CN100529043C - Ampelopsin healthcare wine - Google Patents
Ampelopsin healthcare wine Download PDFInfo
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- CN100529043C CN100529043C CNB031238815A CN03123881A CN100529043C CN 100529043 C CN100529043 C CN 100529043C CN B031238815 A CNB031238815 A CN B031238815A CN 03123881 A CN03123881 A CN 03123881A CN 100529043 C CN100529043 C CN 100529043C
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- ampelopsin
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- KJXSIXMJHKAJOD-LSDHHAIUSA-N (+)-dihydromyricetin Chemical compound C1([C@@H]2[C@H](C(C3=C(O)C=C(O)C=C3O2)=O)O)=CC(O)=C(O)C(O)=C1 KJXSIXMJHKAJOD-LSDHHAIUSA-N 0.000 title claims abstract description 180
- KQILIWXGGKGKNX-UHFFFAOYSA-N dihydromyricetin Natural products OC1C(=C(Oc2cc(O)cc(O)c12)c3cc(O)c(O)c(O)c3)O KQILIWXGGKGKNX-UHFFFAOYSA-N 0.000 title claims abstract description 92
- 235000014101 wine Nutrition 0.000 title claims abstract description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000000654 additive Substances 0.000 claims abstract description 16
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- 238000002360 preparation method Methods 0.000 claims abstract description 14
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- 230000036541 health Effects 0.000 claims description 11
- 201000001421 hyperglycemia Diseases 0.000 claims description 11
- 230000001737 promoting effect Effects 0.000 claims description 9
- 208000031226 Hyperlipidaemia Diseases 0.000 claims description 7
- 206010067125 Liver injury Diseases 0.000 claims description 5
- 230000001476 alcoholic effect Effects 0.000 claims description 5
- 231100000753 hepatic injury Toxicity 0.000 claims description 4
- 230000003053 immunization Effects 0.000 claims 1
- 235000015203 fruit juice Nutrition 0.000 abstract description 2
- 229920006395 saturated elastomer Polymers 0.000 abstract 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 42
- 238000012360 testing method Methods 0.000 description 28
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- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 17
- 229960003180 glutathione Drugs 0.000 description 17
- 238000004458 analytical method Methods 0.000 description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 15
- 239000003795 chemical substances by application Substances 0.000 description 14
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- 238000000034 method Methods 0.000 description 14
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 241000700159 Rattus Species 0.000 description 13
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 12
- 108010023302 HDL Cholesterol Proteins 0.000 description 11
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- 150000002632 lipids Chemical class 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 210000005229 liver cell Anatomy 0.000 description 6
- ZKHSPLCIOVILMA-UHFFFAOYSA-N 2-(2-phenylphenyl)ethanol Chemical compound OCCC1=CC=CC=C1C1=CC=CC=C1 ZKHSPLCIOVILMA-UHFFFAOYSA-N 0.000 description 5
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- 235000003969 glutathione Nutrition 0.000 description 4
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- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 230000007306 turnover Effects 0.000 description 4
- KVJHGPAAOUGYJX-UHFFFAOYSA-N 1,1,3,3-tetraethoxypropane Chemical compound CCOC(OCC)CC(OCC)OCC KVJHGPAAOUGYJX-UHFFFAOYSA-N 0.000 description 3
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000287828 Gallus gallus Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241001247821 Ziziphus Species 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
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- 230000002265 prevention Effects 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
- 230000014860 sensory perception of taste Effects 0.000 description 3
- 230000007863 steatosis Effects 0.000 description 3
- 231100000240 steatosis hepatitis Toxicity 0.000 description 3
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 description 2
- YCPXWRQRBFJBPZ-UHFFFAOYSA-N 5-sulfosalicylic acid Chemical compound OC(=O)C1=CC(S(O)(=O)=O)=CC=C1O YCPXWRQRBFJBPZ-UHFFFAOYSA-N 0.000 description 2
- 241000563984 Ampelopsis Species 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 238000012449 Kunming mouse Methods 0.000 description 2
- 108090001030 Lipoproteins Proteins 0.000 description 2
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 235000009388 Parthenocissus quinquefolia Nutrition 0.000 description 2
- TZRXHJWUDPFEEY-UHFFFAOYSA-N Pentaerythritol Tetranitrate Chemical compound [O-][N+](=O)OCC(CO[N+]([O-])=O)(CO[N+]([O-])=O)CO[N+]([O-])=O TZRXHJWUDPFEEY-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- FHNINJWBTRXEBC-UHFFFAOYSA-N Sudan III Chemical compound OC1=CC=C2C=CC=CC2=C1N=NC(C=C1)=CC=C1N=NC1=CC=CC=C1 FHNINJWBTRXEBC-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- HIMXGTXNXJYFGB-UHFFFAOYSA-N alloxan Chemical compound O=C1NC(=O)C(=O)C(=O)N1 HIMXGTXNXJYFGB-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 244000309464 bull Species 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003651 drinking water Substances 0.000 description 2
- 235000020188 drinking water Nutrition 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 230000003118 histopathologic effect Effects 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 229940090044 injection Drugs 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- -1 lipid peroxide Chemical class 0.000 description 2
- 208000019423 liver disease Diseases 0.000 description 2
- 235000020068 maotai Nutrition 0.000 description 2
- 235000012054 meals Nutrition 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 108700022737 rat Fat1 Proteins 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 229940099373 sudan iii Drugs 0.000 description 2
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- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- XYZZKVRWGOWVGO-UHFFFAOYSA-N Glycerol-phosphate Chemical class OP(O)(O)=O.OCC(O)CO XYZZKVRWGOWVGO-UHFFFAOYSA-N 0.000 description 1
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 1
- 241000581650 Ivesia Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 206010068676 Pneumoretroperitoneum Diseases 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 208000005727 Retropneumoperitoneum Diseases 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000004141 Sodium laurylsulphate Substances 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
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- 230000000295 complement effect Effects 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000019990 fruit wine Nutrition 0.000 description 1
- 229940093181 glucose injection Drugs 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 210000005161 hepatic lobe Anatomy 0.000 description 1
- 238000010562 histological examination Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 230000008818 liver damage Effects 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
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 231100000915 pathological change Toxicity 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
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
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- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides one kind of wine with ampeloptin as additive and its preparation process. The wine, including alcohol wine and fruit juice wine, has ampeloptin content of 0.1-53 g/L, preferably from 2 g/L to the saturated dissolubility of ampeloptin. The wine with ampeloptin as additive is prepared by adding ampeloptin directly into wine via stirring.
Description
The invention belongs to field of health care food.
Document Walter Karrerr, Birkhauser Verlag, Bassel undstuttgart (1958), P.652, NO:1640 discloses the structural formula of ampelopsin,
But the activity of ampelopsin is not studied fully.
The applicant is surprisingly found out that ampelopsin has purposes widely in field of health care products.
The inventor finds that ampelopsin has the function of prevention and/or recovery alcoholic liver injury, has liver protecting, the function of enhancing immunity systemic immunity power, also has hypoglycemic and reducing blood lipid, 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.An object of the present invention is to provide a kind of ampelopsin that contains as wine of additive and preparation method thereof.Wine comprises fruit juice wine, blended liquor and fermented wine.Ampelopsin content can be 0.1-53%w/v (percent weight in volume (grams per liter), hereinafter identical) in the wine, and preferably 2%w/v is to the amount of the degree of dissolution saturation of ampelopsin in various wine.Can be directly the ampelopsin of described amount be added in the wine and stirs, make and contain the wine of ampelopsin as additive.Another purpose of wood invention provides ampelopsin as the purposes of additive in the various wine of preparation.
Ampelopsin can be extracted from ampelopsis, also can prepare with above-mentioned document disclosed method.
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: ampelopsin is as the preparation of the fruit wine of additive
1000 milliliters of little jujube wine,
Ampelopsin 0.5g,
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 ampelopsin to drink the sense of taste.
Embodiment 2: ampelopsin is as the preparation of the liquor of additive
Maotai 1000ml,
Ampelopsin 10g,
Said components stirs, and obtains the liquid of clear, and it is the same with the Maotai that does not add ampelopsin to drink the sense of taste.
Embodiment 3: ampelopsin gets the preparation of liquor 1 as additive
Be mixed with 56 degree liquor 1000ml with medical alcohol and water, add ampelopsin 0.5g.
Said components stirs, and obtains the liquid of clear.
Embodiment 4: ampelopsin is as the preparation of the liquor 2 of additive
Be mixed with 38 degree liquor 1000ml with medical alcohol and water, add ampelopsin 1g.
Said components stirs, and obtains the liquid of clear.
Embodiment 5: ampelopsin is as the preparation of the liquor 3 of additive
Be mixed with 52 degree liquor 1000ml with medical alcohol and water, add ampelopsin 2g.
Said components stirs, and obtains the liquid of clear.
Embodiment 6: the function of ampelopsin prevention and/or recovery alcoholic liver injury
1. materials and methods
1.1 test product: the health promoting wine that contains ampelopsin
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 medical ethanol respectively, concentration is respectively 0.5mg/ml, 1.0mg/ml, 2.0mg/ml, see embodiment 3,4 and 5), 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 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 1
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.
Nitrine is received damping fluid
NaN
3 16.25mg
EDTA-N
al 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 2
Mixing, room temperature were placed after 10 minutes, and the 412nm place measures absorbancy.
1.5.3.2 typical curve
Table 3
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 seedling instead and check is added 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 seedling instead and check is added 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 4, 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 4, 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 4, 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 4 ampelopsin is to the influence of mda, reduced glutathion and triglyceride
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 5.
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 5 pair liver fat drips the influence of distribution
Compare with negative control group: ##P＜0.01; Compare with the damage control group: * P＜0.05
3.. conclusion:
Test-results shows that ampelopsin can stop ethanol to cause liver liver GSH exhaustion and MDA, TG to raise effectively, alleviates hepatic cell fattydegeneration, has prevention ethanol liver damage function.
The hypoglycemic test of example 7 ampelopsin
1. materials and methods
1.1. tried thing: the raw material (wherein the content of ampelopsin is 73%) that contains ampelopsin
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 is tried thing 0.25,0.50,1.50g/kg dosage group, is equivalent to 5,10 and 30 times of people's recommended intake respectively, irritates the stomach amount and 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. ampelopsin is to the influence of normal mouse body weight
Table 6 respectively organize normal mouse initial body weight, mid-term body weight, finish body weight
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. ampelopsin is to the influence of normal mouse fasting plasma glucose
Table 7 ampelopsin is to the influence of normal mouse fasting plasma glucose
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. ampelopsin is to the influence of normal mouse postprandial blood sugar
Table 8 ampelopsin is to the influence of normal mouse postprandial blood sugar
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. ampelopsin is to the influence of normal mouse sugar tolerance
Table 9 ampelopsin is to the influence of normal mouse sugar tolerance
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. ampelopsin is to the influence of the hyperglycemia mouse body weight that causes
Table 10 respectively organize the hyperglycemia model mouse initial body weight, mid-term body weight, finish body weight
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. ampelopsin is to the influence of the hyperglycemia mouse fasting plasma glucose that causes
Table 11 ampelopsin is to the influence of hyperglycemia model mouse fasting plasma glucose
Annotate: compare with model control group,
*P＞0.05 (q check)
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. ampelopsin is to the influence of the hyperglycemia mouse postprandial blood sugar that causes
Table 12 ampelopsin is to the influence of hyperglycemia type mouse postprandial blood sugar
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. ampelopsin is to the influence of the hyperglycemia mouse sugar tolerance that causes
Table 13 ampelopsin is to the influence of hyperglycemia model mouse sugar tolerance
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 ampelopsin has the blood sugar regulation effect.
Embodiment 8: the test of ampelopsin reducing blood-fat
Animal is divided into 4 groups at random, normal control group, positive controls, model control group, is tried the thing 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 that model control group serum TC, TG value obviously raise, and the HDL-C value descends, compare with model, the ampelopsin group can obviously reduce serum TC, TG value, and HDL-C is slightly raise, illustrate that ampelopsin can suppress the mouse blood fat rising that high lipoprotein emulsion causes, has effect for reducing fat.
1. material and method
1.1. tried thing: contain the raw material (wherein the content of ampelopsin is 65%) of ampelopsin, be for experiment with the turbid solution that distilled water is mixed with desired concn with 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. the pure mouse material of common basal feed: Wistar.
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 recommend 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 analyzers (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. snake grape grape element is to the influence of hyperlipidemia model the weight of animals: as seen from table .14, 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.
2.2. ampelopsin is to the influence of high blood lipid model rat body weight: as shown in Table 14, compare with control group, ampelopsin does not have obvious influence to rat body weight.
Table 14 ampelopsin is to the influence of high blood lipid model rat body weight (g, x ± s)
*Compare P＜0.01 with model control group
2.3. ampelopsin is to the influence of rat total cholesterol level: as seen from Table 15, 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 15 ampelopsin is to the influence of rat total cholesterol level (mmol/L, x ± s)
*Compare P＜0.05 with model control group,
*Compare P＜0.01 with model control group
2.4. ampelopsin is to the influence of rat content of triglyceride: as seen from Table 16, 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 16 ampelopsin is to the influence of rat content of triglyceride (mmol/L, x ± s)
*Compare P＜0.05 with model control group,
*Compare P＜0.01 with model control group
2.5. ampelopsin is to the influence of rat high density lipoprotein cholesterol content: as seen from Table 17, 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 17 ampelopsin is to the influence of rat high density lipoprotein cholesterol content (mmol/L1 x ± s)
*Compare P＜0.05 with model control group
2.6. ampelopsin 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 18 as seen, middle and high dosage group total cholesterol level fall is respectively 15.8%, 20.2%, basic, normal, high dosage group content of triglyceride fall is respectively 15.9%, 17.3%, 18.0%, and basic, normal, high dosage group high density lipoprotein cholesterol content lift-off value is 4.48,4.98,4.35mg/dl.
Table 18 ampelopsin is to the influence of rat fat level
Annotate: HDL-c lift-off value (mg/dl) is mmol/L * 38.7=mg/dl conversion result
Embodiment 9: the test of ampelopsin strengthening immunity function
1. ampelopsin is to the influence of Turnover of Mouse Peritoneal Macrophages phagocytic function
1.1 experiment purpose
Observe the influence of ampelopsin to the mouse phagocytic function.
1.2 be subjected to the reagent thing
Title: 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, be control group, ampelopsin 75,150,300mg/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 ampelopsin can promote all Turnover of Mouse Peritoneal Macrophages to engulf chicken red blood cell and increase phagocytic index that Biphenylylmethylcarbinol 150mg/kg dosage group also can promote the Turnover of Mouse Peritoneal Macrophages phagocytic function.
Table 19. ampelopsin to the influence of Turnover of Mouse Peritoneal Macrophages phagocytic function (± SD)
*Compare with model control group P＜0.01
2. ampelopsin is to the influence of mice spleen lymphocytes proliferation
2.1 experiment purpose
Observe the influence of ampelopsin to mice spleen lymphocytes proliferation.
2.2 be subjected to the reagent thing
Title: 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 is divided into 5 groups at random, i.e. normal control group, ampelopsin 75,150,300mg/kg, Biphenylylmethylcarbinol 150 mg/kg, every day, oral administration was 1 time, and continuous 7 days, after administration finishes, put to death under the animal aseptic condition and get spleen, the counting splenocyte, 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. ampelopsin is to the influence of mice spleen lymphocytes proliferation
*P＜0.05
*Compare with control group P＜0.01
The result shows that Stem or leaf of Amur Ampelopsis have the effect of tangible promotion mice spleen lymphocytes proliferation, and strong than other two groups with the low dosage effect.
Embodiment 10: acute toxicity test
After getting mouse prerun, alternative is got 20 mouse, and male and female half and half are given the mouse stomach administration with the peak concentration and the maximum volume of ampelopsin, and 7d is observed in 24h administration 3 times continuously, situations such as record mouse crawler behavior, stool and urine and diet.Result: observed 7 days, and do not see that animal had situations such as dystropy, death, body weight normal growth.Calculating its maximum filling stomach amount is 26.0gkg
-1
Claims (7)
1. the health promoting wine of an ampelopsin is characterised in that wherein ampelopsin content is the 0.1-53% grams per liter.
2. the health promoting wine of claim 1 is characterised in that it is a kind of alcohol wine.
3. each the preparation method of health promoting wine of claim 1-2 is characterised in that directly that ampelopsin with described amount is added in the wine to stir, and makes and contains the wine of ampelopsin as additive.
4. ampelopsin prevents and/or treats purposes in the alcoholic liver injury health promoting wine as additive in preparation, is characterised in that wherein ampelopsin content is the 0.1-53% grams per liter.
5. ampelopsin prevents and/or treats purposes in the health promoting wine of hyperglycemia as additive in preparation, is characterised in that wherein ampelopsin content is the 0.1-53% grams per liter.
6. ampelopsin prevents and/or treats purposes in the health promoting wine of hyperlipidemia as additive in preparation, is characterised in that wherein ampelopsin content is the 0.1-53% grams per liter.
7. ampelopsin prevents and/or treats purposes in the health promoting wine that improves immunizing power as additive in preparation, is characterised in that wherein ampelopsin content is the 0.1-53% grams per liter.
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Non-Patent Citations (6)
| Title |
|---|
| 显齿蛇葡萄基本成分研究. 张友胜，杨伟丽，熊皓平.天然产物研究与开发，第13卷第5期. 2001 |
| 显齿蛇葡萄基本成分研究. 张友胜，杨伟丽，熊皓平.天然产物研究与开发，第13卷第5期. 2001 * |
| 瑶族藤茶中总黄酮类成分的含量测定. 何桂霞，何群，裴刚，邓剑奇.湖南中医药导报，第5卷第12期. 1999 |
| 瑶族藤茶中总黄酮类成分的含量测定. 何桂霞，何群，裴刚，邓剑奇.湖南中医药导报，第5卷第12期. 1999 * |
| 藤茶中APS对小鼠淋巴细胞增殖反应的影响. 曾春晖，杨，柯.广西中医学院学报，第5卷第1期. 2002 |
| 藤茶中APS对小鼠淋巴细胞增殖反应的影响. 曾春晖，杨，柯.广西中医学院学报，第5卷第1期. 2002 * |
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