CN106994131B - A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy - Google Patents
A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy Download PDFInfo
- Publication number
- CN106994131B CN106994131B CN201710265740.1A CN201710265740A CN106994131B CN 106994131 B CN106994131 B CN 106994131B CN 201710265740 A CN201710265740 A CN 201710265740A CN 106994131 B CN106994131 B CN 106994131B
- Authority
- CN
- China
- Prior art keywords
- paqg
- fat
- compound
- ebp
- differentiation
- 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.)
- Expired - Fee Related
Links
- 150000002632 lipids Chemical class 0.000 title claims abstract description 26
- 150000001875 compounds Chemical class 0.000 title claims description 51
- 210000001789 adipocyte Anatomy 0.000 claims abstract description 35
- 230000004069 differentiation Effects 0.000 claims abstract description 33
- 239000003814 drug Substances 0.000 claims abstract description 30
- 230000014509 gene expression Effects 0.000 claims abstract description 23
- 238000002360 preparation method Methods 0.000 claims abstract description 23
- 229940079593 drug Drugs 0.000 claims abstract description 21
- 238000009825 accumulation Methods 0.000 claims abstract description 13
- 230000003647 oxidation Effects 0.000 claims abstract description 12
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 12
- 102000011690 Adiponectin Human genes 0.000 claims abstract description 11
- 108010076365 Adiponectin Proteins 0.000 claims abstract description 11
- 102100034808 CCAAT/enhancer-binding protein alpha Human genes 0.000 claims abstract description 10
- 101710168309 CCAAT/enhancer-binding protein alpha Proteins 0.000 claims abstract description 10
- 102100034798 CCAAT/enhancer-binding protein beta Human genes 0.000 claims abstract description 10
- 101710134031 CCAAT/enhancer-binding protein beta Proteins 0.000 claims abstract description 10
- 102100030431 Fatty acid-binding protein, adipocyte Human genes 0.000 claims abstract description 10
- 101001062864 Homo sapiens Fatty acid-binding protein, adipocyte Proteins 0.000 claims abstract description 10
- 102000000536 PPAR gamma Human genes 0.000 claims abstract description 10
- 108010016731 PPAR gamma Proteins 0.000 claims abstract description 10
- 102000040945 Transcription factor Human genes 0.000 claims abstract description 10
- 108091023040 Transcription factor Proteins 0.000 claims abstract description 10
- 230000004136 fatty acid synthesis Effects 0.000 claims abstract description 9
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 9
- 208000030159 metabolic disease Diseases 0.000 claims abstract description 9
- 230000002596 correlated effect Effects 0.000 claims abstract description 8
- 230000001105 regulatory effect Effects 0.000 claims abstract description 8
- 230000002265 prevention Effects 0.000 claims abstract description 3
- 239000008280 blood Substances 0.000 claims description 6
- 210000004369 blood Anatomy 0.000 claims description 6
- 230000009471 action Effects 0.000 claims description 5
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 claims description 5
- 230000003579 anti-obesity Effects 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims 2
- 101000783681 Homo sapiens 5'-AMP-activated protein kinase catalytic subunit alpha-2 Proteins 0.000 abstract description 21
- 210000004027 cell Anatomy 0.000 abstract description 15
- 235000013305 food Nutrition 0.000 abstract description 9
- 230000036541 health Effects 0.000 abstract description 7
- -1 oleanolic acid compound Chemical class 0.000 abstract description 6
- 241000218978 Populus deltoides Species 0.000 abstract description 5
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 abstract description 3
- 208000008589 Obesity Diseases 0.000 abstract description 3
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 abstract description 3
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 abstract description 3
- 235000020824 obesity Nutrition 0.000 abstract description 3
- 229940100243 oleanolic acid Drugs 0.000 abstract description 3
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 abstract description 3
- 230000004913 activation Effects 0.000 abstract description 2
- 230000001276 controlling effect Effects 0.000 abstract description 2
- 208000016097 disease of metabolism Diseases 0.000 abstract description 2
- 102100036009 5'-AMP-activated protein kinase catalytic subunit alpha-2 Human genes 0.000 abstract 1
- SKKCWALCJXDUKO-UHFFFAOYSA-N Pyrocincholic acid Natural products CC1(C)CCC2(CCC3=C(CCC4C5(C)CCC(O)C(C)(C)C5CCC34C)C2C1)C(=O)O SKKCWALCJXDUKO-UHFFFAOYSA-N 0.000 abstract 1
- 235000013376 functional food Nutrition 0.000 abstract 1
- CESLMJOIHBNCGO-UHFFFAOYSA-N pyrocincholic acid 3beta-O-beta-D-quinovopyranosyl-28-O-beta-D-glucopyranoside Natural products OC1C(O)C(O)C(C)OC1OC1C(C)(C)C(CCC2(C)C3=C(C4CC(C)(C)CCC4(CC3)C(=O)OC3C(C(O)C(O)C(CO)O3)O)CCC32)C3(C)CC1 CESLMJOIHBNCGO-UHFFFAOYSA-N 0.000 description 69
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 39
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 36
- 238000002474 experimental method Methods 0.000 description 19
- 102100038074 5'-AMP-activated protein kinase subunit beta-1 Human genes 0.000 description 16
- 239000000523 sample Substances 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 12
- 230000006698 induction Effects 0.000 description 10
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 206010012601 diabetes mellitus Diseases 0.000 description 7
- 239000002775 capsule Substances 0.000 description 6
- 238000001962 electrophoresis Methods 0.000 description 6
- 235000021588 free fatty acids Nutrition 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 235000019359 magnesium stearate Nutrition 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- 101000833892 Homo sapiens Peroxisomal acyl-coenzyme A oxidase 1 Proteins 0.000 description 4
- 101100041816 Homo sapiens SCD gene Proteins 0.000 description 4
- 101000734339 Homo sapiens [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 4, mitochondrial Proteins 0.000 description 4
- 102100026798 Peroxisomal acyl-coenzyme A oxidase 1 Human genes 0.000 description 4
- 101150097713 SCD1 gene Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 102100028897 Stearoyl-CoA desaturase Human genes 0.000 description 4
- 102100034825 [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 4, mitochondrial Human genes 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000010832 independent-sample T-test Methods 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000013139 quantization Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000003757 reverse transcription PCR Methods 0.000 description 4
- 238000007873 sieving Methods 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 150000003626 triacylglycerols Chemical class 0.000 description 4
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 3
- 239000012752 auxiliary agent Substances 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 101710153593 Albumin A Proteins 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 238000000889 atomisation Methods 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 240000002717 Metadina trichotoma Species 0.000 description 1
- 241001529246 Platymiscium Species 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 241001107098 Rubiaceae Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 238000005453 pelletization Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 235000011497 sour milk drink Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000001550 time effect Effects 0.000 description 1
- 150000008130 triterpenoid saponins Chemical class 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
Classifications
-
- 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
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
A kind of 3 β-O- β-D-quinovopyranosyl-28-O- β-D-glucopyranoside (PAQG) of drop oleanolic acid compound Pyrocincholic Acid being extracted from yellow Populus deltoides new function and its application in the drugs, food and health care product that preparation improves fat and fat correlated metabolism diseases for regulating and controlling lipid metaboli in 3T3-L1 cell are provided.PAQG inhibits the differentiation of 3T3-L1 PECTORAL LIMB SKELETON by activation AMPK, lowers the expression of differentiation associated transcription factor and albumen (PPAR γ, C/EBP β, C/EBP α and FABP4) and the release of adiponectin.Meanwhile also by inhibiting fatty acid synthesis and lipid oxidation being promoted to inhibit the accumulation of fat in 3T3-L1 mature fat cell.Under safely and effectively dosage, PAQG improves obesity in terms of adipocyte count and fat content size two, can be used for preparing drug, health care product and the functional food of prevention and treatment obesity and fat relevant metabolic disease.
Description
Technical field:
The invention belongs to technical field of pharmaceuticals, and in particular to a kind of drop oleanolic acid compound Pyrocincholic Acid
3 β-O- β-D-quinovopyranosyl-28-O- β-D-glucopyranoside (PAQG) regulate and control rouge in 3T3-L1 cell
The new function of metabolism and its in the drugs, food and health care product that preparation improves fat and fat correlated metabolism diseases
Using.
Technical background:
Diabetes seriously jeopardize the health of the mankind as a kind of chronic metabolic disease, its complication.Especially
Be the diabetes B closely related with human life style research it is also more and more.Studies have shown that 50% obese people is all
With diabetes B, and the fat illness rate that can make diabetes B increases by 3 times.So the fat research with diabetes
Also it is increasingly concerned.
Obesity is clearly defined as " excessive or abnormal accumulation of fat in vivo " by the World Health Organization (WHO), and causes body
From the aspect of the content two that the reason of interior accumulation of fat can be fatty from adipocyte count and fat cell.To accumulation of fat
Research also become one treatment diabetes novel targets.Although improving fat drug there is various on the market,
Mostly since drug effect is low, side effect is big and unsalable, so, study a kind of medicine for safely and effectively improving fat treatment diabetes
Object or compound are still extremely necessary.
PAQG is a kind of novel drop oleanolic acid type saponin small molecule, is extracted from yellow Populus deltoides.Yellow Populus deltoides is Rubiaceae
Single platymiscium in yellow Populus deltoides category, is grown on southwest China, Vietnam, the ground such as India extensively.Currently, this kind of compounds are only
More than 20 kinds are isolated, PAQG is exactly one such.Moreover, the activity research of these compounds is also fewer.It is of the invention first
It is secondary to have found PAQG in 3T3-L1 cell to the regulating and controlling effect of lipid metaboli.
Summary of the invention:
The purpose of the present invention is to provide a kind of with the compound for significantly inhibiting 3T3-L1 cell differentiation and accumulation of fat
PAQG and the compound are in terms of preparation improves the drug, food and health care product of fat and fat correlated metabolism diseases
Using.
In order to realize above-mentioned purpose of the invention, the present invention provides the following technical solutions:
Application of the compound PAQG in the inhibitor that 3T3-L1 PECTORAL LIMB SKELETON is broken up in preparation,
Compound PAQG is preparing the application in the regulator to mature fat cell fat metabolism.
Compound PAQG is preparing the application in slimming medicine.
Application of the compound PAQG in the drug that preparation adjusts blood lipid.
Application in slimming medicine is being prepared and prepared by the compound PAQG according to the compound PAQG
The application in the drug of blood lipid is adjusted, wherein PAQG is to lower differentiation phase by significantly inhibiting the differentiation of 3T3-L1 PECTORAL LIMB SKELETON
The expression of transcription factor and albumen PPAR γ, C/EBP β, C/EBP α and FABP4 and the release of adiponectin are closed, and passes through inhibition
Fatty acid synthesizes and promotes lipid oxidation to inhibit the accumulation of fat in fat cell, reaches weight-reducing and regulating blood lipid action.
The present invention additionally provides compound PAQG simultaneously improves the fat correlated metabolism of fat and treatment or prevention in preparation
Application in the drug of disease.
Improve medicine that is fat and treating or preventing fat correlated metabolism diseases in preparation according to the compound PAQG
Application in object, wherein PAQG be by significantly inhibit 3T3-L1 PECTORAL LIMB SKELETON differentiation, lower differentiation associated transcription factor and
The expression of albumen PPAR γ, C/EBP β, C/EBP α and FABP4 and the release of adiponectin, and by inhibit fatty acid synthesis and
Promote lipid oxidation to inhibit the accumulation of fat in fat cell, reaches antiobesity action.
The present invention, which still further provides compound PAQG, improves the food of fat and fat correlated metabolism diseases in preparation
And the application in health care product.
By a large amount of multiple independent repeated trials, have found PAQG in 3T3-L1 cell to the regulation of lipid metaboli for the first time
Effect.Present invention discover that PAQG can significantly inhibit the differentiation of 3T3-L1 PECTORAL LIMB SKELETON, differentiation associated transcription factor and egg are lowered
The expression of white (PPAR γ, C/EBP β, C/EBP α and FABP4) and the release of adiponectin.Meanwhile by inhibiting fatty acid synthesis
Inhibit the accumulation of fat in fat cell with lipid oxidation is promoted.Its effect for adjusting lipid metaboli may be with AMPK signal path
It activates related.Under safely and effectively dosage, PAQG improves in terms of adipocyte count and fat content size two
It is fat.
Detailed description of the invention
Fig. 1 oil red O stain.The induction of 3T3-L1 PECTORAL LIMB SKELETON differentiation 6 days.The PAQG of experimental group addition various concentration；
Compound is not added in con group, is substituted with 0.1%DMSO；Undifferentiated group does not add induction agent as blank control.
Experiment is independent in triplicate.
The induction of Fig. 2 .3T3-L1 PECTORAL LIMB SKELETON differentiation 6 days, experimental group added the PAQG of various concentration, being not added of control group
Object is closed, is substituted with 0.1%DMSO.(A) break up associated transcription factor (PPAR γ, C/EBP β and C/EBP α) and albumen FABP4
Electrophoresis result；(B) the adiponectin emission levels in culture medium.Experiment is independent in triplicate, and comparison among groups use independent sample t
It examines, error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
The fat cell that Fig. 3 differentiation is completed, is handled 48 hours, 0.1%DMSO is compared with various concentration PAQG.(A),
Upper figure, medication；The following figure, after administration 48 hours, fat drips content in fat cell；(B), under normal circumstances, 3T3-L1 fat
The time-concentration curve of triglycerides in cell；(C), after drug treatment, in 3T3-L1 fat cell triglycerides when
M- concentration curve；(D), after drug treatment, the time-concentration curve of the free fatty acid in 3T3-L1 fat cell；(E), it gives
After medicine processing, the time-concentration curve of the glycerol in 3T3-L1 fat cell；(F), glycerol release in 3T3-L1 fat cell
Time-concentration curve.Experiment is independent in triplicate, and comparison among groups use independent samples t test, and error line is standard error.With it is right
Compare according to group, P < 0.01 * P < 0.05, * *.
Fig. 4 (A), 3T3-L1 PECTORAL LIMB SKELETON break up in the inductive differentiation medium containing various concentration PAQG, and 6 days
Afterwards, the expression of the albumin A MPK and p-AMPK that are detected with polyacrylamide gel electrophoresis；(B), to the amount of (A) experimental result
Change；(C), 20 μM of PAQG, the DMSO of control group addition 0.1% is added in the overall process of 3T3-L1 PECTORAL LIMB SKELETON induction differentiation,
Then the expression of collection albumen sample detection AMPK and p-AMPK is checked and accepted in different times；(D), to (C) experimental result
Quantization (arrow indicates that compound PAQG starts drug treatment).Experiment is independent in triplicate, and comparison among groups are examined using independent sample t
It tests, error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
Fig. 5 by the PAQG of the 3T3-L1 fat cell various concentration broken up be incubated for 48 hours (collect protein sample and
RT-PCR sample).(A), the electrophoresis result of p-AMPK and AMPK；(B), to the quantization of (A) protein electrophoresis result；(C), RT-PCR
As a result: by the expression of the AMPK gene (SREBP1c, ACC α, SCD1, FAS) relevant to fatty acid synthesis regulated and controled；(D), RT-
PCR result: by the expression of the AMPK gene (PDK4, CPT1a, ACOX1) relevant to lipid oxidation regulated and controled.Experiment is independent to repeat three
Secondary, comparison among groups use independent samples t test, and error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
Fig. 6 is in 3T3-L1 cell, the cytotoxicity experiment of compound PAQG.Experiment is independent in triplicate.
Specific embodiment:
With reference to the accompanying drawing, essentiality content of the invention is further illustrated with the embodiment of the present invention, but not with
This limits the present invention.
Embodiment 1:
The compounds of this invention PAQG is extracted from yellow Populus deltoides shown in structural formula such as following formula (I), by the double versions in Chinese Academy of Sciences west
Receive tropical plants garden offer, preparation method please refers to documented method in following document.
ZHANG Y,TAN N,HUANG H,et al.Triterpenoid Saponins from Metadina
trichotoma[J].Zeitschrift für Naturforschung B,2007,62(5):745‐748.
1, PAQG inhibits the differentiation of 3T3-L1 PECTORAL LIMB SKELETON:
3T3-L1 PECTORAL LIMB SKELETON is two days hungry, with induction agent induction differentiation three days, then with maintenance culture medium culture one
It, replacing fresh culture later and normally cultivating is to be divided into fat cell.According to the experimental method, in entire atomization
It is handled with the PAQG of various concentration (0,2.5 μM, 5 μM, 10 μM, 20 μM), control group is substituted with 0.1% DMSO.It is complete to cell
After full differentiation is good, with the fat content in the method detection fat cell of oil red O stain.As shown in Figure 1, pair with normal differentiation
It is compared according to a group cell, the differentiation of administration group fat cell is obviously inhibited, and concentration gradient dependence is presented, and 20 μM
PAQG is up to 90% to the suppression level of Adipocyte Differentiation, does not observe fat drips substantially in cell at this time.
Fig. 1 oil red O stain.The induction of 3T3-L1 PECTORAL LIMB SKELETON differentiation 6 days.The PAQG of experimental group addition various concentration；
Compound is not added in con group, is substituted with 0.1%DMSO；Undifferentiated group does not add induction agent as blank control.
Experiment is independent in triplicate.
The induction of Fig. 2 .3T3-L1 PECTORAL LIMB SKELETON differentiation 6 days, experimental group added the PAQG of various concentration, being not added of control group
Object is closed, is substituted with 0.1%DMSO.(A) break up associated transcription factor (PPAR γ, C/EBP β and C/EBP α) and albumen FABP4
Electrophoresis result；(B) the adiponectin emission levels in culture medium.Experiment is independent in triplicate, and comparison among groups use independent sample t
It examines, error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
Meanwhile the present invention collects the above protein sample tested, and is detected with agarose gel electrophoresis method, discovery differentiation is related
The expression of transcription factor and albumen (PPAR γ, C/EBP β, C/EBP α and FABP4) is remarkably decreased (Fig. 2A), also, uses adiponectin
Kit detects that the release of adiponectin in culture medium also reduces as shown in Figure 2 B.Result in terms of three above proves PAQG
Significantly inhibit the differentiation of 3T3-L1 PECTORAL LIMB SKELETON.
2, PAQG inhibits the accumulation of fat in mature fat cell
After 3T3-L1 PECTORAL LIMB SKELETON normal differentiation, handled 48 hours with 10 μM and 20 μM of PAQG respectively, control group
For 0.1%DMSO (such as on Fig. 3 A).Observation result is as shown in Figure 3A under the microscope, and compared with cellular control unit, experimental group is thin
Rouge content in born of the same parents significantly reduces.Meanwhile we dissolve with dehydrated alcohol and collect the rouge in cell, and detect wherein glycerol three
The content of ester (TG).As shown in Figure 3B, consistent with microscopic examination result, TG content significant decrease intracellular after PAQG processing, in concentration ladder
Spend dependence.Cell without doing drug treatment, after normal differentiation, TG intracellular is (Fig. 3 B) constantly accumulated.PAQG causes
TG is reduced in fat cell, and the TG hydrolysis in fat cell may be promoted due to it, generates free fatty acid (FFA) and glycerol
(Glycerol), it is discharged into culture medium.Therefore, we have detected the content of FFA and Glycerol in culture medium with kit, such as
Shown in Fig. 3 D and 3E, FFA the and Glycerol content in culture medium is all dramatically increased (compared with Normal group), and is also in
Concentration gradient dependence.
For the fat content variation in dynamic detection PAQG treatment process, in fat cell.We are in PAQG processing
In 48 hours, the glycerol content (Fig. 3 F) in different time points culture medium is detected, and bent with the time that this depicts glycerol release
Line.The results show that in control group and experimental group, the content of glycerol is all to gradually increase as time went on, but chemical combination
After object processing, the content of glycerol is apparently higher than control group, and is in concentration gradient dependence.Result prompt, the lipid-loweringing of PAQG
Effect is related with TG degradation in fat cell is promoted.
The fat cell that Fig. 3 differentiation is completed, is handled 48 hours, 0.1%DMSO is compared with various concentration PAQG.(A),
Upper figure, medication；The following figure, after administration 48 hours, fat drips content in fat cell；(B), under normal circumstances, 3T3-L1 fat
The time-concentration curve of triglycerides in cell；(C), after drug treatment, in 3T3-L1 fat cell triglycerides when
M- concentration curve；(D), after drug treatment, the time-concentration curve of the free fatty acid in 3T3-L1 fat cell；(E), it gives
After medicine processing, the time-concentration curve of the glycerol in 3T3-L1 fat cell；(F), glycerol release in 3T3-L1 fat cell
Time-concentration curve.Experiment is independent in triplicate, and comparison among groups use independent samples t test, and error line is standard error.With it is right
Compare according to group, P < 0.01 * P < 0.05, * *.
3, PAQG activates AMPK signal path in 3T3-L1 cell
When observing that PAQG inhibits 3T3-L1 PECTORAL LIMB SKELETON differentiating phenomenon, we collect the protein sample, and
With the protein expression of agarose gel electrophoresis method detection AMPK and p-AMPK.As illustrated in figures 4 a and 4b, the phosphorylation level of AMPK
Increase as gradient is presented in the concentration variation of PAQG.In order to further probe into AMPK in 3T3-L1 PECTORAL LIMB SKELETON atomization
The situation of change of phosphorylation, We conducted following experiments.Administration when inducing differentiation since just, control group is with 0.1%
DMSO substitution, in day1, day3, day4, day6, these different times check and accept albumen, detect the expression of AMPK and p-AMPK.
As a result as shown in figures 4 c and 4d, PAQG can sustained activation AMPK during the entire process of 3T3-L1 PECTORAL LIMB SKELETON breaks up
Phosphorylation.
In order to probe into the mechanism of action that PAQG inhibits the accumulation of 3T3-L1 cellular fat, We conducted following experiments: when thin
After born of the same parents break up, cell is handled with 10 μM and 20 μM of PAQG respectively, uses 0.1%DMSO as control.Two days later, a part of
Collect the expression of protein sample detection AMPK and p-AMPK；Another part collect RNA sample detection AMPK signal path downstream with
The relevant gene expression of lipid metaboli.Wherein, AMPK signal path gene relevant to lipid metaboli mainly include fatty acid synthesis and
Two class of lipid oxidation.And gene relevant to fatty acid synthesis mainly has: SREBP1c, ACC α, SCD1, FAS；It is related to lipid oxidation
Gene mainly have: PDK4, CPT1a, ACOX1.As shown in figures 5 a-d, the phosphorylation level of AMPK is with PAQG's for experimental result
Concentration variation is presented gradient and increases.In addition, the expression of gene SREBP1c, ACC α, SCD1, FAS reduce, and PDK4 and CPT1a
Expression increase, the expression of ACOX1 is basically unchanged.Thus illustrate, PAQG may inhibit fat by activating AMPK signal path
Acid synthesizes and promotes lipid oxidation, to play effect for reducing fat.
Fig. 4 .A), 3T3-L1 PECTORAL LIMB SKELETON breaks up in the inductive differentiation medium containing various concentration PAQG, and 6 days
Afterwards, the expression of the albumin A MPK and p-AMPK that are detected with polyacrylamide gel electrophoresis；(B), to the amount of (A) experimental result
Change；(C), 20 μM of PAQG, the DMSO of control group addition 0.1% is added in the overall process of 3T3-L1 PECTORAL LIMB SKELETON induction differentiation,
Then the expression of collection albumen sample detection AMPK and p-AMPK is checked and accepted in different times；(D), to (C) experimental result
Quantization (arrow indicates that compound PAQG starts drug treatment).Experiment is independent in triplicate, and comparison among groups are examined using independent sample t
It tests, error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
Fig. 5 by the PAQG of the 3T3-L1 fat cell various concentration broken up be incubated for 48 hours (collect protein sample and
RT-PCR sample).(A), the electrophoresis result of p-AMPK and AMPK；(B), to the quantization of (A) protein electrophoresis result；(C), RT-PCR
As a result: by the expression of the AMPK gene (SREBP1c, ACC α, SCD1, FAS) relevant to fatty acid synthesis regulated and controled；(D), RT-
PCR result: by the expression of the AMPK gene (PDK4, CPT1a, ACOX1) relevant to lipid oxidation regulated and controled.Experiment is independent to repeat three
Secondary, comparison among groups use independent samples t test, and error line is standard error.Compared with the control group, P < 0.01 * P < 0.05, * *.
4, the cytotoxicity experiment of PAQG
The present invention has detected cytotoxicity experiment (Fig. 6) of the PAQG in mouse 3T3-L1 PECTORAL LIMB SKELETON.The result shows that
For compound PAQG without cytotoxicity within the scope of 0~50 μM, the compound maximum concentration used in this experimentation is 20 μM,
This concentration is in safe range.
Fig. 6 is in 3T3-L1 cell, the cytotoxicity experiment of compound PAQG.Experiment is independent in triplicate.
Embodiment 2
By 1 gained compound PAQG of embodiment, figuration is added in the ratio of compound and excipient weight ratio 1:1 or 1:2
Agent, pelletizing press sheet.
Embodiment 3:
Compound PAQG is made by embodiment 1, routinely capsule is made in capsule preparations method.
Embodiment 4:
Compound PAQG first is made by embodiment 1, then tablet is made as follows.
Tablet: compound PAQG100mg, appropriate starch, appropriate corn pulp, Magnesium Stearate proper quantity.Preparation method: by chemical combination
Object is mixed with auxiliary agent, and sieving uniformly mixes in suitable container, obtained granulating mixture tabletting.
Embodiment 5:
Capsule: compound PAQG100mg, appropriate starch, Magnesium Stearate proper quantity.
Preparation method: compound PAQG is mixed with auxiliary agent, and sieving uniformly mixes in suitable container, what is obtained
Mixture is packed into hard gelatin capsule.
Embodiment 6:
Tablet: embodiment 1 gained compound PAQG10mg, lactose 180mg, starch 55mg, magnesium stearate 5mg；
Preparation method: compound PAQG, newborn sugar and starch are mixed, uniformly moistened with water, the mixture mistake after moistening
It sieves and dries, magnesium stearate is added, then by mixture tabletting, every slice weight 250mg, compounds content 10mg in re-sieving.
Embodiment 7:
Ampulla: embodiment 1 gained compound PAQG2mg, sodium chloride 10mg；
Preparation method: compound PAQG and sodium chloride are dissolved in suitable water for injection, acquired solution are filtered, in nothing
It is fitted into ampoule bottle under the conditions of bacterium.
Embodiment 8:
Capsule: embodiment 1 gained compound PAQG10mg, lactose 187mg, magnesium stearate 3mg；
Preparation method: compound is mixed with auxiliary agent, and sieving uniformly mixes, and obtained mixture is packed into glutoid glue
Capsule, each capsule weight 200mg, active component content 10mg.
Embodiment 9:
The preparation of food:
Preparation method: 1 gained compound PAQG of Example is added in jam with the ratio of 1:7, can increase food
Fragrance, while can also play health-care efficacy.
Embodiment 10:
The preparation of food:
Preparation method: 1 gained compound PAQG of Example is added in honey with the ratio of 1:8, can increase food
Fragrance, while can also play health-care efficacy.
Embodiment 11:
Sour milk beverage formula (W%) containing compound PAQG:
Above-mentioned beverage of the invention is made in the method for routinely making food.
Claims (6)
1. compound PAQG is preparing the application in slimming medicine, shown in the compound PAQG structural formula such as formula (I),
2. application of the compound PAQG in the drug that preparation adjusts blood lipid, the compound PAQG structural formula such as formula (I) institute
Show,
3. compound PAQG according to claim 1 is preparing the application in slimming medicine, it is characterised in that PAQG passes through
Significantly inhibit 3T3-L1 PECTORAL LIMB SKELETON differentiation, lower differentiation associated transcription factor and albumen PPAR γ, C/EBP β, C/EBP α and
The expression of FABP4 and the release of adiponectin, and inhibited in fat cell by inhibiting fatty acid synthesis and promoting lipid oxidation
Accumulation of fat, reach antiobesity action.
4. application of the compound PAQG according to claim 2 in the drug that preparation adjusts blood lipid, it is characterised in that
PAQG breaks up associated transcription factor and albumen PPAR γ, C/EBP β by significantly inhibiting the differentiation of 3T3-L1 PECTORAL LIMB SKELETON, downward,
The expression of C/EBP α and FABP4 and the release of adiponectin, and inhibit rouge by inhibiting fatty acid synthesis and promoting lipid oxidation
Accumulation of fat in fat cell, reaches regulating blood lipid action.
5. compound PAQG improves the application in drug that is fat and treating or preventing fat correlated metabolism diseases, institute in preparation
Shown in the compound PAQG structural formula stated such as formula (I),
6. compound PAQG according to claim 5 improves the fat correlated metabolism disease of fat and treatment or prevention in preparation
Application in the drug of disease, it is characterised in that PAQG is broken up by significantly inhibiting 3T3-L1 PECTORAL LIMB SKELETON, and it is related to lower differentiation
The expression of transcription factor and albumen PPAR γ, C/EBP β, C/EBP α and FABP4 and the release of adiponectin, and by inhibiting rouge
Fat acid synthesizes and promotes lipid oxidation to inhibit the accumulation of fat in fat cell, reaches antiobesity action.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710265740.1A CN106994131B (en) | 2017-04-21 | 2017-04-21 | A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710265740.1A CN106994131B (en) | 2017-04-21 | 2017-04-21 | A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106994131A CN106994131A (en) | 2017-08-01 |
| CN106994131B true CN106994131B (en) | 2019-12-03 |
Family
ID=59434441
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710265740.1A Expired - Fee Related CN106994131B (en) | 2017-04-21 | 2017-04-21 | A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106994131B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110272997B (en) * | 2018-03-14 | 2023-04-28 | 华中科技大学同济医学院附属同济医院 | Application of C/EBP beta gene or protein |
| CN108610409B (en) * | 2018-04-09 | 2021-08-24 | 深圳大学 | Application of ETV5 in preparation of medicine for preventing or treating obesity and related metabolic diseases |
| CN109010800A (en) * | 2018-07-05 | 2018-12-18 | 中国科学院深圳先进技术研究院 | PDGF-BB is inhibiting Adipocyte Differentiation, formation and its application of function |
| CN109722421B (en) * | 2018-12-14 | 2022-04-19 | 南京市妇幼保健院 | Adipocyte endogenous polypeptide and application thereof |
| CN110974821A (en) * | 2019-12-24 | 2020-04-10 | 浙江大学 | Application of Eurycomanone and analogues thereof in preparation of medicines for treating hyperlipidemia and fatty liver or weight-reducing products |
| CN111926071B (en) * | 2020-08-14 | 2022-01-04 | 武汉大学 | Molecular marker related to aging and healthy aging and application of molecular marker in improving healthy aging |
| CN114931577A (en) * | 2022-06-21 | 2022-08-23 | 上海市同仁医院 | Application of indole-3-acrylic acid in inhibiting differentiation of white adipose precursor cells |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105147710A (en) * | 2015-09-11 | 2015-12-16 | 中国科学院西双版纳热带植物园 | Hypoglycemic agent as well as preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286057B (en) * | 2011-07-07 | 2016-08-03 | 北华大学 | Oleanane type triterpene compounds and preparation method thereof and medical application |
-
2017
- 2017-04-21 CN CN201710265740.1A patent/CN106994131B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105147710A (en) * | 2015-09-11 | 2015-12-16 | 中国科学院西双版纳热带植物园 | Hypoglycemic agent as well as preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| Triterpenoid Saponins from Metadina trichotoma;Yu-Mei Zhang等;《Z. Naturforsch》;20070501;第32卷(第5期);第745-748页 * |
| 黄棉木中两个新的三萜类化合物;张玉梅等;《云南植物研究》;20061231;第28卷(第6期);第673-675页 * |
| 齐墩果酸和熊果酸的神经精神药理作用研究进展;张明发等;《药物评价研究》;20151031;第38卷(第5期);第570-576页 * |
| 齐墩果酸和熊果酸调血脂、抗肥胖药理作用研究进展;张明发等;《药物评价研究》;20150228;第38卷(第1期);第90-97页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106994131A (en) | 2017-08-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106994131B (en) | A kind of application adjusting lipid metaboli and fat compound PAQG in pharmacy | |
| WO2015172608A1 (en) | Capsule for assisting in reducing blood fat and preparation method therefor | |
| CN102090630A (en) | Health-care product for enhancing antioxidation of human bodies and preparation method thereof | |
| EL-Tantawi et al. | Impact of Spirulina on Propylthiouracil-Induced Hypothyroidism in Albino Rats, a histological, immunohistochemical and biochemical approach | |
| JP2013071909A (en) | Intracerebral lipid peroxide accumulation inhibitor | |
| EP3275457A1 (en) | Pharmaceutical composition containing silybin | |
| Shuai et al. | Hypolipidemic effects of macadamia oil are related to AMPK activation and oxidative stress relief: In vitro and in vivo studies | |
| CN108926595A (en) | A kind of health care product with protection liver and hypolipemic function | |
| WO2017173611A1 (en) | Pharmaceutical product for treating tumor and combined immune defect, and preparation and application thereof | |
| CN105560308B (en) | Flower of JINHUAKUI is preparing the application in the product for preventing and treating prostatic disorders | |
| JP2019532039A (en) | Composition comprising pollen and / or pistil extract, method of preparation and related uses | |
| US10688146B2 (en) | Curcuma mangga val et. zipp. extract as a treatment to overcome prostate problems | |
| KR20170088478A (en) | A composition for antioxidating, antiinflamatory and antidiabetes comprising extracts of inyoolsan | |
| CN109966283A (en) | Application of the degreasing cinnamon polyphenol extract in preparation prevention and treatment diabetic nephropathy product | |
| CN110327323A (en) | Application of the Cichoric acid in preparation treatment obesity or its complication medicine | |
| Ishaka et al. | Toxicity evaluation, HET-CAM irritation, and anti-irritant potential of rice bran wax policosanol nanoemulsion | |
| CN110302340A (en) | A kind of Chinese medicinal tablet and preparation method thereof with hypolipemic function | |
| KR101750112B1 (en) | Compositions for anti-stress comprising oil mixture | |
| CN103768586B (en) | The medicine of a kind of benefit god's brain tonic and preparation thereof | |
| KR102514847B1 (en) | Composition for preventing or treating cognitive dysfunction or neuroinflammation comprising extracts of centella asiatica, cnidium monnieri, and lycium barbarum linne | |
| KR20120121232A (en) | Preparation method of extract of Picrasma quassioides and use of the extracts | |
| NL2031296B1 (en) | Application of aronia melanocarpa leaf polyphenol extract in preparing antidepressants | |
| CN106265640A (en) | APPA application in preparation treatment medicine for treating diabetic nephropathy | |
| Rusdiana et al. | The anti-obesity properties of Anredera cordifolia leaf extract in rats fed a high-fat diet through inhibition of adipogenesis | |
| CN106822114B (en) | Application of MTCA in preparation of medicines for reducing blood sugar or blood fat |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20191203 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |