CN114478821B - Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof - Google Patents
Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof Download PDFInfo
- Publication number
- CN114478821B CN114478821B CN202210198625.8A CN202210198625A CN114478821B CN 114478821 B CN114478821 B CN 114478821B CN 202210198625 A CN202210198625 A CN 202210198625A CN 114478821 B CN114478821 B CN 114478821B
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- radix platycodi
- mice
- platycodon grandiflorum
- water
- 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.)
- Active
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 62
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 62
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 62
- 241000699670 Mus sp. Species 0.000 title claims abstract description 35
- 235000009200 high fat diet Nutrition 0.000 title claims abstract description 17
- 230000006698 induction Effects 0.000 title claims abstract description 14
- 244000274050 Platycodon grandiflorum Species 0.000 claims abstract description 35
- 235000006753 Platycodon grandiflorum Nutrition 0.000 claims abstract description 35
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000001913 cellulose Substances 0.000 claims abstract description 8
- 229920002678 cellulose Polymers 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims abstract description 8
- 208000008589 Obesity Diseases 0.000 claims abstract description 7
- 229920005654 Sephadex Polymers 0.000 claims abstract description 7
- 239000012507 Sephadex™ Substances 0.000 claims abstract description 7
- 235000020824 obesity Nutrition 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 208000031226 Hyperlipidaemia Diseases 0.000 claims abstract description 5
- 238000009825 accumulation Methods 0.000 claims abstract description 5
- 201000001421 hyperglycemia Diseases 0.000 claims abstract description 5
- 238000005238 degreasing Methods 0.000 claims abstract description 4
- 238000001556 precipitation Methods 0.000 claims abstract description 4
- 238000003809 water extraction Methods 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 45
- 239000000243 solution Substances 0.000 claims description 30
- 229910001868 water Inorganic materials 0.000 claims description 22
- 239000012153 distilled water Substances 0.000 claims description 19
- 238000012856 packing Methods 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 10
- 235000006751 Platycodon Nutrition 0.000 claims description 9
- 241000357613 Platycodon Species 0.000 claims description 9
- 210000005228 liver tissue Anatomy 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 230000007935 neutral effect Effects 0.000 claims description 9
- 229930189914 platycodon Natural products 0.000 claims description 9
- 150000002632 lipids Chemical class 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 239000010419 fine particle Substances 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 5
- 238000011068 loading method Methods 0.000 claims description 5
- 239000012982 microporous membrane Substances 0.000 claims description 5
- 239000008346 aqueous phase Substances 0.000 claims description 4
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 235000005911 diet Nutrition 0.000 claims description 4
- 230000037213 diet Effects 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 201000010063 epididymitis Diseases 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 239000004677 Nylon Substances 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 229920001778 nylon Polymers 0.000 claims description 3
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 210000000918 epididymis Anatomy 0.000 claims 1
- 239000000047 product Substances 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 230000003544 deproteinization Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 150000002772 monosaccharides Chemical class 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 230000037396 body weight Effects 0.000 description 6
- 235000019441 ethanol Nutrition 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 4
- 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 4
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical group O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 3
- 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 3
- 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
- 208000033809 Suppuration Diseases 0.000 description 3
- 230000035508 accumulation Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 230000000062 effect on obesity Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- CBOJBBMQJBVCMW-BTVCFUMJSA-N (2r,3r,4s,5r)-2-amino-3,4,5,6-tetrahydroxyhexanal;hydrochloride Chemical compound Cl.O=C[C@H](N)[C@@H](O)[C@H](O)[C@H](O)CO CBOJBBMQJBVCMW-BTVCFUMJSA-N 0.000 description 1
- 241000132012 Atractylodes Species 0.000 description 1
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 description 1
- 241000208671 Campanulaceae Species 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- LKDRXBCSQODPBY-VRPWFDPXSA-N D-fructopyranose Chemical compound OCC1(O)OC[C@@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-VRPWFDPXSA-N 0.000 description 1
- 241001523681 Dendrobium Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 238000013218 HFD mouse model Methods 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 241000414067 Inonotus obliquus Species 0.000 description 1
- 208000007976 Ketosis Diseases 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 229920001218 Pullulan Polymers 0.000 description 1
- 239000004373 Pullulan Substances 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 206010042674 Swelling Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 150000001323 aldoses Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- PBAYDYUZOSNJGU-UHFFFAOYSA-N chelidonic acid Natural products OC(=O)C1=CC(=O)C=C(C(O)=O)O1 PBAYDYUZOSNJGU-UHFFFAOYSA-N 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002270 exclusion chromatography Methods 0.000 description 1
- 238000002474 experimental method Methods 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
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 229960001911 glucosamine hydrochloride Drugs 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 210000003026 hypopharynx Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002584 ketoses Chemical class 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000006372 lipid accumulation Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 201000003453 lung abscess Diseases 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 235000009048 phenolic acids Nutrition 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 229920001197 polyacetylene Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 235000019423 pullulan Nutrition 0.000 description 1
- 235000019633 pungent taste Nutrition 0.000 description 1
- 150000003214 pyranose derivatives Chemical group 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- 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
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- 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)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Diabetes (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Sustainable Development (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Child & Adolescent Psychology (AREA)
- Epidemiology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a platycodon grandiflorum polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof, and relates to a polysaccharide extracted from platycodon grandiflorum, which specifically comprises Ara, glcN, gal, glc, xyl, man and Fru with a molar ratio of 1.1:0.4:0.4:10.9:2.2:0.9:84.1; the preparation method comprises degreasing, water extraction, alcohol precipitation, deproteinization, cellulose column separation and sephadex column separation, wherein the polysaccharide is named as PGNP, has the characteristic of improving the constitution of mice after high-fat diet induction, and mainly comprises the steps of improving obesity, inhibiting fat accumulation, preventing hyperglycemia, hyperlipidemia and the like.
Description
Technical Field
The invention relates to a polysaccharide, in particular to a platycodon grandiflorum polysaccharide.
Background
Polysaccharides are widely found in nature and are organic chemical polymers formed by the final polymerization of more than 10 aldoses or ketoses linked together by glycosidic bonds. The polysaccharide substances have various important biological activities of inhibiting malignant tumors in organisms, resisting oxidation, regulating immune system dysfunction, reducing blood fat and the like, are rich in resources, and can be extracted and separated from different plants and microorganisms to obtain various active polysaccharides such as bighead atractylodes rhizome polysaccharide, dendrobium polysaccharide, inonotus obliquus sporophore polysaccharide, pullulan polysaccharide and the like, so that the polysaccharide component in the traditional Chinese medicinal materials gradually becomes one of important academic hot spots of the application research of modern Chinese medicine science and development technology in China.
The Chinese medicine radix Platycodi (Platycodon grandiflorum (Jacq.) A.DC) is mainly derived from radix Platycodi of genus Platycodi in family Campanulaceae of China, and the main medicinal part is dry root of radix Platycodi. Bitter and pungent taste, neutral nature, and lung meridian entered; has effects of dispersing lung qi, relieving sore throat, eliminating phlegm, and expelling pus, and can be used for treating chest distress, laryngopharynx swelling and pain, pulmonary abscess, and pus discharge etc. The radix Platycodi contains various chemical components such as radix Platycodi flavonoids, saponins, sterols, fatty acids, polyacetylenes, polysaccharides, phenolic acids and other bioactive compounds. Radix Platycodi has remarkable curative effects in relieving cough, resisting common cold, suppurating, expelling pus, lowering blood pressure, reducing tobacco toxicity, etc. Polysaccharide is one of important bioactive substances in platycodon grandiflorum, has little research on lipid lowering, and needs to be deeply studied.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, and provides platycodon grandiflorum polysaccharide capable of improving constitution of mice after high-fat diet induction and application thereof, so as to solve the technical problem that the platycodon grandiflorum polysaccharide in the prior art has less research on lipid lowering.
The invention is realized by the following technical scheme:
the invention provides platycodon grandiflorum polysaccharide capable of improving constitution of mice after induction of high-fat diet, which specifically comprises Ara, glcN, gal, glc, xyl, man and Fru with a molar ratio of 1.1:0.4:0.4:10.9:2.2:0.9:84.1.
The invention also provides a preparation method of the platycodon grandiflorum polysaccharide, which comprises the following steps:
Pulverizing radix Platycodi decoction pieces to obtain radix Platycodi powder, placing 50g of radix Platycodi powder into 500ml round bottom flask, adding 250ml of absolute ethanol, refluxing in 80deg.C water bath for 1 hr, repeating twice, and drying residues in shade to obtain defatted radix Platycodi powder;
step 2, water extraction
Placing 100g of defatted radix Platycodi powder into a 2L round bottom flask, adding 1500ml of distilled water, heating to 80deg.C, extracting with water for 3 times, 1.5 hr each time, filtering the water extractive solution with 300 mesh nylon gauze each time, vacuum filtering, preserving filtrate, mixing three filtrates, and concentrating under heating to obtain radix Platycodi polysaccharide water extract;
step 3, alcohol precipitation
Adding 95% ethanol into radix Platycodi polysaccharide water extract to reach alcohol concentration of 80%, standing at 4deg.C overnight, centrifuging, and removing upper layer solution to obtain radix Platycodi polysaccharide-protein mixture;
step 4, deproteinizing
Adding distilled water into the platycodon polysaccharide-protein mixture to prepare a solution with the concentration of 1mg/mL, mixing the platycodon polysaccharide-protein mixture solution, chloroform and n-butanol according to the volume ratio of 25:4:1 to obtain a mixed solution, electromagnetically stirring the mixed solution for 30min, centrifuging, and freeze-drying an upper aqueous phase solution for 48h to obtain platycodon polysaccharide;
Soaking 35g of DEAE-52 cellulose in 3500mL of degassed distilled water overnight, pouring out fine particles suspended on the upper layer, and preparing for column packing; dissolving 50mg of radix Platycodi crude polysaccharide in 5mL of distilled water, passing through a microporous filter membrane of 0.22 μm, and preparing for sample loading; slowly dripping a sample on the surface of the column packing, fully attaching the sample to the packing, eluting with distilled water with the volume twice that of the column, collecting a tube for 5min at the flow rate of 0.6 mL/min; collecting sugar solution, mixing, concentrating, and freeze drying to obtain radix Platycodi neutral polysaccharide;
step 6, sephadex column separation
Pouring 15g sephadex G100 into 1500mL of de-aerated deionized water, soaking overnight, pouring out the suspended fine particles on the upper layer, and preparing for column loading; dissolving 30mg of neutral polysaccharide of radix Platycodi with 10mL of water, filtering with 0.22 μm microporous membrane, and loading; slowly dripping sample on the surface of column packing, making it fully adhere to the column packing, collecting one tube with distilled water as eluent for 5min, detecting each tube of eluent by phenol-sulfuric acid method, drawing corresponding elution curve, mixing the eluates, concentrating and freeze-drying to obtain uniform radix Platycodi polysaccharide, named PGNP.
The invention also provides application of the platycodon grandiflorum polysaccharide in improving diet or medicine of mice after high-fat diet induction.
The invention also provides application of the platycodon grandiflorum polysaccharide in diet or medicine for improving obesity-related hyperglycemia and hyperlipidemia.
Compared with the prior art, the invention has the following advantages:
the platycodon grandiflorum polysaccharide capable of improving the constitution of the mice after the induction of the high-fat diet and the application thereof can effectively improve the relevant constitution of the mice after the induction of the high-fat diet, and mainly comprises the effects of improving the obesity, inhibiting fat accumulation, preventing hyperglycemia, hyperlipidemia and the like.
Drawings
FIG. 1 is a protein standard graph obtained in example step 4.2;
FIG. 2 is a graph of the standard sugar content obtained in example step 5;
FIG. 3 is a graph showing elution of neutral polysaccharides from platycodon grandiflorum obtained in example step 6;
FIG. 4 is a graph showing the elution profile of PGNP obtained in example step 7;
FIG. 5 is a liquid chromatogram of PGNP obtained in example step 7;
FIG. 6 is a graph of the lgMw-RT (weight average molecular weight) calibration obtained in example step 8;
FIG. 7 is a chromatogram of PGNP obtained in example step 8;
FIG. 8 is a scanning ultraviolet spectrum of PGNP obtained in example step 9;
FIG. 9 is an infrared spectrum of PGNP obtained in example step 10;
FIG. 10 is a standard monosaccharide IC chromatogram obtained in example step 11;
FIG. 11 is a chromatogram of PGNPIC obtained in example step 11;
FIG. 12 is a graph of the body weight of mice obtained in step a of the example;
FIG. 13 is a view of HE section of liver tissue of mice obtained in step e of the example;
FIG. 14 is a chart showing the staining of the liver tissue with oil red O of the mice obtained in step f of the example.
Detailed Description
The technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Examples
1. The method for obtaining and verifying platycodon grandiflorum polysaccharide capable of improving constitution of mice after induction of high-fat diet comprises the following specific steps:
1. degreasing
Placing 50g of platycodon grandiflorum powder into a 500ml round-bottom flask, adding 250ml of absolute ethyl alcohol, refluxing in 80 ℃ water bath for 1h, repeating twice, suction filtering, taking filter residues, and drying in the shade to obtain defatted platycodon grandiflorum powder.
2. Water extraction
100g of defatted platycodon grandiflorum powder is placed in a 2L round-bottom flask, 1500ml of distilled water is added, the temperature is increased to 80 ℃, the water is extracted for 3 times, each time for 1.5 hours, 300-mesh nylon gauze is used for carrying out primary filtration on water extract, then suction filtration is carried out, and three filtrates are combined, heated and concentrated to obtain the target platycodon grandiflorum polysaccharide water extract.
3. Alcohol precipitation
Adding 95% ethanol into the target radix Platycodi polysaccharide water extract to reach alcohol concentration of 80%, standing at 4deg.C overnight, centrifuging (3500 rpm,10 min), and removing upper layer solution to obtain radix Platycodi polysaccharide-protein mixture.
4. Deproteinization and measurement of protein content
4.1 obtaining crude polysaccharide of platycodon grandiflorum
Distilled water is added into the platycodon polysaccharide-protein mixture to obtain a solution with the concentration of 1mg/4mL, and the volume ratio is 25:4:1, platycodon grandiflorum polysaccharide-protein mixture solution: chloroform: mixing n-butanol to obtain a mixed solution; and (3) carrying out electromagnetic stirring on the mixed solution for 30min, then carrying out centrifugation (4000 rpm,10 min), sucking the upper aqueous phase solution and keeping for standby, repeating the above operation for 12 times, mixing the 12 aqueous phase solutions, and carrying out freeze drying for 48 hours to obtain the platycodon grandiflorum crude polysaccharide.
4.2 measurement of protein content
The protein content in the platycodon grandiflorum crude polysaccharide is measured by adopting a Coomassie brilliant blue method, the result is shown in figure 1, and the measured protein content is as follows: 8.2%.
5. Determination of sugar content in platycodon grandiflorum crude polysaccharide
The content of the crude platycodon grandiflorum polysaccharide is measured by adopting a phenol-concentrated sulfuric acid method, the result is shown in figure 2, and the content of the sugar in the crude platycodon grandiflorum polysaccharide is as follows: 66.5%.
6. Cellulose column separation
35g of DEAE-52 cellulose (Soy Biotechnology Co., ltd.) was poured into 3500mL of degassed distilled water and soaked overnight, and the upper layer of suspended fine particles was poured off to prepare a column (2.6X 30 cm). 50mg of platycodon grandiflorum crude polysaccharide is dissolved in 5mL of distilled water, and the solution is filtered through a microporous membrane of 0.22 mu m, so as to prepare a sample. Slowly dripping the sample on the surface of the column packing, and eluting with distilled water twice the column volume after the sample fully fits the packing. The flow rate was 0.6mL/min, one tube was collected for 5min, and 76 tubes were collected in total. The phenol-concentrated sulfuric acid method is used for measuring the sugar content, and the elution curve is shown in figure 3. Concentrating 25-50 tubes of sugar solution, and lyophilizing to obtain radix Platycodi neutral polysaccharide.
The neutral polysaccharide sugar content of the platycodon grandiflorum is 92% by a phenol-concentrated sulfuric acid method.
7. Sephadex column separation
15g sephadex G100 (Soy Biotechnology Co., ltd.) was poured into 1500mL of degassed distilled water and soaked overnight, and the upper layer of suspended fine particles was poured off to prepare a column (1.6X 60 cm).
30mg of platycodon grandiflorum neutral polysaccharide purified by a DEAE-52 cellulose column is dissolved by 10mL of water, and the solution is filtered through a microporous membrane of 0.22 mu m to prepare a sample. Slowly dripping the sample on the surface of the column packing, fully attaching the sample to the packing, eluting with distilled water, collecting one tube at 0.4ml/min for 5min, and collecting 76 tubes in total. Detecting with phenol-sulfuric acid method, drawing elution curve, combining 25-60 tubes of eluates, concentrating, lyophilizing for 48 hr to obtain uniform radix Platycodi polysaccharide, and naming it as PGNP as shown in figure 4.
The resulting solution was dissolved in 2mg/mL PGNP solution and filtered through a 0.22 μm microporous filter. The uniformity was measured by high performance liquid chromatography, and the results are shown in FIG. 5. High performance liquid chromatography was equipped with a gel exclusion chromatography column (TSK G3000 PWXL). Reaction conditions: ultrapure water is used as a mobile phase, the flow rate is 0.5mL/min, the column temperature is 30 ℃, and the sample loading amount is 20 mu L. Polysaccharide was identified with a differential detector.
The PGNP obtained from fig. 5 is a homogeneous platycodon polysaccharide.
8. Molecular weight and purity determination of PGNP
Instrument:
TABLE 1
Materials: naCl, manufacturer ACROS, lot A0356762, cat# 139725000
Standard substance:
TABLE 2
The experimental steps are as follows:
8.1, reagent configuration: preparing 0.05M NaCl solution, ultrasonic treating for 10min, and filtering with 0.45 μm microporous membrane.
8.2, sample (PGNP) and standard solution preparation: the sample and standard were precisely weighed, the sample was prepared as a 5mg/ml solution, centrifuged at 12000rpm for 10min, the supernatant was filtered through a 0.22 μm microporous filter membrane, and the sample was transferred to a 1.8ml sample-in vial.
8.3, chromatographic method: the lgMw-RT (weight average molecular weight) calibration curve and PGNP chromatogram were obtained by high performance gel chromatography, and the results are shown in FIG. 6 and FIG. 7. Wherein lgMw-RT correction curve equation is: y= -0.2056x+12.69, r 2 =0.9948; combining fig. 6 and 7, PGNP was found to have a weight average molecular weight of 2313.
The chromatographic conditions are as follows: chromatographic column: BRT105-104-102 series gel column (8X 300 mm); mobile phase: 0.05M NaCl solution; flow rate: 0.6ml/min, column temperature: 40 ℃; sample injection amount: 20 μl; a detector: differential detector RI-10A.
9. Acquisition of PGNP UV spectrogram
Experimental materials: deionized water;
1mg/ml PGNP was prepared, and a UV spectrum scan at 200-800nm was shown in FIG. 8, and it was seen in FIG. 8 that no distinct characteristic absorbance peaks were present at 260nm and 280nm, indicating that PGNP contained no distinct proteins or nucleic acids.
10. Acquisition of PGNP Infrared Spectrometry
Grinding and extruding PGNP and potassium bromide powder (1:100) together into sheet, and frequency range of 4000-400cm -1 Spectral measurements were performed below, data were collected and transmittance (%) plotted as a function of wave number (cm-1), as shown in FIG. 9.
As can be seen from FIG. 9, the distance between 3000 and 3600cm -1 The wide frequency band of (C) indicates the stretching vibration of O-H (OH). 2930cm -1 About is C-H stretching strap, 1417cm -1 About is a C-O stretching belt, 1617cm -1 The absorption peak at this point is C=O stretching vibration at 1025cm -1 The strong absorption in the vicinity represents the glycosidic bond and C-O and C-C stretching vibrations in the pyranose structure.
11. Determination of the monosaccharide composition of PGNP by ion chromatography
11.1 reagent configuration:
15mM NaOH solution: 2.4g of 50% NaOH solution, 2L of water;
15mMNaOH&100mM NaOAC solution: 1.2g of 50% NaOH solution, 8.2g of NaOAC,1L of water.
11.2 preparation and calculation methods of standard solutions
7 monosaccharide standard substances (arabinose, galactose, glucose, xylose, mannose, fructose, glucosamine hydrochloride and N-acetyl-D glucosamine) are taken to prepare a standard mother solution.
And precisely preparing concentration standard substances from the monosaccharide standard solutions as mixed labels. According to the absolute quantitative method, the mass of the different monosaccharides is determined, and the molar ratio is calculated according to the molar mass of the monosaccharides.
11.3 sample preparation
20mg of the sample (PGNP) was precisely weighed into an ampoule, 2ml of 6M HCL was added and hydrolyzed at 105℃for 8 hours. Accurately sucking the acid hydrolysis solution, transferring to a pipe, blowing with nitrogen, drying, adding 5ml of water, mixing, sucking 100uL, adding 900uL of deionized water, and centrifuging at 12000rpm for 5min. The supernatant was analyzed by IC.
11.4 chromatography methods
Chromatographic column: dionexCarbopacTMPA20 (3×150); mobile phase: H2O; 15mMNaOHC:15mMNaOH&100mM NaOAC; flow rate: 0.3ml/min; sample injection amount: 5. Mu.L; column temperature: 30 ℃; a detector: an electrochemical detector.
11.5 the results of the experiments are shown in FIGS. 10 and 11, where the IC chromatogram of the standard monosaccharide is shown in FIG. 10 and the IC chromatogram of the PGNP is shown in FIG. 11.
From FIGS. 10 and 11, the monosaccharide composition and molar ratio of PGNP are shown in Table 3
TABLE 3 Table 3
It can be seen that PGNP consisted mainly of fructose, glucose (84.1%, 10.9%).
Second, effect of PGNP on high fat diet mice
a. Influence of platycodon polysaccharide PGNP on body weight of mice
The Kunming mice (purchased from Anhui Long Lin Co., ltd.) were divided into a blank group (ordinary feed model D12450, 10% of energy from fat), a model group (high fat feed D12492, 60% of energy from fat), and a PGNP group (high fat feed +300mg/kg PGNP), 10 animals each, each group was weighed and fed for 4 weeks, and the weight change was as shown in FIG. 12. From fig. 12, it can be seen that the body weight of the model group is significantly higher than that of the blank group, indicating that the high-fat diet has a significant induction effect on obesity of mice; the PGNP group mice show a gentle body weight increase trend, and the intervention of the PGNP is obtained to effectively inhibit the body weight increase induced by high fat; the PGNP has a certain effect of resisting obesity induced by high-fat diet.
Effects of PGNP on mouse fat accumulation
The epididymal fat of the mice obtained in step a was taken after the sacrifice and weighed, and the epididymal fat weights of the mice are shown in Table 4 below. The epididymal fat weight of the model group was significantly higher than (p < 0.001) the blank group, indicating that the mice would accumulate a lot of adipose tissue in vivo after induction with a high fat diet. The PGNP group is obviously lower than the model group (p < 0.001), and is consistent with the change trend of the body weight of the mice, so that the PGNP has good inhibition effect on fat accumulation of the mice induced by high-fat diet.
TABLE 4 Table 4
Note that: comparison of all groups to model group, P <0.001
Effects of PGNP on blood lipid levels in mice
The amounts of TC (total cholesterol) in the serum of each group of mice were determined using a biochemical kit, and the results are shown in table 5. The TC (p < 0.05) content in the serum of the mice in the model group is significantly higher than that of the mice in the blank group; the TC content in the serum of mice in the PGNP group (p < 0.05) was significantly reduced compared to the model group. The PGNP is shown to be capable of effectively reducing the rise of the total cholesterol in the serum of mice induced by a high-fat diet, and has potential preventive effects on obesity-related hyperglycemia, hyperlipidemia and the like.
TABLE 5
Note that: all groups compared to model group, P <0.05
Effects of PGNP on MDA and SOD in mouse serum
Oxidative stress indicators (MDA, SOD) in the serum of each group of mice were measured. The results are shown in Table 6, and compared with the blank group, the MDA content in the serum of the mice in the model group is obviously increased, and the PGNP can effectively inhibit the increase of MDA (p < 0.001) by intervention, so that the PGNP has a certain improvement effect on the obesity-induced oxidative stress. For SOD, there was no significant difference between groups of mice, indicating that PGNP had no significant effect in increasing antioxidant enzymes.
TABLE 6
Note that: comparison of all groups to model group, P <0.001
e. HE section of liver tissue
As a result of HE-slicing the liver tissue of each group of mice obtained in a, as shown in fig. 13, the liver tissue of the model group showed fatty vacuoles and cell boundaries were blurred as compared with the blank group. And the PGNP group has clear cell boundary, which shows that the PGNP has the effect of improving liver cell injury caused by liver lipid accumulation.
f. Liver tissue oil red O staining results
The liver tissues of the mice in the group obtained in the step a are subjected to liver tissue oil red O staining, the obtained results are shown in figure 14, cells in a blank group can grow in a near egg sphere shape, the edges are clear, the cytosol is rich, and red oil drops are not deposited at the edges of the cells. In the model group, a clear red lipid drop was seen in the cells surrounding the nucleus, indicating successful modeling. Morphological observation is further carried out through an optical microscope, compared with a model group, the PGNP group shows a gradually decreasing trend of red lipid droplets in cells, and the lipid droplets become smaller, the color becomes lighter and the trend of approaching to a blank group is pointed. The PGNP is suggested to be capable of effectively reducing the content of cell lipid droplets, and has a certain prevention and treatment effect on obesity.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (3)
1. A platycodon grandiflorum polysaccharide capable of improving constitution of mice after induction of high-fat diet, wherein the platycodon grandiflorum polysaccharide specifically comprises Ara, glcN, gal, glc, xyl, man and fri with a molar ratio of 1.1:0.4:0.4:10.9:2.2:0.9:84.1; the constitution of the mice is specifically fat accumulation of epididymis or cell lipid drop content in liver tissue;
the preparation method of the platycodon grandiflorum polysaccharide comprises the following steps:
step 1, degreasing
Pulverizing radix Platycodi decoction pieces to obtain radix Platycodi powder, placing 50g of radix Platycodi powder into 500ml round bottom flask, adding 250ml of absolute ethanol, refluxing in 80deg.C water bath for 1 hr, repeating twice, suction filtering the obtained product, collecting the residue, and drying in the shade to obtain defatted radix Platycodi powder;
step 2, water extraction
Placing 100g of defatted radix Platycodi powder into a 2L round bottom flask, adding 1500ml of distilled water, heating to 80 ℃, extracting with water for 3 times, 1.5h each time, initially filtering the water extract with 300 mesh nylon gauze each time, filtering again, preserving the filtrate, mixing the three filtrates, and concentrating under heating to obtain the target radix Platycodi polysaccharide water extract;
step 3, alcohol precipitation
Adding 95% ethanol into radix Platycodi polysaccharide water extract to reach alcohol concentration of 80%, standing at 4deg.C overnight, centrifuging, and removing upper layer solution to obtain radix Platycodi polysaccharide-protein mixture;
step 4, deproteinizing
Adding distilled water into the platycodon polysaccharide-protein mixture to prepare a solution with the concentration of 1mg/mL, mixing the platycodon polysaccharide-protein mixture solution, chloroform and n-butanol according to the volume ratio of 25:4:1 to obtain a mixed solution, then electromagnetically stirring the mixed solution for 30min, centrifuging, and freeze-drying the aqueous phase solution for 48h to obtain platycodon polysaccharide;
step 5, cellulose column separation
Soaking 35g of DEAE-52 cellulose in 3500mL of degassed distilled water overnight, pouring out fine particles suspended on the upper layer, and preparing for column packing; dissolving 50mg of crude platycodon grandiflorum polysaccharide after protein removal in 5mL of distilled water, and preparing to load after the crude platycodon grandiflorum polysaccharide is fully dissolved and passes through a microporous filter membrane of 0.22 mu m; slowly dripping a sample on the surface of the column packing, fully attaching the sample to the packing, eluting with distilled water with the volume twice that of the column, collecting a tube for 5min at the flow rate of 0.6 mL/min; mixing all the collected solutions, concentrating, and freeze-drying to obtain radix Platycodi neutral polysaccharide;
step 6, sephadex column separation
Pouring 15g of sephadex G100 into 1500mL of de-aerated deionized water, soaking overnight, pouring out the fine particles suspended on the upper layer, and preparing a column, wherein the specification of the column is 1.6 x 60cm; dissolving 30mg of neutral polysaccharide of radix Platycodi with 10mL of water, filtering with 0.22 μm microporous membrane, and loading; slowly dripping sample on the surface of column packing, fully attaching the column packing, eluting with distilled water, collecting one tube 5min, detecting each tube of eluent by phenol-sulfuric acid method, drawing corresponding elution curve, mixing the eluates, concentrating, freeze drying to obtain uniform radix Platycodi polysaccharide, and naming PGNP.
2. Use of the platycodon grandiflorum polysaccharide according to claim 1 in a diet or medicament that improves mice following induction of a high fat diet.
3. Use of the platycodon grandiflorum polysaccharide according to claim 1 in a diet or medicament for improving obesity-related hyperglycemia and hyperlipidemia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210198625.8A CN114478821B (en) | 2022-03-01 | 2022-03-01 | Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210198625.8A CN114478821B (en) | 2022-03-01 | 2022-03-01 | Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114478821A CN114478821A (en) | 2022-05-13 |
| CN114478821B true CN114478821B (en) | 2023-04-25 |
Family
ID=81483602
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210198625.8A Active CN114478821B (en) | 2022-03-01 | 2022-03-01 | Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114478821B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150047876A (en) * | 2013-10-25 | 2015-05-06 | 충북대학교 산학협력단 | Composition for maturation of dendritic cell comprising Polysaccharide isolated from platycodon gradiflorum |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101830999A (en) * | 2010-05-21 | 2010-09-15 | 王振宇 | Method for extracting platycodon root polysaccharide |
| CN102477103B (en) * | 2010-11-22 | 2014-09-10 | 中国科学院上海药物研究所 | Platycodon grandiflorum polysaccharide, and degradation product, preparation method and application thereof |
| CN103222998A (en) * | 2012-01-30 | 2013-07-31 | 苏州九龙医院有限公司 | Comprehensive extracting method for platycodin and polysaccharide |
| CN104292355B (en) * | 2014-11-11 | 2016-09-28 | 江苏华能药业有限公司 | A kind of extracting method of platycodon root polysaccharide |
| US11083767B2 (en) * | 2017-05-24 | 2021-08-10 | Kyungpook National University Industry-Academic Cooperation Foundation | Pharmaceutical composition comprising an extract of platycodon grandiflorum and method for preventing or treating of obesity using the same |
| CA2973962A1 (en) * | 2017-07-20 | 2019-01-20 | Kyungpook National University Industry-Academic C | Pharmaceutical composition comprising platycodon grandiflorum extract for preventing or treating obesity |
-
2022
- 2022-03-01 CN CN202210198625.8A patent/CN114478821B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150047876A (en) * | 2013-10-25 | 2015-05-06 | 충북대학교 산학협력단 | Composition for maturation of dendritic cell comprising Polysaccharide isolated from platycodon gradiflorum |
Non-Patent Citations (1)
| Title |
|---|
| Jing Song 等.Effects of neutral polysaccharide from Platycodon grandiflorum on high-fat diet-induced obesity via the regulation of gut microbiota and metabolites.《Frontiers in Endocrinology》.2023,全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114478821A (en) | 2022-05-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xie et al. | Antitumor and immunomodulatory activities of a water-soluble polysaccharide from Chaenomeles speciosa | |
| CN109400741B (en) | Separation and purification method of ganoderma lucidum spore polysaccharide | |
| CN106084087A (en) | A kind of preparation method of Fructus Trichosanthis polysaccharide | |
| CN110343185B (en) | Wine-processed polygonatum polysaccharide and application thereof in regulating spleen deficiency and immune function | |
| CN113717295A (en) | Eucommia acidic polysaccharide, extraction method and application of eucommia acidic polysaccharide in preparation of medicine for treating fatty liver | |
| CN113896807A (en) | Fresh rehmannia root polysaccharide and preparation method and application thereof | |
| CN114478821B (en) | Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof | |
| CN112062866B (en) | Hericium erinaceus selenium-rich polysaccharide and preparation method and application thereof | |
| CN109232756B (en) | Suaeda salsa polysaccharide extract and preparation method and application thereof | |
| CN108359021B (en) | Method for rapidly preparing flaxseed polysaccharide with antiviral and immunoregulatory activities | |
| CN115869343A (en) | Application of Shandong Ganoderma extracellular ethanol precipitate in preparing antitumor drugs | |
| CN113429442B (en) | Method for separating tectoridin and tectorigenin from water extraction residues of rhizoma et radix Sichuan blackberry lily | |
| CN108424469A (en) | Gorgon fruit kernel polysaccharide and separation and extraction method and application thereof | |
| CN113105567B (en) | Paecilomyces cicadae mannan and preparation and application thereof | |
| CN104945533B (en) | A kind of preparation method of active corn stigma holosaccharide | |
| CN111187309A (en) | Preparation process and application of four components in cistanche | |
| CN116515009B (en) | Dictyophora indusiata egg polysaccharide, extraction method and application thereof in pancreatic cancer resistance | |
| CN107041903B (en) | Integrated new method for processing and concocting radix Polygoni Multiflori Preparata in producing area | |
| CN109678981A (en) | A kind of preparation method of safflower polysaccharide, product and application | |
| CN115991795B (en) | Mugwort root polysaccharide and preparation method and application thereof | |
| CN116178580B (en) | Wine-processed Polygonatum sibiricum polysaccharide extract and preparation method thereof | |
| CN116693716A (en) | Radix tinosporae polysaccharide and preparation method and application thereof | |
| CN113069484B (en) | Preparation method of microcapsule preparation of anti-aging medicine | |
| CN118496390A (en) | Armillariella erythrorhizon purified polysaccharide and preparation method and application thereof | |
| CN118240101A (en) | Dendrobium candidum polysaccharide, medicine or health product containing dendrobium candidum polysaccharide and application of dendrobium candidum polysaccharide |
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 |