CN105920057A - Application of icariin - Google Patents
Application of icariin Download PDFInfo
- Publication number
- CN105920057A CN105920057A CN201610386886.7A CN201610386886A CN105920057A CN 105920057 A CN105920057 A CN 105920057A CN 201610386886 A CN201610386886 A CN 201610386886A CN 105920057 A CN105920057 A CN 105920057A
- Authority
- CN
- China
- Prior art keywords
- icariin
- pharmaceutical composition
- ginkgetin
- gemfibrozil
- lovastatin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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- TZJALUIVHRYQQB-XFDQAQKOSA-N Icariin Natural products O(C)c1ccc(C2=C(O[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)C(=O)c3c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O4)c(C/C=C(\C)/C)c3O2)cc1 TZJALUIVHRYQQB-XFDQAQKOSA-N 0.000 title claims abstract description 37
- TZJALUIVHRYQQB-XLRXWWTNSA-N icariin Chemical compound C1=CC(OC)=CC=C1C1=C(O[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)C(=O)C2=C(O)C=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-XLRXWWTNSA-N 0.000 title claims abstract description 37
- TZJALUIVHRYQQB-UHFFFAOYSA-N icariine Natural products C1=CC(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(C)O2)O)C(=O)C2=C(O)C=C(OC3C(C(O)C(O)C(CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-UHFFFAOYSA-N 0.000 title claims abstract description 37
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 claims abstract description 8
- HEMJJKBWTPKOJG-UHFFFAOYSA-N Gemfibrozil Chemical compound CC1=CC=C(C)C(OCCCC(C)(C)C(O)=O)=C1 HEMJJKBWTPKOJG-UHFFFAOYSA-N 0.000 claims abstract description 8
- SQGLUEWZRKIEGS-UHFFFAOYSA-N Ginkgetin Natural products C1=CC(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(OC)C(C=3C(=CC=C(C=3)C=3OC4=CC(O)=CC(O)=C4C(=O)C=3)O)=C2O1 SQGLUEWZRKIEGS-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 claims abstract description 8
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000003613 bile acid Substances 0.000 claims abstract description 8
- 239000000945 filler Substances 0.000 claims abstract description 8
- 229960003627 gemfibrozil Drugs 0.000 claims abstract description 8
- AIFCFBUSLAEIBR-UHFFFAOYSA-N ginkgetin Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C(C=1)=CC=C(OC)C=1C1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 AIFCFBUSLAEIBR-UHFFFAOYSA-N 0.000 claims abstract description 8
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 claims abstract description 8
- 229960004844 lovastatin Drugs 0.000 claims abstract description 8
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 claims abstract description 8
- 206010003210 Arteriosclerosis Diseases 0.000 claims abstract description 5
- 208000011775 arteriosclerosis disease Diseases 0.000 claims abstract description 5
- 239000002778 food additive Substances 0.000 claims abstract description 3
- 235000013373 food additive Nutrition 0.000 claims abstract description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 9
- 235000013402 health food Nutrition 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 4
- 239000000203 mixture Substances 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- 210000004509 vascular smooth muscle cell Anatomy 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 7
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- 108050006400 Cyclin Proteins 0.000 description 5
- 102000009339 Proliferating Cell Nuclear Antigen Human genes 0.000 description 5
- 231100000673 dose–response relationship Toxicity 0.000 description 5
- 230000022131 cell cycle Effects 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102000043136 MAP kinase family Human genes 0.000 description 3
- 108091054455 MAP kinase family Proteins 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 102000019149 MAP kinase activity proteins Human genes 0.000 description 2
- 108040008097 MAP kinase activity proteins Proteins 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
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- -1 flavone compound Chemical class 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000026731 phosphorylation Effects 0.000 description 2
- 238000006366 phosphorylation reaction Methods 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
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- 238000000926 separation method Methods 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 2
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- 238000001262 western blot Methods 0.000 description 2
- IAVUBSCVWHLRGE-UXEKTNMQSA-N (6e)-2,5-dihydroxy-6-[(e)-1-hydroxy-3-(4-hydroxyphenyl)prop-2-enylidene]-2,4-bis[(2s,3r,4r,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]cyclohex-4-ene-1,3-dione Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C(C(C(O)([C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C1=O)=O)=C(O)\C1=C(/O)\C=C\C1=CC=C(O)C=C1 IAVUBSCVWHLRGE-UXEKTNMQSA-N 0.000 description 1
- 241000133570 Berberidaceae Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241001362421 Epimedium brevicornu Species 0.000 description 1
- 206010063560 Excessive granulation tissue Diseases 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000003143 atherosclerotic effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 210000001126 granulation tissue Anatomy 0.000 description 1
- ZZMASNSDVDSYKO-UHFFFAOYSA-N hydroxysafflor yellow A Natural products OCC1OC(C(O)C(O)C1O)C2=C(O)C(O)(C3OC(CO)C(O)C(O)C3O)C(=O)C(=C2O)C(=O)C=Cc4ccc(O)cc4 ZZMASNSDVDSYKO-UHFFFAOYSA-N 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000006101 laboratory sample Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 210000002189 macula lutea Anatomy 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 108010071584 oxidized low density lipoprotein Proteins 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
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- 239000000523 sample Substances 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Emergency Medicine (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of icariin. The icariin can be used for preparing healthcare foods or food additives. A medicine composition for preventing arteriosclerosis by applying the icariin is prepared from the following ingredients by mass percent: 5-15% of icariin, 1-12% of ginkgetin, 1-10% of lovastatin, 5-10% of gemfibrozil, 5-10% of bile acid and 43-83% of a filler.
Description
Technical field
The present invention relates to natural medicine field, be specifically related to the purposes of a kind of icariin.
Background technology
Icariin (Icariin) is the active component extracted from Berberidaceae plant Herba Epimedii (Epimedium brevicornum Maxim) herb, belongs to flavone compound.Icariin has antiinflammatory, antitumor, immunomodulating and antioxidation.
OxLDL ELISA (ox-LDL) can damage by induction of vascular smooth muscle cell (VSMC), and impaired VSMC is migrated, breeds, synthesizes and secretes a large amount of extracellular matrix by middle film under inner membrance, PCNA is i.e. one of important symbol thing of VSMC propagation, and the height of its expression can reflect the proliferation activity of cell.
Mitogen activated protein kinase (MAPK) be intracellular promote propagation and transmission stress the Key kinases of signal, this family includes that the signal path that the subtribes such as ERK, JNK, p38 are relevant to cell proliferation and differentiation is in close relations, this signal transduction pathway, when non-irriate, remains static；When being stimulated by somatomedin, under the effect of upstream kinases, cascade phosphorylation is occurred to activate, MAPK enters nucleus after being activated, make some transcription factor phosphorylations, change the expression status of cytogene, participate in cell growth, break up, the regulation and control of the various physiological processes such as propagation, play an important role in atherosclerotic onset and evolution.
Therefore, regulatory mechanism is bred based on known VSMC, find one of pharmaceutical intervention pathological process Critical policies having become as current prevention and cure of cardiovascular disease related to this, such as " S-A Hydroxysafflor yellow A is expressed and MEK-ERK1/2 signal path suppression proliferation of vascular smooth muscle cells by affecting PCNA " (" Chinese Pharmacological Bulletin " dripped for 7 phases in 2015).
Summary of the invention
The present invention solves that above-mentioned technical problem provides the purposes of a kind of icariin of a kind of icariin.
The present invention solves that the scheme that above-mentioned technical problem is taked is: the purposes of a kind of icariin, it is characterised in that:
This icariin can be used for preparing health food or food additive.
A kind of pharmaceutical composition preventing arteriosclerosis applying icariin, this pharmaceutical composition is made up of following component and mass percent thereof: icariin 5 ~ 15%, ginkgetin 1 ~ 12%, lovastatin 1 ~ 10, gemfibrozil 5 ~ 10%, bile acid 5 ~ 10%, filler 43 ~ 83%.
As preferably, this pharmaceutical composition is made up of following component and mass percent thereof: icariin 13%, ginkgetin 10%, lovastatin 7%, gemfibrozil 9%, bile acid 10%, filler 70%.
Or, this pharmaceutical composition is made up of following component and mass percent thereof: icariin 10%, ginkgetin 7%, lovastatin 10%, gemfibrozil 8%, bile acid 10%, filler 75%.
Accompanying drawing illustrates:
Fig. 1 icariin can dose-dependent inhibition ox-LDL induction VSMC propagation
Fig. 2 MTT experiment result
Fig. 3 the cell cycle experimental result
Fig. 4 icariin can the PCNA protein expression level of VSMC of reduction ox-LDL proliferative induction of dose dependent
Fig. 5 icariin can the ERK1/2 protein phosphorylation of VSMC of reduction ox-LDL proliferative induction of dose dependent.
Detailed description of the invention
It is described in further detail below in conjunction with drawings and Examples:
Embodiment 1:MTT is tested
Experimental technique:
VSMC during being in logarithmic growth is inoculated in 96 orifice plates, and giving ox-LDL (50 μ g/mL) after 24h stimulates, and gives the icariin (0,10,20,40 μM) of various dose after stimulating 24h.Every hole adds after 24h, 48h 20 μ L (5mg/mL) MTT, puts in incubator, 37 ° of C of temperature, hatch 4h, discard MTT solution, add DMSO (150 μ L/ hole) dissolving crystallized, take out after 15min and be placed in microplate reader, at 490nm wavelength, measure its absorbance (OD value).Administration group cell survival rate below equation calculates: survival rate=(process group OD value/matched group OD value) * 100%, result such as Fig. 1,2.
Embodiment 2:Westernblot is tested
Experimental technique:
VSMC during being in logarithmic growth is inoculated in 6 orifice plates, and giving ox-LDL (50 μ g/mL) after 24h stimulates, and gives the icariin (0 of various dose after 24h, 10,20,40 μMs), put in incubator, 37 ° of C of temperature, hatch 24h, collect cell, make westblot laboratory sample, after BCA test kit protein quantification, add 5 × after sample-loading buffer boils 5 min, SDS-PAGE can be carried out.Prepare 10% SDS-PAGE separation gel, insert comb, every hole adds Denatured protein 20 μ g, Marker well adds the Marker of 10 μ l.Switching on power, voltage stabilizing 120 V, 45 min carry out electrophoresis, and the bromophenol blue swimming in protein sample, to when separation gel lower edge 1 cm, stops electrophoresis.The transfering buffering liquid of pre-cooling, constant current 180 mA transferring film 2 h is added in making sandwich, transferring film groove.Take out pvdf membrane and close 2～3 h in room temperature.The anti-diluents of albumen one such as ERK 1/2, p-ERK 1/2, PCNA, with 4 DEG C of shaking table overnight incubation of pvdf membrane.Washing liquid is rinsed 3 times, each 10 min, and Radix Cochleariae officinalis enzyme labelling goat antirabbit two anti-(1:5000) dilution, with pvdf membrane shaking table 2 h under room temperature.Washing liquid is rinsed 3 times, and each 10 min operate according to ECL test kit description, dropped in equably on pvdf membrane by luminescent solution, react 1 min, and film is finally put into gel imaging system detection, result such as Fig. 4,5..
Embodiment 3: cell cycle is tested
Experimental technique:
VSMC during being in logarithmic growth is inoculated in 6 orifice plates, and giving ox-LDL (50 μ g/mL) after 24h stimulates, and gives the icariin (0 of various dose after 24h, 10,20,40 μMs), put in incubator, 37 ° of C of temperature, hatch 24h, in collection cell to the pipe of 15ml, centrifugal, 10mLPBS washes cell, sedimentation cell is the most resuspended, totally 2 times, and 70% ice ethanol is fixed, after adding PCB solution room temperature Hypotonic treatment, the centrifugal supernatant that goes, after addition RNaseA digestion, adds PI, flow cytometer detection cell cycle, result such as Fig. 1,3.
Wherein, MTT dyeing and passage cell cycle result show, the VSMC propagation that ox-LDL is caused by icariin has significant inhibitory action.Westernblot experimental result confirms, the PCNA protein expression level of the VSMC that icariin can be bred after stimulating with the reductions ox-LDL of dose dependent, and can the reduction ERK1/2 protein phosphorylation of dose dependent.
Embodiment 4
A kind of pharmaceutical composition preventing arteriosclerosis applying icariin, weighs icariin 52g, ginkgetin 35g, lovastatin 28g, gemfibrozil 38g, bile acid 37g, filler 110g respectively, and mix homogeneously makes medicine 1000 parts, every part of 0.3g.
Contrast experiment
Experimental technique:
Take the rat of two arteriosclerosis, using icariin (matched group) and icariin compositions (experimental group) to carry out medicine feed respectively, icariin compositions is made up of following component and mass percent thereof: icariin 13%, ginkgetin 10%, lovastatin 7%, gemfibrozil 9%, bile acid 10%, filler 70%.And at a temperature of 37 DEG C, hatch 24h, found by dissection, result such as table 1 below:
Table 1 contrast experiment
| Stratiform collagen | Macula lutea | Granulation tissue and fibrosis | Blood vessel wall | |
| Experimental group | May be seen indistinctly | Nothing | May be seen indistinctly | Soft |
| Matched group | Clearly visible | May be seen indistinctly | Clearly visible | Hardening |
Although the present invention is open as above with embodiment; but it is not limited to protection scope of the present invention; any those of ordinary skill in the art, in the change done without departing from the spirit and scope of the invention and retouching, all should belong to the protection domain of claims of the present invention.
Claims (3)
1. the purposes of an icariin, it is characterised in that: this icariin can be used for preparing health food or food additive.
2. the pharmaceutical composition preventing arteriosclerosis applying icariin, it is characterised in that: this pharmaceutical composition is made up of following component and mass percent thereof: icariin 5 ~ 15%, ginkgetin 1 ~ 12%, lovastatin 1 ~ 10, gemfibrozil 5 ~ 10%, bile acid 5 ~ 10%, filler 43 ~ 83%.
Pharmaceutical composition the most according to claim 1, it is characterised in that: this pharmaceutical composition is made up of following component and mass percent thereof: icariin 13%, ginkgetin 10%, lovastatin 7%, gemfibrozil 9%, bile acid 10%, filler 70%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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