
| Group | Initial body weight | Mid-term body weight | Finish body weight |
| Number of animals | Body weight (g) | Number of animals | Body weight (g) | Number of animals | Body weight (g) | ||
| Normal control | 10 | 23.4±1.0 | 10 | 30.5±1.5 | 10 | 32.4±1.6 | |
| Tried thing | 0.25g/kg | 10 | 23.7±1.0 | 10 | 31.3±1.3 | 10 | 33.0±1.7 |
| 0.50g/kg | 10 | 23.6±1.1 | 10 | 30.8±1.7 | 10 | 32.2±1.8 | |
| 1.50g/kg | 10 | 23.6±1.3 | 10 | 31.0±1.4 | 10 | 33.7±1.7 | |
| The F value | 0.115 | 0.475 | 0.353 | ||||
| The P value | 0.951 | 0.701 | 0.787 | ||||
Annotate: tried each dosage group mouse body weight of thing and compare the equal nonsignificance of difference (variance analysis, P＞0.05) in each trial period and normal control group
2.2. contain of the influence of the composition of dibydro myricetrin and ampelopsin to the normal mouse fasting plasma glucose
Table 2 contains the influence of the composition of dibydro myricetrin and ampelopsin to the normal mouse fasting plasma glucose
| Group | Number of animals (only) | Fasting blood sugar (mmol/L) | Difference | ||
| Before the test | After the test | ||||
| Normal control | 10 | 4.7±0.5 | 4.6±0.7 | 0.1±06 | |
| Tried thing | 0.25g/kg | 10 | 4.4±0.7 | 4.3±0.8 | 0.1±1.2 |
| 0.50g/kg | 10 | 4.6±0.9 | 4.4±0.9 | 0.2±1.0 | |
| 1.50g/kg | 10 | 4.5±0.8 | 4.5±0.8 | 0.0±0.5 | |
| The F value | 0.388 | 0.302 | 0.099 | ||
| The P value | 0.762 | 0.82 | 0.960 | ||
Annotate: tried each dosage group fasting blood sugar of thing and normal control group relatively, difference there are no significant meaning (variance analysis, P＞0.05).
2.3. contain of the influence of the composition of dihydromyricetin rope and ampelopsin to the normal mouse postprandial blood sugar
The composition of table 3 dibydro myricetrin and ampelopsin is to the influence of normal mouse postprandial blood sugar
| Group | Number of animals (only) | Glucose (g/kgBW) | 0h | Blood glucose value (mmol/L) | ||
| 0.5h | 2h | |||||
| Normal control | 10 | 1.5 | 4.6±0.7 | 10.4±1.2 | 5.5±0.7 | |
| Tried thing | 0.25g/kg | 10 | 1.5 | 4.3±0.8 | 10.8±1.0 | 5.2±1.0 |
| 0.50g/kg | 10 | 1.5 | 4.4±0.9 | 10.5±1.6 | 5.0±1.2 | |
| 1.50g/kg | 10 | 1.5 | 4.5±0.8 | 10.5±1.4 | 5.2±1.9 | |
| The F value | 0.302 | 0.230 | 0.529 | |||
| The P value | 0.824 | 0.875 | 0.665 | |||
Annotate: tried each dosage group postprandial plasma glucose level of thing and normal control group relatively, difference there are no significant meaning (variance analysis, P＞0.05).
2.4. contain of the influence of the composition of dibydro myricetrin and ampelopsin to the normal mouse sugar tolerance
The composition of table 4 dibydro myricetrin and ampelopsin is to the influence of normal mouse sugar tolerance
| Group | Number of animals (only) | Blood sugar increasing and reduction amplitude (mmol/L) | ||
| 0.5h-0h | 0.5h-2h | |||
| Normal control | 10 | 5.8±0.9 | 4.8±0.9 | |
| Tried thing | 0.25g/kg | 10 | 6.5±0.6 | 5.6±1.2 |
| 0.50g/kg | 10 | 6.1±1.0 | 5.5±1.0 | |
| 1.50g/kg | 10 | 6.0±1.4 | 5.3±1.3 | |
| The F value | 0.998 | 1.018 | ||
| The P value | 0.405 | 0.396 | ||
Annotate: tried each dosage group sugar tolerance value of thing and normal control group relatively, difference there are no significant meaning (variance analysis, P＞0.05).
2.5. the composition that contains dibydro myricetrin and ampelopsin is to the influence of the hyperglycemia mouse body weight that causes
Table 5 respectively organize the hyperglycemia model mouse initial body weight, mid-term body weight, finish body weight
| Group | Initial body weight | Mid-term body weight | Finish body weight | ||||
| Number of animals | Body weight (g) | Number of animals | Body weight (g) | Number of animals | Body weight (g) | ||
| Normal control | 10 | 24.3±1.3 | 10 | 30.7±2.0 | 10 | 31.2±1.9 | |
| Tried thing | 0.252g/kg | 10 | 24.5±2.7 | 10 | 31.8±2.2 | 10 | 32.4±2.8 |
| 0.50g/kg | 10 | 24.0±1.4 | 10 | 31.7±2.9 | 10 | 32.6±2.9 | |
| 1.50g/kg | 10 | 23.8±1.5 | 10 | 30.9±1.9 | 10 | 30.8±1.7 | |
| The F value | 0.271 | 0.576 | 1.418 | ||||
| The P value | 0.846 | 0.634 | 0.253 | ||||
Annotate: tried each stage body weight of each dosage group of thing and model control group relatively, difference there are no significant meaning (variance analysis, P＞0.05).
2.6. the composition that contains dibydro myricetrin and ampelopsin is to the influence of the hyperglycemia mouse fasting plasma glucose that causes
The composition of table 6 dibydro myricetrin and ampelopsin is to the influence of hyperglycemia model mouse fasting plasma glucose
| Group | Number of animals (only) | Fasting blood sugar (mmol/L) | Difference | ||
| Before the test | After the test | ||||
| Normal control | 10 | 22.9±3.3 | 20.4±5.1 | 2.5±4.2 | |
| Tried thing | 0.25g/kg | 10 | 22.9±3.0 | 13.2±4.1* | 9.7±3.7* |
| 0.50g/kg | 10 | 22.4±2.5 | 15.3±6.4 | 7.2±4.7 | |
| 1.50g/kg | 10 | 22.5±3.5 | 17.1±6.3 | 5.4±6.1 | |
| The F value | 0.068 | 3.058 | 4.137 | ||
| The P value | 0.977 | 0.041 | 0.0123 | ||
Annotate: compare * P＞0.05 (q check) with model control group
Tried thing low dose group test back fasting blood sugar and front and back fasting plasma glucose difference and model control group relatively, difference all has significantly (variance analysis, P＞0.05).
2.7. the composition that contains dibydro myricetrin and ampelopsin is to the influence of the hyperglycemia mouse postprandial blood sugar that causes
The composition of table 7 dibydro myricetrin and ampelopsin is to the influence of hyperglycemia type mouse postprandial blood sugar
| Group | Number of animals (only) | Glucose (g/kgBW) | 0h | Blood glucose value (mmol/L) | ||
| 0.5h | 2h | |||||
| Normal control | 10 | 1.5 | 20.4±5.1 | 31.2±2.7 | 24.7±3.7 | |
| Tried thing | 0.25g/kg | 10 | 1.5 | 13.2±4.1* | 26.4±4.2 | 16.5±6.6* |
| 0.50g/kg | 10 | 1.5 | 15.3±6.4 | 27.1±5.7 | 16.9±7.2* | |
| 1.50g/kg | 10 | 1.5 | 17.1±6.3 | 28.4±4.3 | 20.2±7.2 | |
| The F value | 3.058 | 2.393 | 3.536 | |||
| The P value | 0.041 | 0.085 | 0.024 | |||
Annotate: tried the low agent group of thing after the meal the 0h blood glucose value and low in the dosage group after the meal the 2h blood glucose value compare with model control group, difference all has significantly (variance analysis, P＞0.05).
2.8. the composition that contains dibydro myricetrin and ampelopsin is to the influence of the hyperglycemia mouse sugar tolerance that causes
The composition of table 8 dibydro myricetrin and ampelopsin is to the influence of hyperglycemia model mouse sugar tolerance
| Group | Number of animals (only) | Blood sugar increasing and reduction amplitude (mmol/L) | ||
| 0.5h-0h | 0.5h-2h | |||
| Normal control | 10 | 10.8±3.2 | 6.5±2.9 | |
| Tried | 0.25g/kg | 10 | 13.3±1.7 | 9.9±3.0 |
| 0.50g/kg | 10 | 11.8±3.6 | 10.1+3.7 | |
| 1.50g/kg | 10 | 11.3±2.9 | 8.3±4.2 | |
| The F value | 1.301 | 2.255 | ||
| The P value | 0.289 | 0.099 |
Annotate: tried each dosage group sugar tolerance value of thing and model control group relatively, difference all has significantly (variance analysis, P＞0.05).
Experimental result shows that the composition of dibydro myricetrin and ampelopsin has the blood sugar regulation effect.
Carry out identical test with the second kind of composition of the third composition of first kind of composition of embodiment 1, embodiment 2, embodiment 5 or the composition of embodiment 6, obtained roughly the same result.
Embodiment 8: the test that contains the composition reducing blood-fat of dibydro myricetrin and ampelopsin of the present invention
Animal is divided into 4 groups at random, normal control group, positive controls, model control group, extracting solution group.Every group of 10 mouse, once-a-day, successive administration 10 days, the normal control group is not given any medicine.All the other all to high lipoprotein emulsion 0.5ml/ only respectively organize mouse, form experimental hyperlipidemia, overnight fasting after medication in 10 days, press enzyme process and detect serum total cholesterol (TC), triglyceride level (TG), high density lipoprotein cholesterol (HDL-C) content from the mouse orbit extracting vein blood next day.The result shows, model control group serum TC, TG value obviously raise, the HDL-C value descends, compare with model, first kind of composition group of embodiment 5 can obviously reduce serum TC, TG value, HDL-C is slightly raise, illustrate that this extract can suppress the mouse blood fat rising that high lipoprotein emulsion causes, has effect for reducing fat.Carry out identical test with the second kind of composition of the third composition of first kind of composition of embodiment 1, embodiment 2, embodiment 5 or the composition of embodiment 6, obtain identical result.
1. material and method
1.1. tried thing: first kind of composition that embodiment 5 obtains is for experiment with the turbid solution that distilled water is mixed with desired concn trying thing.
1.2. laboratory animal: SPF level Wistar rat, body weight 170-190g, female, totally 55 (5 are standby).Experimental temperature: 23-25 ℃, relative humidity: 65-70%.
1.3. feed:
1.3.1. common basal feed: prescription slightly.(the pure mouse material of the Wistar of Hubei Province Preventive Medicine Academy's Experimental Animal Center preparation).
1.4. the dosage grouping: experiment is divided into five groups, be normal control group, hyperlipidemia model control group and tried basic, normal, high three the dosage groups of thing, dosage is respectively 0.5,1.0,1.5g/kg.bw, is equivalent to manufacturer and recommends 10,20,30 times of human body daily intaking amount 3g/60kg.bw.
1.5. experimental technique:
Rat adaptability is raised and is begun experiment after 3 days.Get rat tail blood, measure serum total cholesterol (TC0), triglyceride (TG0), high density lipoprotein cholesterol (HDL-c0) basic value.According to the TC0 level, rat is divided into five groups at random, every group of 10 animals, promptly basic, normal, high dosage group, normal control group and hyperlipidemia model control group.Except that the normal control group gives the basal feed, all the other each groups all give high lipid food.Three are tried agent amount group and irritate stomach by the capacity per os of 1.0ml/100g.bw and give, the feed of freely drinking water of normal control group and hyperlipidemia model control group.Give 30 days continuously, detected TC, TG, HDL-C value on the 30th day in irritating stomach.
1.6. key instrument and reagent: SABA-18 type automatic clinical chemistry analyzer (Italy produces), standard quality controlled serum and corresponding reagent box are produced by Shanghai Foxing Changzheng medical science Co., Ltd.
1.7. data processing: adopt the STATA statistical software to carry out statistical study
2. result
2.1. first kind of composition is to the influence of hyperlipidemia model the weight of animals: as seen from Table 9, compare with model control group, basic, normal, high dosage group around the time rat body weight all be lower than model control group, basic, normal, high dosage group and model control group comparing difference have significance (P＜0.01, P＜0.01, P＜0.01).With the normal control group relatively, body weight all is higher than the normal control group around the basic, normal, high dosage group the, but credit is analysed by statistics, difference does not have significance.
Table 9 contains the composition of dibydro myricetrin and ampelopsin
To the influence of high blood lipid model rat body weight (g, x ± s)
| Dosage g/kg.bw | Number of animals (only) | Body weight | ||||
| Starting weight | First week | Second week | The 3rd week | Heavy eventually | ||
| The normal control model contrasts 0.5 1.0 1.5 | 10 10 10 10 10 | 181.4±2.8 181.7±3.5 182.4±4.1 183.1±2.8 182.3±2.5 | 202.2±6.2 208.7±2.6 208.1±2.6 207.0±2.5 208.3±2.8 | 219.6±4.7 224.5±4.4 221.7±3.4 221.5±2.4 220.3±3.2 | 226.2±3.7 231.4±3.5 227.9±3.4 227.7±2.0 226.2±3.8 | 230.7±4.0** 237.6±3.7 233.6±3.0** 233.2±2.1** 232.7±3.1** |
* and model control group be P＜0.01 relatively
2.2. first kind of composition is to the influence of rat total cholesterol level: as seen from Table 10, compare with model control group, middle and high dosage group total cholesterol level descended obviously at experimental session when experiment finished, and credit is analysed by statistics, and difference has significance (p＜0.05, P＜0.01).
Table 10 contains the composition of dibydro myricetrin and ampelopsin
To the influence of rat total cholesterol level (mmol/L, x ± s)
| Dosage g/kg.bw | Number of animals (only) | Total cholesterol | |||
| Before the test | The P value | After the test | The P value | ||
| The contrast of normal control model | 10 10 | 1.80±0.07 1.81±0.11 | 0.786 - | 1.81±0.16 3.72±0.34 | 0.000 - |
| 0.5 1.0 1.5 | 10 10 10 | 1.83±0.10 1.82±0.10 1.84±0.11 | 0.803 0.928 0.634 | 3.35±0.55 3.10±0.54* 2.86±0.57** | 0.269 0.010 0.001 |
* compare P＜0.05 with model control group, * * and model control group be P＜0.01 relatively
2.3. first kind of composition is to the influence of rat content of triglyceride: as seen from Table 11, compare with model control group, basic, normal, high dosage group content of triglyceride descends obviously during off-test, and credit is analysed by statistics, and difference has significance (P＜0.05, P＜0.01, P＜0.01).
Table 11 contains the composition of dibydro myricetrin and ampelopsin
To the influence of rat content of triglyceride (mmol/L, x ± s)
| Dosage g/kg.bw | Number of animals (only) | Total cholesterol | |||
| Before the test | The P value | After the test | The P value | ||
| The normal control model contrasts 0.5 1.0 1.5 | 10 10 10 10 10 | 1.24±0.25 1.26±0.27 1.23±0.19 1.24±0.18 1.21±0.15 | 0.828 - 0.788 0.820 0.584 | 1.29±0.11 2.30±0.47 1.90±0.29* 1.89±0.24** 1.86±0.12** | 0.000 - 0.011 0.009 0.008 |
* compare P＜0.05 with model control group, * * and model control group be P＜0.01 relatively
2.4. a composition is to the influence of rat high density lipoprotein cholesterol content: as seen from Table 12, compare with model control group, basic, normal, high dosage group has the rising effect to high density lipoprotein cholesterol, and credit is analysed by statistics, and difference has significance (P＜0.05).
Table 12 contains the composition of dibydro myricetrin and ampelopsin
To the influence of rat high density lipoprotein cholesterol content (mmol/L, x ± s)
| Dosage g/kg.bw | Number of animals (only) | High density lipoprotein cholesterol | |||
| Before the test | The P value | After the test | The P value | ||
| The normal control model contrasts 0.5 1.0 1.5 | 10 10 10 10 10 | 1.13±0.12 1.12±0.18 1.10±0.08 1.13±0.09 1.14±0.05 | 0.872 - 0.644 0.841 0.733 | 1.06±0.11 0.84±0.11 0.94±0.10* 0.93±0.07* 0.95±0.08* | 0.000 - 0.027 0.042 0.015 |
* compare P＜0.05 with model control group
2.5. first kind of composition is to the influence of rat fat level: this experimental technique is a high lipid food and tried thing and give simultaneously, belongs to preventative, so TG, TC decline and HDL-c rising value all with the comparison of hyperlipidemia model control group.By table 13 as seen, middle and high dosage group total cholesterol level fall is respectively 16.7%, 23.1%, basic, normal, high dosage group content of triglyceride fall is respectively 17.4%, 17.8%, 19.1%, and basic, normal, high dosage group high density lipoprotein cholesterol content lift-off value is 4.64,5.03,4.26mg/dl.
Table 13 contains the influence of the composition of dibydro myricetrin and ampelopsin to the rat fat level
| Dosage g/kg.bw | Number of animals (only) | TC (％) | TG (％) | HDL-C (mg/dl) |
| 0.5 1.0 1.5 judging criterions: | 10 10 10 | 10.3 16.7 23.1 descend＞10% | 17.4 17.8 19.1 descend＞15% | 4.64 5.03 4.26 rising 4mg/dl |
Annotate: HDL-c lift-off value (mg/dl) is mmol/L * 38.7=mg/dl conversion result
Carry out identical test with the second kind of composition of the third composition of first kind of composition of embodiment 1, embodiment 2, embodiment 5 or the composition of embodiment 6, obtained roughly the same result.
Embodiment 9: the present invention contains the composition prevention of dibydro myricetrin and ampelopsin and/or recovers the function of alcoholic liver injury
1, materials and methods
1.1 test product: the present invention contains the composition of dibydro myricetrin and ampelopsin, sees first kind of composition of embodiment 5.
1.2 laboratory animal: bull mouse (18-22 gram), 10 every group.
1.3 experimental technique and step
1.3.1 dosage grouping and given the test agent give the time
Experiment is established blank group, model control group and is subjected to three dosage groups of test product (to be respectively 10mg/kg, 20mg/kg, 40mg/kg, prepare with dehydrated alcohol respectively, concentration is respectively 0.5mg/ml, 1.0mg/ml, 2.0mg/ml), cause liver injury model with dehydrated alcohol (analytical pure), dehydrated alcohol concentration is 50% (with distilled water diluting), mouse stomach 12-14ml/kgBW (amounting to alcoholic acid dosage is 6000-7000mg/kgBW).It is 30 days that given the test agent gives the time.
1.3.2 give the approach of given the test agent
Per os is irritated stomach and is given given the test agent, and writes down feed intake or the amount of drinking water of every animal.
1.3.3 experimental procedure
Be subjected to three dosage groups of test product per os every day to irritate the ethanolic soln 0.2ml that stomach gives given the test agent, blank group and model control group give distilled water.Animal is weighed weekly twice.When giving given the test agent and finishing model control group and each sample sets are once irritated stomach and give 50% ethanol 12ml/kg BW, the blank group is given distilled water, and fasting was put to death animal in 15 hours, carried out the detection and the histopathologic examination of every index.
1.3.4 detection index
Mda in the hepatic tissue (MDA) reduced glutathion (GSH)
The content of triglyceride level (TG)
1.4 lipid peroxide degraded product mda (MDA) measuring method in the liver homogenate
1.4.1 principle
MDA (malondiadehycle) is one of snperoxiaized end product of cytolemma lipid, but detect the degree of its content indirect Estimation lipid peroxidation, MDA and thiobarbituricacid are warm altogether under acidic conditions, form pink, absorption peak is at 535nm, and this can record the content of MDA tool.
1.4.2 instrument and reagent
Instrument product 721 spectrophotometers, micro sample-adding product, thermostat water bath, generic centrifuge, DL device, tool plug centrifuge tube, tissue homogenizer
Reagent 0.2M acetate buffer PH3.5
0.2M acetic acid solution 185ml
0.2M sodium acetate solution 15ml
1mmol/L tetraethoxypropane (stock solution was preserved 3 months for 4 ℃) faces with before being diluted with water to 40nmol/ml
8.1% sodium lauryl sulphate SDS
0.8% thiobarbituricacid TBA
0.2M phosphate buffered saline buffer PH7.4
0.2M Sodium phosphate dibasic 1920ml
0.2M potassium primary phosphate 480ml
1.4.3 experimental procedure
1.4.3.1 specimen preparation
Tissue homogenate sample: get a certain amount of required internal organs, normal saline flushing, wipe away dried, weigh, shred, put in the homogenizer, add the 0.2M phosphate buffered saline buffer, spare 10s with 2000r/min, intermittently 30s, carry out repeatedly 3 times, it is even with (W/V) to make 5% tissue, the centrifugal 5 ~ 10min of 3000r/min, and it is to be measured to get supernatant liquor.
1.4.3.2 sample determination
Table 14
| Reagent | Blank pipe | Sample hose | Standard pipe |
| 5% tissue homogenate 40nmol/ml tetraethoxypropane 8.1%SDS 0.2M acetate buffer 0.8%TBA H2O | 0.2ml 1.5ml 1.5ml 0.8ml | 0.1ml 0.2ml 1.5ml 1.5ml 0.7ml | 0.1ml 0.2ml 1.5ml 1.5ml 0.7ml |
| Mixing, lucifuge boiling water bath 60min, the flowing water cooling is in the 532nm colorimetric | |||
1.4.3.3 calculate
A: blank pipe absorbancy
B: the glimmering absorbancy of sample
F: tetraethoxypropane absorbancy
C: tetraethoxypropane concentration
K: extension rate
1.4.3.4 data processing and result judge
The The data variance analysis, but need to carry out homogeneity test of variance earlier by the program of variance analysis, variance is neat, calculates the F value, F value/F0.05, and conclusion: each organizes the mean differences does not have significance; F value 〉=F0.05, P≤0.05 is added up with the comparative approach in twos of a plurality of experimental group and a control group mean; The data of abnormal or heterogeneity of variance are carried out the conversion of suitable variable, wait to satisfy normal state or variance are neat require after, add up with the data after changing; If do not reach normal state or the neat purpose of variance yet after the variable conversion, use rank test instead and add up.
The result judges
Under the prerequisite that model is set up, the MDA content of given the test agent group and model control group compare, and difference has significance, judges this index positive as a result.
1.5. liver homogenate reduced glutathion (GSH) measuring method
1.5.1 principle
GSH and 5,5 '-two sulphur are reflected at nitro formic acid (DTNB) can generate xanchromatic 5-sulfo-2-nitro formic acid positively charged ion under the GSH-Px catalysis, in the 423nm wavelength maximum absorption band is arranged, and measures this ionic concn, can calculate the content of GSH.
1.5.2 reagent
0.9% physiological saline
4% sulphosalicylic acid solution
0.1mol/L PBS solution (pH=8.0)
Na
2HPO
4 13.452g
KH
2PO
4 0.722g
Adding distil water is to 1000ml
0.004%DTNB solution: take by weighing DTNB 40mg and be dissolved in the 0.1mol/L PBS solution (pH=8.0) of 1000ml.
Folded fluorine is received damping fluid
NaN
3 16.25mg
EDTA-
Na2 7.44mg
Na
2HPO
4 1.732g
NaH
2PO
4 1.076g
Adding distil water is transferred pH7.0,4 ℃ of preservations to 1000ml with small amount of H Cl, NaOH.
Standardized solution: take by weighing reduced form GSH15.4mg, add nitrine and receive damping fluid to 50ml, final concentration is 1mmol/L
1.5.3 method
1.5.3.1 sample determination: get liver 0.5g and add physiological saline 5ml and fully grind to form screened stock (10% liver homogenate), get slurries 0.5ml behind the mixing and add 4% sulphosalicylic acid 0.5ml mixing, 3000rpm is centrifugal 10 minutes under the room temperature, gets supernatant liquor and is sample.
Table 15
| Measure pipe | Blank pipe | |
| Sample 4% sulphosalicylic acid DTNB | 0.5ml - 4.5ml | - 0.5ml 4.5ml |
Mixing, room temperature were placed after 10 minutes, and the 412nm place measures absorbancy.
1.5.3.2 typical curve
Table 16
| 1 | 2 | 3 | 4 | 5 | 6 | |
| 1mmol/L (ml) physiological saline (ml) DTNB (ml) GSH measures (μ mol/L) | 0 0.50 4.50 0 | 0.05 0.45 4.50 100 | 0.10 0.40 4.50 200 | 0.15 0.35 4.50 300 | 0.20 0.20 4.50 400 | 0.25 0.25 4.50 500 |
1.5.3.3 calculate
Sample GSH content (μ mol/g hepatic tissue)=corresponding curve concentration value ((μ mol/L) ÷ 50g/L
1.5.4 data processing and result judge
The The data variance analysis, but the program of demanding party's difference analysis is carried out homogeneity test of variance earlier, and variance is neat, calculates the F value, F value＜F
0.05, conclusion: each organizes the mean differences does not have significance; The data of abnormal or heterogeneity of variance are carried out the conversion of suitable variable, wait to satisfy normal state or variance are neat require after, add up with the data after changing; If do not reach normal state or the neat purpose of variance yet after the variable conversion, use rank test instead and add up.
The result judges
Under the prerequisite that model is set up, the reduced form GSH content of given the test agent group and model control group compare, and difference has significance, judges this index positive as a result.
1.6 triglyceride in the liver homogenate (TG) measuring method
1.6.1 measuring method: adopt the triglyceride content in triglyceride mensuration test kit (Phosphoric acid glycerol esters oxydase peroxidase method) mensuration 10% liver homogenate.Identical with the serum levels of triglyceride measuring method, to operate by operation instructions with equivalent 10% liver homogenate alternative serum, measurement result is heavily represented with the mmol/g liver.
1.6.2 data processing and result judge
The The data variance analysis, but need to carry out homogeneity test of variance earlier by the program of variance analysis, variance is neat, calculates the F value, F value＜F
0.05, conclusion: each organizes the mean differences does not have significance; F value 〉=F
0.05, P 〉=0.05 is added up with the comparative approach in twos of mean between a plurality of experimental group and control group, and the data of abnormal or heterogeneity of variance are carried out suitable variable conversion, wait to satisfy normal state or variance are neat require after, add up with the data after changing; If do not reach normal state or the neat purpose of variance yet after the variable conversion, use rank test instead and add up.
The result judges
Under the prerequisite that model is set up, the TG of given the test agent group and model control group compare, and difference has significance, judges this index positive as a result.
1.7 the hepatic pathology histology changes, Case definition and result judge
1.7.1 experiment material: do the square section from the left lobe of liver middle part and draw materials, frozen section, Sudan III dyeing.
1.7.2 microscopy:, observe whole tissue slice continuously with 40 times of object lens from the pathological change that one of liver is looked closely wild opening entry cell.Main observation fat drops in distribution, scope and the area of liver.
1.7.3 standards of grading
The liver cell lactones drips and is dispersed in, rare 0 minute
Contain the liver cell that fat drips and be no more than 1/4 1 fens
Contain the liver cell that fat drips and be no more than 1/2 2 fens
Contain the liver cell that fat drips and be no more than 3/4 3 fens
Hepatic tissue is almost dripped by fat and replaces 4 fens
1.7.4 data processing and result judge
Adopt variance analysis, but need to carry out variance agent check earlier by the program of variance analysis, variance is neat, calculate the F value, F value＜F0.05, conclusion: each organizes the mean differences does not have significance: F value 〉=F0.05, and P≤0.05 is added up with the comparative approach in twos of mean between a plurality of experimental group and control group; It is conversion that the data of abnormal or heterogeneity of variance are carried out suitable change, wait to satisfy normal state or variance are neat require after, add up with the data after changing; If do not reach normal state or the neat purpose of variance yet after the variable conversion, use rank test instead and add up.
The result judges
Under the prerequisite that model is set up, between any one dosage group of model control group and given the test agent, steatosis alleviates, and the difference on the statistics is arranged, and can be judged as positive findings.
1.8 the result judges: satisfy arbitrary condition, the decidable given the test agent has has auxiliary protection function to alcoholic liver button internal affairs
1.8.1 liver MDA, reduced form GSH and three of TG detect the index positive as a result.
1.8.2. wantonly two index positives and the histopathologic examination positive as a result in liver MDA, reduced form GSH and three indexs of TG.
2. result
2.1 lipid peroxide degraded product mda (MDA) in the liver homogenate
As shown in Table 17, the damage control group is compared with negative control group, and MDA content obviously raises in the hepatic tissue, and difference has utmost point significance (P＜0.01); Tried each dosage group MDA content of thing and reduced, and utmost point significant difference (P＜0.01) is arranged than the damage control group.Illustrate that being tried thing can reduce MDA content in the hepatic tissue.
2.2 liver homogenate reduced glutathion (GSH)
As shown in Table 17, the damage control group is compared GSH content with the feminine gender group and is obviously reduced, and difference has utmost point significance (P＜0.01).Tried each dosage group GSH content of thing and be higher than the damage control group, difference has significance (P＜0.01).Illustrate and tried the exhaustion that thing can effectively stop GSH.
2.3 triglyceride in the liver homogenate (TG)
As shown in Table 17, damage control group and negative control compare, and liver TG content obviously raises, and difference has utmost point significance (P＜0.01).Tried each dosage group TG content of thing and be starkly lower than the damage control group, difference has significance (P＜0.05).Illustrate and tried the content that thing can reduce TG in the liver.
Table 17 contains the composition of dibydro myricetrin and ampelopsin
Influence to mda, reduced glutathion and triglyceride
| Group | Number of animals | MDA (μ mol/g liver is heavy) | GSH (μ mol/g liver is heavy) | TG (the mmol/g liver is heavy) | |||
| Mean value | The P value | Mean value | The P value | Mean value | The P value | ||
| Control group | 10 | 7.59±0.59 | 0.000 | 15.34±1.26 | 0.000 | 10.08±1.20 | 0.000 |
| Model group | 10 | 17.08±2.30 | 8.89±0.96 | 50.94±1.22 | |||
| Low dose group | 10 | 11.25±1.34 | 0.000 | 11.95±0.99 | 0.000 | 38.32±2.75 | 0.000 |
| Middle dosage group | 10 | 11.65±1.46 | 0.000 | 11.89±1.01 | 0.000 | 32.32±2.15 | 0.000 |
| High dose group | 10 | 12.32±1.39 | 0.001 | 10.93±1.05 | 0.000 | 36.48±2.19 | 0.000 |
2.4 hepatic pathology histological examination: get liver (Zuo Daye) 10% formalin fixed, the fat stains of frozen section Sudan III, haematoxylin redyeing, mounting, microscopy the results are shown in Table 18.
1) negative control treated animal: have 3 examples to see that slight steatosis appears in liver cell.
2) damage control animals: the diffusivity steatosis all appears in liver cell, and fat drips and is classified as +++~ ++ ++ level.
3) middle and high dosage group hepatic cell fattydegeneration degree is starkly lower than positive controls, and difference has significance (P＜0.05).Low dose group hepatic cell fattydegeneration degree and positive controls be there was no significant difference relatively.
Table 18 pair liver fat drips the influence of distribution
| Group (mg/kg) | Number of animals (only) | Total mark | The P value |
| Negative control (0) damage contrast (0) | 10 10 | 3 39## | ＜0.01 |
| Dosage (20) high dosage (40) in the low dosage (10) | 10 10 10 | 28 28* 26* | ＜0.05 ＜0.05 |
Compare with negative control group: ##P＜0.01: compare with the damage control group: * P＜0.05
3.. conclusion:
Test-results shows that first kind of composition that embodiment 5 obtains can stop ethanol to cause liver liver GSH to exhaust effectively and MDA raises, and alleviates hepatic cell fattydegeneration, has prevention ethanol liver damage function.Utilize first kind of composition of second kind of composition, embodiment 1 of embodiment 6, embodiment 5 and the third composition of embodiment 2 to carry out above-mentioned identical test, obtained roughly the same result.
Embodiment 10: the test of the function of the composition strengthening immunity that contains dibydro myricetrin and ampelopsin of the present invention
1. contain of the influence of the composition of dibydro myricetrin and ampelopsin to the Turnover of Mouse Peritoneal Macrophages phagocytic function
1.1 experiment purpose
Observation contains the influence of the composition of dibydro myricetrin and ampelopsin to the mouse phagocytic function.
1.2 be subjected to the reagent thing
Title: the composition that contains dibydro myricetrin and ampelopsin
Solvent: hot distilled water
Consumption: 0.5ml/20g mouse
1.3 control sample
Biphenylylmethylcarbinol; Shanghai balance pharmaceutical factory, the accurate word (1995) of medicine is defended No. 3765-011 in Shanghai
1.4 animal
Mouse, the Kunming kind, male, 19～21g.
Chicken red blood cell: 5% concentration
1.5 method
Kunming mouse, be divided into 5 groups at random, it is control group, the composition 75 that contains dibydro myricetrin and ampelopsin, 150,300mg/kg, Biphenylylmethylcarbinol 150mg/kg, control group gives physiological saline, the equal oral administration of administration group, every day 1 time, totally 6 times, abdominal injection 0.5% lactoalbumin hydrolysate 1.5ml/ simultaneously of last administration, pneumoretroperitoneum was injected 5% chicken red blood cell 0.1ml in 24 hours, the bloodletting of breaking end after 30 minutes, use the normal saline flushing abdominal cavity, collect peritoneal macrophage and hatch half an hour for 37 ℃, centrifugal, the throw out smear, dyeing, counting cells under the oil mirror calculates with following formula, and compares with control group.
1.6 result
The results are shown in Table 19, the basic, normal, high dosage of composition that contains dibydro myricetrin and ampelopsin all can promote Turnover of Mouse Peritoneal Macrophages to engulf chicken red blood cell and increase phagocytic index, and Biphenylylmethylcarbinol 150mg/kg dosage group also can promote the Turnover of Mouse Peritoneal Macrophages phagocytic function.
Table 19. contains the composition of dibydro myricetrin and ampelopsin
To the influence of Turnover of Mouse Peritoneal Macrophages phagocytic function (± SD)
| Group | Dosage mg/kg | Approach | Number of animals (only) | Phagocytic percentage % | Phagocytic index |
| First kind of composition of the embodiment of the invention 5 | 75 | po×6 | 8 | 32.5±3.5 ** | 0.67±0.08 ** |
| 150 | po×6 | 8 | 43.7±3.9 ** | 0.90±0.03 ** | |
| 300 | po×6 | 8 | 43.7±5.2 ** | 0.84±0.08 ** | |
| Biphenylylmethylcarbinol | 150 | po×6 | 8 | 40.1±4.5 ** | 0.88±0.03 ** |
| Control group | Coordinative solvent | 8 | 23.0±5.3 | 0.39±0.12 | |
*Compare with model control group P＜0.01
2. contain of the influence of the composition of dibydro myricetrin and ampelopsin to mice spleen lymphocytes proliferation
2.1 experiment purpose
Observation contains the influence of the composition of dibydro myricetrin and ampelopsin to mice spleen lymphocytes proliferation.
2.2 be subjected to the reagent thing
Title: the composition that contains dibydro myricetrin and ampelopsin
Solvent: hot distilled water
Consumption: 0.5ml/20g mouse
2.3 control sample
Biphenylylmethylcarbinol; Shanghai balance pharmaceutical factory, the accurate word (1995) of medicine is defended No. 3765-011 in Shanghai
2.4 animal
Mouse, C
57BL/6, ♂, 19～21g.
2.5 other materials
Canavalin(e) (ConA): Sigma product, 50 μ g/ml concentration
3H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml
Substratum: RPMI-1640 includes 15% calf serum, mercaptoethanol, Hepes etc.
2.6 method
C
57The BL/6 mouse, be divided into 5 groups at random, be the normal control group, contain composition 75,150,300mg/kg, the Biphenylylmethylcarbinol 150mg/kg of dibydro myricetrin and ampelopsin, every day, oral administration was 1 time, continuous 7 days, after administration finishes, put to death under the animal aseptic condition and get spleen, counting Pi cell, and the adjustment cell concn is 1 * 10
7/ ml, every hole adds cell suspension 100 μ l on 96 porocyte culture plates, ConA 50 μ l, and nutrient solution, each group is all established three multiple pipes, 37 ℃, 5%CO
2Cultivated 48 hours under the condition, add
3H-TdR 0.5 μ ci/ hole continues to cultivate 18 hours, with bull cell harvestor collecting cell, surveys the CPM value on liquid scintillation instrument, and compares with control group, the results are shown in Table 20.
2.7 result
Table 20. contains the composition of dibydro myricetrin and ampelopsin
Influence to mice spleen lymphocytes proliferation
| Group | Dosage mg/kg | Approach | Number of animals (only) | The CPM value (± SD) |
| First kind of composition of the embodiment of the invention 5 | 75 | po×7 | 4 | 11956±2178 ** |
| 150 | po×7 | 4 | 7869±1401 * | |
| 300 | po×7 | 4 | 7947±1321 * | |
| Biphenylylmethylcarbinol | 150 | po×7 | 4 | 7188±1010 ** |
| Control group | N.S. | 4 | 5448±917 | |
*P＜0.05
*Compare with control group P＜0.01
The result shows that the composition that contains dibydro myricetrin and ampelopsin has the effect of tangible promotion mice spleen lymphocytes proliferation, and strong than other two groups with the low dosage effect.
Composition with embodiment 1, embodiment 2 and embodiment 6 is tested, and has obtained roughly the same test-results, illustrates that these compositions all have the function of strengthening immunity.
Embodiment 11: the composition that contains embodiment 2 is as 1000 milliliters of the little jujube wine of the preparation of the fruit wine of additive,
First kind of composition 0.5g that embodiment 5 obtains,
Said components stirs, and obtains former little jujube wine natural colored liquid, and it is the same with the little jujube wine that does not add first kind of composition that embodiment 5 obtains to drink the sense of taste.Obtain identical result with embodiment 1 with 5 composition respectively.
Embodiment 12: contain first kind of composition that embodiment 5 obtains as 1000 milliliters in the preparation Maotai of the liquor of additive,
First kind of composition 10g that embodiment 5 obtains,
Said components stirs, and obtains the liquid of clear, and it is the same with the Maotai that does not add first kind of composition that embodiment 5 obtains to drink the sense of taste.Obtain identical result with embodiment 1 with 2 composition respectively.
Embodiment 13: contains first kind of composition that embodiment 5 obtains and is mixed with 56 degree liquor 1000ml with medical alcohol and water, add first kind of composition 0.5g that embodiment 5 obtains as the preparation of the liquor 1 of additive,
Said components stirs, and obtains the liquid of clear, and it is the same with the Erguotou wine that does not add first kind of composition that embodiment 5 obtains to drink the sense of taste.Obtain identical result with embodiment 1 with 2 composition respectively.
Embodiment 14: contains first kind of composition that embodiment 5 obtains and is mixed with 38 degree liquor 1000ml with medical alcohol and water, add first kind of composition 1g that embodiment 5 obtains as the preparation of the liquor 2 of additive,
Said components stirs, and obtains the liquid of clear, and it is the same with the Erguotou wine that does not add first kind of composition that embodiment 5 obtains to drink the sense of taste.Obtain identical result with embodiment 1 with 2 composition respectively.
Embodiment 15: contains first kind of composition that embodiment 5 obtains and is mixed with 56 degree liquor 1000ml with medical alcohol and water, add first kind of composition 2g that embodiment 5 obtains as the preparation of the liquor 3 of additive,
Said components stirs, and obtains the liquid of clear, and it is the same with the Erguotou wine that does not add first kind of composition that embodiment 5 obtains to drink the sense of taste.Obtain identical result with embodiment 1 with 2 composition respectively.
Claims (12)
1. health promoting wine that is added with the composition that contains dibydro myricetrin and ampelopsin, be characterised in that the dihydromyricetin cellulose content is 20% to 98% weight ratio in the wherein said composition that contains dibydro myricetrin and ampelopsin, ampelopsin content is 2% to 80% weight ratio.
2. the health promoting wine of claim 1 is characterised in that also containing one or more in the wherein said composition that contains dibydro myricetrin and ampelopsin is selected from following composition: arbutin, gallic acid and/or tea-polyphenol.
3. the health promoting wine of claim 2, the content that is characterised in that arbutin in the wherein said composition that contains dibydro myricetrin and ampelopsin is 0-50wt%, the content of gallic acid is 0-50wt%, the content of tea-polyphenol is 0-50wt%, condition is that the content of arbutin, gallic acid, three kinds of compositions of tea-polyphenol is not zero simultaneously.
4. the health promoting wine of claim 3 is characterised in that the wherein said composition that contains dibydro myricetrin and ampelopsin extracts from ampelopsis.
5. the health promoting wine of claim 4 is characterised in that wherein said ampelopsis comprises ampelopsis cantoniensis, a leaf beautiful grape, Ampelopsis grossedentata, Cayratia japonica (Thunb.) Gagnep., radix ampelopsis, Northeastern Caulis seu folium ampelopsis brevipedunculatae or Vitis Amurensis.
6. the health promoting wine of claim 1 is characterised in that the wherein said composition that contains dibydro myricetrin and ampelopsin is mixed in proportion each monomer component to obtain.
7. claim 1 or 2 health promoting wine are characterised in that the composition levels that contains dibydro myricetrin and ampelopsin in the wherein said wine is the 0.1-53% grams per liter.
8. the health promoting wine of claim 7 is characterised in that the wherein said composition levels that contains dibydro myricetrin and ampelopsin is that 2% grams per liter is to the described amount that contains the degree of dissolution saturation of composition in various wine of dibydro myricetrin and ampelopsin.
9. claim 1 or 2 health promoting wine are characterised in that it is a kind of alcohol wine.
10. the composition that contains dibydro myricetrin and ampelopsin is as the purposes of additive in the various wine of preparation, the dihydromyricetin cellulose content is 20% to 98% weight ratio in the wherein said composition, ampelopsin content is 2% to 80% weight ratio, and randomly, add arbutin, gallic acid and/or tea-polyphenol in the said composition.
11. the composition that contains dibydro myricetrin and ampelopsin prevents and/or treats purposes in the health promoting wine of hyperlipidemia in preparation, the dihydromyricetin cellulose content is 20% to 98% weight ratio in the wherein said composition, ampelopsin content is 2% to 80% weight ratio, and randomly, add arbutin, gallic acid and/or tea-polyphenol in the said composition.
12. the composition that contains dibydro myricetrin and ampelopsin prevents and/or treats purposes in the health promoting wine of alcoholic liver injury in preparation, the dihydromyricetin cellulose content is 20% to 98% weight ratio in the wherein said composition, ampelopsin content is 2% to 80% weight ratio, and randomly, add arbutin, gallic acid and/or tea-polyphenol in the said composition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031238882A CN1304548C (en) | 2003-05-30 | 2003-05-30 | Health care wine containing dihydro-myricetin and myricetin composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031238882A CN1304548C (en) | 2003-05-30 | 2003-05-30 | Health care wine containing dihydro-myricetin and myricetin composition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1483801A CN1483801A (en) | 2004-03-24 |
| CN1304548C true CN1304548C (en) | 2007-03-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB031238882A Expired - Fee Related CN1304548C (en) | 2003-05-30 | 2003-05-30 | Health care wine containing dihydro-myricetin and myricetin composition |
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| Country | Link |
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| CN (1) | CN1304548C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100341501C (en) * | 2003-05-30 | 2007-10-10 | 任启生 | Health care food containing dihydromyricetin and myricetin composition |
| BRPI0402260B1 (en) | 2004-06-15 | 2015-02-18 | Botica Com Farmaceutica Ltda | Composition for toiletries, cosmetics and perfumes. |
| CN109007477A (en) * | 2018-08-21 | 2018-12-18 | 王成有 | A kind of alcoholic drink mixed with fruit juice, waxberry powder and red bayberry liquid beverage |
| CN110591872A (en) * | 2019-09-27 | 2019-12-20 | 健士星生物技术研发(上海)有限公司 | Vine tea liver-protecting wine and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000319154A (en) * | 1999-05-06 | 2000-11-21 | Nippon Menaade Keshohin Kk | Phototoxicity inhibitor |
| CN1483327A (en) * | 2003-05-30 | 2004-03-24 | 任启生 | Health care food containing dihydromyricetin and myricetin composition |
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2003
- 2003-05-30 CN CNB031238882A patent/CN1304548C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000319154A (en) * | 1999-05-06 | 2000-11-21 | Nippon Menaade Keshohin Kk | Phototoxicity inhibitor |
| CN1483327A (en) * | 2003-05-30 | 2004-03-24 | 任启生 | Health care food containing dihydromyricetin and myricetin composition |
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| CN1483801A (en) | 2004-03-24 |
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