CN110812367A - Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes - Google Patents
Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes Download PDFInfo
- Publication number
- CN110812367A CN110812367A CN201911154295.7A CN201911154295A CN110812367A CN 110812367 A CN110812367 A CN 110812367A CN 201911154295 A CN201911154295 A CN 201911154295A CN 110812367 A CN110812367 A CN 110812367A
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- water
- polygonum cuspidatum
- drying
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 116
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 96
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 96
- 241001648835 Polygonum cuspidatum Species 0.000 title claims abstract description 85
- 235000018167 Reynoutria japonica Nutrition 0.000 title claims abstract description 85
- 206010012601 diabetes mellitus Diseases 0.000 title claims abstract description 50
- 239000003814 drug Substances 0.000 title claims abstract description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 87
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 60
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 53
- 239000000243 solution Substances 0.000 claims description 51
- 239000006187 pill Substances 0.000 claims description 35
- 239000012071 phase Substances 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 24
- 238000001035 drying Methods 0.000 claims description 19
- 238000003809 water extraction Methods 0.000 claims description 19
- 230000002829 reductive effect Effects 0.000 claims description 16
- 238000002156 mixing Methods 0.000 claims description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims description 13
- 238000002386 leaching Methods 0.000 claims description 12
- 239000000758 substrate Substances 0.000 claims description 12
- 238000005303 weighing Methods 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 238000002481 ethanol extraction Methods 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 9
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 9
- 239000008346 aqueous phase Substances 0.000 claims description 9
- 239000002826 coolant Substances 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 239000011347 resin Substances 0.000 claims description 9
- 229920005989 resin Polymers 0.000 claims description 9
- 238000002390 rotary evaporation Methods 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 7
- 239000003480 eluent Substances 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 239000008118 PEG 6000 Substances 0.000 claims description 6
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 claims description 6
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 claims description 6
- 238000007865 diluting Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 238000003260 vortexing Methods 0.000 claims description 6
- 238000001291 vacuum drying Methods 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 4
- 238000001694 spray drying Methods 0.000 claims description 4
- 239000006189 buccal tablet Substances 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims description 3
- AMTWCFIAVKBGOD-UHFFFAOYSA-N dioxosilane;methoxy-dimethyl-trimethylsilyloxysilane Chemical group O=[Si]=O.CO[Si](C)(C)O[Si](C)(C)C AMTWCFIAVKBGOD-UHFFFAOYSA-N 0.000 claims description 3
- 238000001704 evaporation Methods 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 239000011159 matrix material Substances 0.000 claims description 3
- 238000002844 melting Methods 0.000 claims description 3
- 230000008018 melting Effects 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 229940083037 simethicone Drugs 0.000 claims description 3
- 239000003826 tablet Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 2
- 239000000469 ethanolic extract Substances 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 239000006196 drop Substances 0.000 claims 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 abstract description 10
- 229940079593 drug Drugs 0.000 abstract description 6
- 238000011160 research Methods 0.000 abstract description 5
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 4
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 3
- 238000003255 drug test Methods 0.000 abstract description 2
- 239000002547 new drug Substances 0.000 abstract description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 46
- 239000008280 blood Substances 0.000 description 46
- 210000004369 blood Anatomy 0.000 description 46
- 239000008103 glucose Substances 0.000 description 46
- 241000700159 Rattus Species 0.000 description 25
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical group CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 25
- 241000699670 Mus sp. Species 0.000 description 20
- 230000000694 effects Effects 0.000 description 20
- 150000004804 polysaccharides Polymers 0.000 description 20
- 244000153955 Reynoutria sachalinensis Species 0.000 description 19
- 235000003202 Reynoutria sachalinensis Nutrition 0.000 description 19
- 235000020828 fasting Nutrition 0.000 description 14
- 238000000605 extraction Methods 0.000 description 12
- 238000003304 gavage Methods 0.000 description 9
- 230000007062 hydrolysis Effects 0.000 description 8
- 238000006460 hydrolysis reaction Methods 0.000 description 8
- 210000002700 urine Anatomy 0.000 description 8
- 230000037396 body weight Effects 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 239000012488 sample solution Substances 0.000 description 7
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 7
- KJCVRFUGPWSIIH-UHFFFAOYSA-N 1-naphthol Chemical compound C1=CC=C2C(O)=CC=CC2=C1 KJCVRFUGPWSIIH-UHFFFAOYSA-N 0.000 description 6
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 6
- HIMXGTXNXJYFGB-UHFFFAOYSA-N alloxan Chemical compound O=C1NC(=O)C(=O)C(=O)N1 HIMXGTXNXJYFGB-UHFFFAOYSA-N 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 6
- 230000001603 reducing effect Effects 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 5
- 238000005520 cutting process Methods 0.000 description 5
- 235000005911 diet Nutrition 0.000 description 5
- 230000037213 diet Effects 0.000 description 5
- 238000007410 oral glucose tolerance test Methods 0.000 description 5
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 5
- 208000004880 Polyuria Diseases 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 229960003105 metformin Drugs 0.000 description 4
- 206010036067 polydipsia Diseases 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 229960001052 streptozocin Drugs 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 206010020710 Hyperphagia Diseases 0.000 description 3
- 102000004877 Insulin Human genes 0.000 description 3
- 108090001061 Insulin Proteins 0.000 description 3
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003651 drinking water Substances 0.000 description 3
- 235000020188 drinking water Nutrition 0.000 description 3
- 230000003203 everyday effect Effects 0.000 description 3
- 230000037406 food intake Effects 0.000 description 3
- 229940125396 insulin Drugs 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 208000022530 polyphagia Diseases 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 208000016261 weight loss Diseases 0.000 description 3
- 230000004580 weight loss Effects 0.000 description 3
- 208000031648 Body Weight Changes Diseases 0.000 description 2
- 208000002705 Glucose Intolerance Diseases 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 206010022489 Insulin Resistance Diseases 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000012490 blank solution Substances 0.000 description 2
- 230000004579 body weight change Effects 0.000 description 2
- RSEOPLGIFDMYKN-UHFFFAOYSA-N ethanol;naphthalen-1-ol Chemical compound CCO.C1=CC=C2C(O)=CC=CC2=C1 RSEOPLGIFDMYKN-UHFFFAOYSA-N 0.000 description 2
- 235000012631 food intake Nutrition 0.000 description 2
- HYBBIBNJHNGZAN-UHFFFAOYSA-N furfural Chemical compound O=CC1=CC=CO1 HYBBIBNJHNGZAN-UHFFFAOYSA-N 0.000 description 2
- 230000003914 insulin secretion Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 229960004329 metformin hydrochloride Drugs 0.000 description 2
- OETHQSJEHLVLGH-UHFFFAOYSA-N metformin hydrochloride Chemical compound Cl.CN(C)C(=N)N=C(N)N OETHQSJEHLVLGH-UHFFFAOYSA-N 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 201000009104 prediabetes syndrome Diseases 0.000 description 2
- 238000011552 rat model Methods 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229940126673 western medicines Drugs 0.000 description 2
- SPFMQWBKVUQXJV-BTVCFUMJSA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanal;hydrate Chemical compound O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O SPFMQWBKVUQXJV-BTVCFUMJSA-N 0.000 description 1
- HSTZMXCBWJGKHG-UHFFFAOYSA-N (E)-piceid Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=CC(C=CC=2C=CC(O)=CC=2)=C1 HSTZMXCBWJGKHG-UHFFFAOYSA-N 0.000 description 1
- 235000020927 12-h fasting Nutrition 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 241000219050 Polygonaceae Species 0.000 description 1
- 241000205407 Polygonum Species 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000000337 buffer salt Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 230000009194 climbing Effects 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013118 diabetic mouse model Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 230000000214 effect on organisms Effects 0.000 description 1
- 230000007368 endocrine function Effects 0.000 description 1
- 238000004146 energy storage Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011049 filling Methods 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
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000037308 hair color Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 210000004153 islets of langerhan Anatomy 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002175 menstrual effect Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000465 moulding Methods 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
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003538 oral antidiabetic agent Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229960003764 polydatin Drugs 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 150000001629 stilbenes Chemical class 0.000 description 1
- 235000021286 stilbenes Nutrition 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- HSTZMXCBWJGKHG-CUYWLFDKSA-N trans-piceid Polymers O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=CC(\C=C\C=2C=CC(O)=CC=2)=C1 HSTZMXCBWJGKHG-CUYWLFDKSA-N 0.000 description 1
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 1
Images
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
- A61K31/715—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
- A61P3/08—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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
- A61K36/704—Polygonum, e.g. knotweed
Landscapes
- Health & Medical Sciences (AREA)
- Diabetes (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an application of polygonum cuspidatum polysaccharide in preparing a medicament for treating diabetes, and belongs to the technical field of traditional Chinese medicines. The inventor of the application discovers through drug test research that the polygonum cuspidatum polysaccharide has a good hypoglycemic effect in different type II diabetes models, so the polygonum cuspidatum polysaccharide can be used for preparing the drugs for treating diabetes, not only opens up new drugs for treating diabetes, but also opens up new application of the polygonum cuspidatum polysaccharide, and has good market application prospect.
Description
Technical Field
The invention relates to an application of polygonum cuspidatum polysaccharide in preparing a medicament for treating diabetes, belonging to the technical field of traditional Chinese medicines.
Background
Polysaccharides (Polysaccharides) are polymers formed by connecting monosaccharides, widely exist in animal cell membranes, plant and microbial cell walls, and are another important class of biomolecules in organisms besides nucleic acids and proteins. A large number of researches show that the polysaccharide has various functional characteristics, can be used as an energy storage substance, has a plurality of biological activities such as immunoregulation, tumor resistance, blood sugar reduction, blood fat reduction, radiation resistance, aging delay and the like, and has almost no toxic or side effect on organisms. At present, the chemical structure, pharmacological action, mechanism and structure-activity relationship of polysaccharide have become one of the most active fields in the life science research. Polysaccharides extracted from different organisms have different physiological effects.
Diabetes mellitus is a chronic metabolic disease caused by a defect in insulin secretion or action, and is mainly manifested by an increase in blood sugar level, and typical "more than three and one less" symptoms, i.e., polydipsia, polyphagia, polyuria and weight loss. Type II diabetes, also known as non-insulin dependent diabetes mellitus, refers to a type of diabetes mellitus in which insulin resistance is mainly accompanied by relative insufficiency of insulin or insufficient insulin secretion is mainly accompanied by insulin resistance, accounts for about 90% of the total number of diabetic patients, and develops slowly and insidiously. Most patients can stably control blood sugar after diet control and oral hypoglycemic drug treatment without depending on insulin. At present, most of the medicines for treating diabetes are western medicines, the toxic and side effects are relatively large, the toxic and side effects of the traditional Chinese medicines are less than those of the western medicines, the effects are relatively mild and lasting, and the traditional Chinese medicines have a relatively good comprehensive treatment effect and have certain advantages in the aspect of treating diabetes.
Polygonum Cuspidatum is a dried rhizome and root of Polygonum Cuspidatum (Polygonum Cuspidatum Sieb. et Zucc) which is a perennial herb of Polygonum of Polygonaceae, is recorded in Ben Cao gang mu and Dian nan Ben Cao and has the effects of promoting blood circulation, removing blood stasis, restoring menstrual flow, relieving cough, clearing heat, detoxifying and the like. The current research shows that the giant knotweed has the function of reducing blood sugar, and mainly plays roles in stilbenes, anthraquinones, flavonoids and the like. However, the prior art does not record whether the polygonum cuspidatum polysaccharide has the function of reducing blood sugar. If the polygonum cuspidatum polysaccharide has the function of reducing blood sugar, not only a new medicine for treating diabetes is developed, but also a new application of the polygonum cuspidatum medicinal material is developed, and the polygonum cuspidatum polysaccharide has great social significance and economic benefit.
Disclosure of Invention
The invention aims to provide application of polygonum cuspidatum polysaccharide. The inventor of the application discovers through drug test research that the polygonum cuspidatum polysaccharide has a good hypoglycemic effect in different type II diabetes models, so the polygonum cuspidatum polysaccharide can be used for preparing the drugs for treating diabetes, not only opens up new drugs for treating diabetes, but also opens up new application of the polygonum cuspidatum polysaccharide, and has good market application prospect.
The technical scheme for solving the problems is as follows: application of rhizoma Polygoni Cuspidati polysaccharide in preparing medicine for treating diabetes is provided.
The invention has the beneficial effects that:
the invention discovers that the giant knotweed polysaccharide can be used for preparing the medicine for treating the diabetes for the first time, not only opens up a new medicine for treating the diabetes, but also opens up a new application of the giant knotweed medicinal material, and has good market application prospect.
On the basis of the technical scheme, the invention can be further improved as follows.
Further, the medicine is one of granules, tablets, buccal tablets, capsules, pills and dropping pills.
The adoption of the further beneficial effects is as follows: the giant knotweed polysaccharide can be prepared into medicines with various dosage forms, is convenient for patients to take under different conditions, and is more flexible and convenient.
Further, the dripping pill is prepared from polygonum cuspidatum polysaccharide and a dripping pill substrate, wherein the mass ratio of the polygonum cuspidatum polysaccharide to the dripping pill substrate is 1:2, and the dripping pill substrate is prepared from PEG 6000 and PEG 4000 according to the mass ratio of 1: 2; the preparation method of the dripping pill comprises the following steps: (1) respectively weighing PEG 6000 and PEG 4000 according to the mass ratio of 1:2 to obtain a dripping pill substrate; (2) heating and melting dripping pill matrix in water bath at 80-85 deg.C, adding rhizoma Polygoni Cuspidati polysaccharide at a mass ratio of 1:2, and mixing; (3) controlling the temperature of coolant at 10 deg.C, dropping at 70-75 deg.C, and dropping distance at 9cm, collecting formed dripping pill, wiping off coolant on surface, wherein the coolant is simethicone, and drying.
The adoption of the further beneficial effects is as follows: there is no report about giant knotweed polysaccharide dropping pills in the prior art. After many experiments, the inventor finds out the preparation method of the polygonum cuspidatum polysaccharide dropping pill, and the obtained polygonum cuspidatum polysaccharide dropping pill is quickly disintegrated in vitro and is favorable for being absorbed in vivo after being orally taken, so that the curative effect is quickly exerted.
Further, the giant knotweed polysaccharide is prepared by adopting a water extraction and alcohol precipitation method, and comprises the following steps:
step 1.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml water, leaching at 90 deg.C for 3 times, each for 1 hr, mixing leaching solutions, filtering, rotary evaporating and concentrating to 40ml to obtain water extract concentrate;
step 1.2: removing protein in the water extraction concentrated solution obtained in the step 1.1 by a Sevag reagent method to obtain a protein-removed water extraction concentrated solution;
step 1.3: extracting the protein-removed water extraction concentrated solution obtained in the step 1.2 with chloroform of the same volume once, and taking a water layer;
step 1.4: adding 95 vol% ethanol into the water layer obtained in the step 1.3 until the concentration of the ethanol solution is 90 vol%, standing, precipitating, washing with acetone and diethyl ether for 2 times respectively, and drying to obtain crude polygonum cuspidatum polysaccharide;
step 1.5: diluting the crude polygonum cuspidatum polysaccharide obtained in the step 1.4 by 5-10 times with water, passing through D101 macroporous resin, eluting with distilled water until the eluted part is negative by Molish reaction detection, and obtaining water eluate;
step 1.6: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (1.5) to 20ml, and drying to obtain the refined polygonum cuspidatum polysaccharide.
The adoption of the further beneficial effects is as follows: the polygonum cuspidatum polysaccharide component can be extracted from polygonum cuspidatum by a water extraction and alcohol precipitation method, the extraction method is simple, the extraction rate is high, the operation is easy, the cost is low, and the method is suitable for large-scale popularization and application.
Furthermore, in step 1.2, the specific method of the Sevag reagent method is:
mixing the water extraction concentrated solution obtained in the step 1.1 with a Sevag reagent according to a volume ratio of 3:1, vortexing for 5min, centrifuging at 4000r/min for 1min to obtain a first water phase and a first chloroform phase, and separating the first water phase from the first chloroform phase; wherein the Sevag reagent is prepared by mixing chloroform and n-butyl alcohol according to the volume ratio of 4: 1;
adding 1/3 volumes of the Sevag reagent to the first aqueous phase, vortexing for 5min, centrifuging at 4000r/min for 1min to obtain a second aqueous phase and a second chloroform phase, and separating the second aqueous phase from the second chloroform phase;
combining the first chloroform phase and the second chloroform phase, adding water with the total volume of 1/4 volumes of the first chloroform phase and the second chloroform phase, washing for three times, separating, and combining the three washing liquids with the second water phase to obtain the protein-removed water extraction concentrated solution.
Further, in step 1.5, the elution rate is 0.5-1 mL/min.
Further, in step 1.4 and step 1.6, the drying manner is any one of vacuum drying, freeze drying, spray drying and reduced pressure drying.
Further, the polygonum cuspidatum polysaccharide is prepared by an ethanol extraction method, and comprises the following steps:
step 2.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml 66 vol% ethanol, leaching at 84 deg.C for 2 times, each for 1 hr, mixing the leaching solutions, and filtering to obtain ethanol extractive solution;
step 2.2: performing rotary evaporation and concentration on the ethanol extracting solution obtained in the step 2.1 until ethanol is evaporated out to obtain an ethanol extraction concentrated solution;
step 2.3: concentrating the ethanol extract concentrated solution obtained in the step 2.2 in water bath to obtain an extract;
step 2.4: diluting the extract obtained in the step 2.3 by 5-10 times with water, passing through D101 macroporous resin, eluting with distilled water until the eluate is negative by Molish reaction detection to obtain water eluate;
step 2.5: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (2.4) to 40ml, and drying to obtain the refined polygonum cuspidatum polysaccharide.
The adoption of the further beneficial effects is as follows: the polygonum cuspidatum polysaccharide component can be extracted from polygonum cuspidatum by an ethanol extraction method, the extraction method is simple, the extraction rate is high, the operation is easy, the cost is low, and the polygonum cuspidatum polysaccharide component is suitable for large-scale popularization and application.
Furthermore, in step 2.3, the relative density of the extract at 25 ℃ is 1.20.
Further, in step 2.5, the drying manner is any one of vacuum drying, freeze drying, spray drying and reduced pressure drying.
Drawings
FIG. 1 is a graph of the average water consumption per rat per group in example 6 of the present invention;
FIG. 2 is a graph showing the average urine output per rat in each group in example 6 of the present invention;
FIG. 3 shows the weight change of rats in each group in example 6 of the present invention;
FIG. 4 is a graph showing the effect of polydatin on blood glucose in rats in example 6 of the present invention;
FIG. 5 is a graph showing the body weight changes of the mice of each group after the gavage administration in example 6 of the present invention;
fig. 6 is a graph of the oral glucose tolerance test (x ± s, n ═ 6) in mice in example 6 of the present invention;
FIG. 7 shows the effect of polygonum cuspidatum polysaccharide on fasting plasma glucose in diabetic mice in example 6 of the present invention (x. + -.s, n. is 6).
Detailed Description
The principles and features of this invention are described below in conjunction with the following detailed drawings, which are given by way of illustration only and are not intended to limit the scope of the invention.
Example 1: preparation of giant knotweed polysaccharide
The giant knotweed polysaccharide is prepared by a water extraction and alcohol precipitation method, and comprises the following steps:
step 1.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml water, leaching at 90 deg.C for 3 times, each for 1 hr, mixing leaching solutions, filtering, rotary evaporating and concentrating to 40ml to obtain water extract concentrate.
Step 1.2: mixing the water extraction concentrated solution obtained in the step 1.1 with a Sevag reagent according to a volume ratio of 3:1, vortexing for 5min, centrifuging at 4000r/min for 1min to obtain a first water phase and a first chloroform phase, and separating the first water phase from the first chloroform phase; wherein the Sevag reagent is prepared by mixing chloroform and n-butyl alcohol according to the volume ratio of 4: 1.
Adding 1/3 volumes of the Sevag reagent to the first aqueous phase, vortexing for 5min, and centrifuging at 4000r/min for 1min to obtain a second aqueous phase and a second chloroform phase, and separating the second aqueous phase from the second chloroform phase.
Combining the first chloroform phase and the second chloroform phase, adding water with the total volume of 1/4 volumes of the first chloroform phase and the second chloroform phase, washing for three times, separating, and combining the three washing liquids with the second water phase to obtain the protein-removed water extraction concentrated solution.
Step 1.3: extracting the protein-removed water extraction concentrated solution obtained in the step 1.2 with chloroform of the same volume once, and taking a water layer.
Step 1.4: and (3) adding 95 vol% ethanol into the water layer obtained in the step (1.3) until the concentration of the ethanol solution is 90 vol%, standing, precipitating, washing with acetone and diethyl ether for 2 times respectively, and drying for 8-12 hours until the weight is not reduced or the water content is lower than 3% at the vacuum degree of-0.1 MPa and the temperature of 40-60 ℃ to obtain the crude polygonum cuspidatum polysaccharide.
Step 1.5: and (3) adding water to dilute the crude polygonum cuspidatum polysaccharide obtained in the step (1.4) by 5-10 times, passing through D101 macroporous resin, and eluting with distilled water until the eluted part is negative by Molish reaction detection, thus obtaining water eluate.
Wherein, the D101 macroporous resin needs to be pretreated, and the method comprises the following steps: (1) weighing about 260g of macroporous resin, placing the macroporous resin in a 1000ml beaker, adding a proper amount of 95 vol% ethanol, soaking for 24 hours, pouring the ethanol in the beaker as far as possible, adding new 95 vol% ethanol, and filling the column (the height of the column is 60cm, and the inner diameter is 4 cm). (2) Eluting with 95 vol% ethanol at flow rate of 2BV/h until the effluent and water are mixed at a ratio of 1:3 without turbidity. (3) Finally washing with water until no alcohol smell is clear. The column is ensured to be free of air bubbles on the whole, if the air bubbles exist, the air bubbles can be removed by tapping the outer wall with an ear washing ball, and the air bubbles are not removed to be re-filled. The column is placed in a stable place which is not easy to touch, and waiting for sample loading is carried out. Before loading, the liquid in the column was drained and left at a height of 2 cm. Closing the water-stop clamp, slowly loading the sample without flushing the liquid level, opening the water-stop clamp after the sample loading is finished, controlling the flow rate to be 2BV/h, eluting the liquid by using distilled water when the liquid is placed close to the bed surface of the column, and collecting water eluent until the eluted part is negative by Molish reaction detection.
The Molish reaction, it is the reaction of sample solution with 5% α -naphthol ethanol solution and concentrated sulfuric acid, produce purple ring reaction between two liquid levels, its principle is that the reaction sugar is dehydrated under the action of concentrated sulfuric acid or concentrated hydrochloric acid to form furfural and its derivative, then reacts with α -naphthol to form purple red complex, and form purple ring between the liquid level of sugar solution and concentrated sulfuric acid. the concrete operation is that 2 drops of 5% α -naphthol solution are added to 1ml of solution to be tested, then shaken, and about 1ml of concentrated sulfuric acid is slowly added along the tube wall, and the tube is slowly raised, and the test tube is cut up without shaking, and the purple ring appears at two liquid levels, so that it is proved that the polysaccharide is contained, α -naphthol ethanol solution preparation method, 5g of α -naphthol is dissolved with 95 vol% ethanol, and the volume is fixed to 100 ml.
Step 1.6: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (1.5) to 20ml, drying at the vacuum degree of-0.1 MPa and the temperature of 40-60 ℃ for 8-12h until the weight is not reduced or the water content is lower than 3%, and thus obtaining the refined polygonum cuspidatum polysaccharide.
Example 2: preparation of giant knotweed polysaccharide
The polygonum cuspidatum polysaccharide in the embodiment 1 is prepared by an ethanol extraction method, and comprises the following steps:
step 2.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml 66 vol% ethanol, leaching at 84 deg.C for 2 times, each for 1 hr, mixing the leaching solutions, and filtering to obtain ethanol extractive solution.
Step 2.2: and (3) performing rotary evaporation and concentration on the ethanol extracting solution obtained in the step (2.1) until ethanol is evaporated out to obtain an ethanol extraction concentrated solution.
Step 2.3: and (3) concentrating the ethanol extraction concentrated solution obtained in the step (2.2) in a water bath to obtain an extract with the relative density of 1.20 at 25 ℃.
Step 2.4: diluting the extract obtained in the step 2.3 by 5-10 times with water, passing through D101 macroporous resin, eluting with distilled water until the eluate is negative by Molish reaction, and obtaining water eluate.
Step 2.5: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (2.4) to 40ml, and performing vacuum drying to obtain the refined polygonum cuspidatum polysaccharide.
Example 3: drawing of glucose standard curve
(1) Preparing a solution: precisely weighing 26.6mg of glucose standard substance, placing the glucose standard substance in a 25ml measuring flask, adding water for dissolving and fixing the volume, accurately sucking 2.5ml, placing the glucose standard substance in the 25ml measuring flask, adding water for dissolving and fixing the volume to obtain a glucose standard solution (0.1064 mg/ml). 5.0167g of phenol is weighed, the phenol is put into a beaker, a small amount of water is added for dissolving, then the mixture is placed into a 100ml measuring flask, and water is added for diluting to the scale, thus obtaining 5 percent phenol solution.
(2) Drawing a standard curve: precisely sucking 0ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0ml of glucose standard solution into a test tube, respectively adding water to 1ml, sequentially adding 1ml of 5% phenol solution and 5ml of concentrated sulfuric acid, shaking, plugging, and shaking up by inverting at 180 ℃. Boiling in water bath for 30min, and cooling to room temperature. The above operations are repeated a total of three times. The absorbance value was measured at 490nm in a blank tube without standard solution, and the regression equation was calculated.
The glucose standard curve was obtained as: a is 8.0263C-0.0116, r is 0.996(a is absorbance, C is glucose concentration).
Example 4: determination of Polygonum cuspidatum polysaccharide
Measured by the phenol-sulfuric acid method. Preparing a solution with a certain concentration from the polysaccharide solution as a sample solution, measuring the absorbance of the sample solution by using an ultraviolet spectrophotometer, substituting the absorbance into a standard curve of glucose to obtain the concentration of the glucose, and calculating the mass of the polysaccharide. The method comprises the following specific operations: precisely sucking the prepared blank solution (distilled water) and 1ml of each sample solution, placing the blank solution and each sample solution into a 10ml test tube, adding 1ml of 5% phenol solution, shaking, adding 5ml of concentrated sulfuric acid, shaking, plugging, and shaking up by inverting at 180 ℃. Boiling in water bath for 30min, and cooling to room temperature. Absorbance was measured at 490 nm. The absorbance was substituted into the glucose standard curve of example 3 to obtain the glucose concentration.
Calculating the polysaccharide mass according to formula 1: polysaccharide mass (m) is 0.9nCV (equation 1). Wherein C is the concentration of glucose in the sample solution, n is the dilution multiple of the sample solution, V is the volume of the polysaccharide solution, and 0.9 is a conversion factor.
The hydrolysis conditions of the polysaccharide are considered, a proper amount of sample liquid is respectively measured and put into a test tube, 5% phenol solution is added, concentrated sulfuric acid is added, hydrolysis is carried out according to different conditions, ① is placed at room temperature for 40min, ② is placed at room temperature for 10min, 60 ℃ is carried out in water bath for 30min, cooling is carried out to room temperature, ③ is carried out at room temperature for 10min, 60 ℃ is carried out in water bath for 60min, cooling is carried out to room temperature, ④ is carried out at room temperature for 10min, boiling water bath is carried out for 30 min.
Specific data are shown in Table 1
TABLE 1 hydrolysis conditions of Polygonum cuspidatum polysaccharide results
As shown in Table 1, the hydrolysis conditions ② and ① are significantly different from the hydrolysis conditions ① in comparison with the above-mentioned conditions, wherein ② is the optimal hydrolysis condition, and the hydrolysis conditions are changed from hydrolysis of polysaccharides into monosaccharides due to the fact that the temperature is too high or the time is too long, so that ② is selected to hydrolyze polygonum cuspidatum polysaccharides.
Example 5: calculation of extraction yield of Polygonum cuspidatum polysaccharide
Calculating the extraction rate of the polygonum cuspidatum polysaccharide according to a formula 2: the extraction rate of polygonum cuspidatum polysaccharide is M/M multiplied by 100% (formula 2). Wherein M is the mass of the polysaccharide, and M is the mass of the original giant knotweed medicinal material.
The mass of the polysaccharide was calculated according to equation 1 of example 4. The extraction rate of polygonum cuspidatum polysaccharide was calculated according to equation 2 of example 5. Specific data are shown in table 2.
Example 1 the average extraction rate of polygonum cuspidatum polysaccharide obtained by water extraction and alcohol precipitation method is 2.79%. Example 2 the average extraction rate of polygonum cuspidatum polysaccharide obtained by ethanol extraction method was 3.78%. Therefore, the extraction rate of the polygonum cuspidatum polysaccharide obtained by the ethanol extraction method in the embodiment 1 is better than that of the polygonum cuspidatum polysaccharide obtained by the water extraction and alcohol precipitation method in the embodiment 2.
TABLE 2 results of extraction of Polygonum cuspidatum polysaccharide
Example 6: medicinal effect experiment of giant knotweed polysaccharide for reducing blood sugar
1-streptozotocin-induced rat diabetes model
1.1 method of modelling of diabetes
50 male SD rats are raised in cages with weight (180 +/-5) g and SPF animal room environment, and are freely fed with water and food at room temperature of 20-24 ℃ and humidity of 45-55%. After the common feed is fed for 5 weeks, Fasting Blood Glucose (FBG for short) is measured after Fasting for 24 hours without water prohibition, and the diabetes modeling is carried out. Randomized into type ii diabetes model group (DM) and normal control group (NC).
Injecting Streptozotocin (STZ) into abdominal cavity twice every other day according to 40mg/kg and 20mg/kg dose in type II diabetes model group (DM), continuously taking rat tail venous blood at the same time every day for 3 days after STZ is injected for 72 hours, and detecting fasting blood glucose by adopting a blood glucose analyzer. If the blood sugar is stably increased for 3 consecutive days and the fasting blood sugar is more than or equal to 11.1mmol/L, the molding is judged to be successful. Wherein, the STZ is prepared at present, dissolved in 0.1M citric acid buffer solution (pH value of 4.5), and stored in ice at low temperature in dark place before use.
The normal control group (NC) was injected with the same dose of citrate buffer.
1.2 administration by groups
SD rats injected with citric acid buffer salt are used as a normal control group, and diabetic rats successfully molded take blood sugar as a grouping factor, and are divided into four groups, namely a normal control group, a type II diabetes model control group, a metformin group and a polygonum cuspidatum polysaccharide group. The type II diabetes model control group and the normal control group are subjected to intragastric administration by using equivalent physiological saline, the metformin group is subjected to intragastric administration by using metformin suspension at a dose of 150mg/kg, the polygonum cuspidatum polysaccharide group is subjected to intragastric administration at a dose of 140mg/kg, the polygonum cuspidatum polysaccharide group is subjected to administration at a dose of 10:00 to 12:00 in the morning every day, and the administration is continuously performed by intragastric administration once a day according to the weight for 3 weeks.
1.3 results of the experiment
1.3.1 Observation of Water intake
The same rat diet was given to each rat on average daily, and diabetic rats typically had "more than three and one less", i.e., polydipsia, polyphagia, polyuria and weight loss. The water intake is observed in figure 1. As can be seen from FIG. 1, the water intake of the other groups was significantly increased as compared with the normal control group, which was consistent with the polydipsia phenomenon of diabetes. The metformin group and the polygonum cuspidatum polysaccharide group had slightly less water intake than the other groups. The water intake fluctuations of the groups of diabetes were not very different.
1.3.2 Observation of urine output
The urine output of the rats was received by the metabolism cages and the urine output was measured as shown in FIG. 2. As is apparent from fig. 2, in the normal control group, the average urine output per group was lower than that of the other groups under the condition of no significant difference in food intake, and the average urine output per group after administration was higher than that of the normal control group in the successfully molded groups, which was consistent with the diabetic polyuria phenomenon. Compared with the model control group, the urine volume discharged by the metformin group and the polygonum cuspidatum polysaccharide group is slightly reduced, and no significant difference exists.
1.3.3 rat body weight Change
During the experiment, changes in body weight of rats were monitored and recorded every 3 days. The body weight of the rats was recorded as in figure 3. As can be seen from FIG. 3, the body weight of the normal control group gradually increased and the body weight of each group with diabetes decreased under the condition of no obvious difference in food intake, which is in accordance with the weight decrease characteristics of diabetes. Compared with the model control group, the weight of the metformin group and the polygonum cuspidatum polysaccharide group is slightly increased, and no significant difference exists.
1.3.4 rat blood glucose assay
In the 0d, 3d, 6d, 9d, 12d, 15d, 18d and 21d of gavage polygonum cuspidatum polysaccharide, fasting for 12h in advance without water prohibition is performed, rat tail venous blood is taken, and fasting blood glucose is detected by a blood glucose analyzer, and the result is shown in fig. 4. The blood glucose in the model control group and the diabetes mellitus group were relatively high compared to the normal control group; the metformin group had a significant decrease in blood glucose (p <0.05) compared to the model control group. The blood sugar of rats in the polygonum cuspidatum polysaccharide group is obviously reduced (p is less than 0.05), and the effect of reducing the blood sugar is slightly weaker than that of the metformin group (p is more than 0.05).
1.3.5 determination of glucose tolerance in rats
Glucose Tolerance (Oral Glucose Tolerance Test, abbreviated OGTT) measurement: after the last administration, SD rats are fasted for 5 hours without water prohibition, and are subjected to intragastric administration of 2g/kg of glucose solution. The blood glucose values of SD rats were measured at 0h, 0.5h, 1h and 2h after intragastric administration of glucose, respectively. Calculate area under the curve (AUC) according to equation 3:
AUC [ mmol/(h.L) ] -0.5A + B + C +0.5D (equation 3). In the formula, A, B, C, D are blood glucose values of 0h, 0.5h, 1h and 2h, respectively. The results are shown in Table 3.
TABLE 3 sugar tolerance values
Note: compared with the normal control group at the same period,#P<0.05, in contemporaneous comparison with the model group,△P<0.05。
as can be seen from Table 3, the glucose tolerance level of the rats in the model control group was significantly increased (p <0.05) compared to the normal control group. Compared with the model control group, the glucose tolerance level of the rat of the polygonum cuspidatum polysaccharide group is obviously reduced (p is less than 0.05), and the effect is slightly weaker than that of the metformin group.
Therefore, through Streptozotocin (STZ) induction diabetes rat model, 140mg/kg of polygonum cuspidatum polysaccharide and 150mg/kg of metformin are administered according to the body weight, and after the administration for 21 days by gastric lavage, the water intake, the urine discharge and the body weight of the polygonum cuspidatum polysaccharide group, the metformin group and the model control group have no obvious change under the condition of no obvious ingestion difference. The blood glucose in each group of diabetes was relatively high compared to the normal control group; compared with the model control group, the blood sugar of rats in the polygonum cuspidatum polysaccharide group and the metformin group is reduced (p is less than 0.05), and the effect of the polygonum cuspidatum polysaccharide group is slightly weaker than that of the metformin group (p is more than 0.05).
The glucose tolerance of each group of diabetes was elevated compared to the normal control group; compared with the model control group, the glucose tolerance of the polygonum cuspidatum polysaccharide group and the metformin group is reduced (p is less than 0.05), and the effect of the polygonum cuspidatum polysaccharide group is slightly weaker than that of the metformin group (p is more than 0.05).
2 tetraoxypyrimidine induced mouse diabetes model
2.1 preparation of mouse diabetes model
40 Kunming mice are taken, are fed with water in a common diet for 72 hours after adaptive feeding, are fasted and freely drunk for 14 hours, are weighed, except sodium citrate buffer solution with the pH value of 4.5 such as normal control group intraperitoneal injection and the like, the rest mice are all intraperitoneally injected with alloxan solution (ALx160mg/kg body weight, sodium citrate buffer solution with the pH value of 4.5, and the injection volume is 0.2ml/10g), insulin is injected for 0.2U in an abdominal cavity after 15min, and 25% glucose water is freely drunk after alloxan is injected for 1 hour. After 72h of free diet drinking water, a diabetes model is determined by measuring the blood sugar value of a mouse with a glucometer, wherein the blood sugar value of the mouse is more than or equal to 11.1mmol/L after three days after the last administration (fasting for 4 hours before measurement, tail cutting and blood taking are carried out).
2.2 grouping and administration
The diabetes model mice are evenly and randomly grouped into a model control group, a giant knotweed polysaccharide group and a metformin group. Normal control group, without any treatment. And (3) feeding distilled water to the model control group, feeding a polygonum cuspidatum polysaccharide solution to the polygonum cuspidatum polysaccharide group, and feeding a metformin hydrochloride suspension solution to the metformin group, wherein the gavage is performed according to 200mg/kg, the gavage times are 1 time/day, and the gavage times are continuously 22 days. Freely eating and drinking water, observing the change of the hair color of the mice every day, and recording the change of the diet, the drinking water, the padding humidity and the weight. And randomly taking three mice from each group at the 5 th time of the gavage administration for an oral glucose tolerance test, and measuring the blood glucose by cutting the tail after the 0 th time, the 5 th time, the 10 th time, the 13 th time, the 16 th time, the 19 th time and the 22 th time of fasting without water prohibition.
2.2 general conditions in diabetic model mice
The pancreatic islets of the mice in the diabetes model are damaged, the secretion function of the mice is lost, and then the endocrine function is disturbed, so that the metabolism of nutrient substances of the body is influenced, and the weight is reduced. The general condition of diabetic mice is shown in Table 4, and the weight change of the mice is shown in FIG. 5. Compared with the normal control group, the model control group has the typical phenomenon of 'more than three and one less' of polydipsia, polyphagia, polyuria and weight loss. Compared with the model control group, the mice of the metformin group and the polygonum cuspidatum polysaccharide group have the characteristic symptoms of more than three and one less than three which are reduced after treatment, and have less weight change and are not reduced any more and tend to be stable compared with the weight of the mice of the model control group. Compared with a model control group, the metformin group and the giant knotweed polysaccharide group have sensitive response and basically no different activities such as fur luster, shrugging, hunch back, climbing and the like compared with a blank group.
Table 4 general conditions of diabetic mice (x ± s, n ═ 6)
2.3 oral glucose tolerance test in mice
After fasting and free drinking for 12h, weighing, measuring fasting blood glucose by cutting the tail, feeding glucose (2g/kg) by gavage, starting timing, and measuring blood glucose values for 0h, 0.5h, 1.0h and 2.0h by cutting the tail. The time of administration of glucose by gavage was plotted as the abscissa and the average value of blood glucose was plotted as the ordinate, to obtain fig. 6. As can be seen from fig. 6, the blood glucose of the gavage polygonum cuspidatum polysaccharide mice was significantly reduced (p <0.05) compared to the model control group at the time points of 0h, 0.5h, 1.0h and 2.0 h. Compared with the positive control drug, the giant knotweed polysaccharide group has the effect of inhibiting the blood sugar rise at 1.0h and 2.0h, which is slightly lower than that of the metformin group (p is more than 0.05). Therefore, the effect of the polygonum cuspidatum polysaccharide on improving the impaired glucose tolerance of the diabetic mice is obvious, but is lower than that of a positive control medicament.
2.4 change in fasting plasma glucose in model mice
After the administration of the drugs at 0h, 5h, 10h, 13h, 16h, 19h and 22d, respectively, and fasting for 8h without water prohibition, blood glucose was measured by tail cutting, and the administration time was plotted as abscissa and the mean value of blood glucose was plotted as ordinate, to obtain fig. 7. As can be seen in fig. 7, the fasting blood glucose of the model control group was significantly higher than that of the normal control group (p <0.05), indicating that the diabetes model was established; compared with a model control group, after the giant knotweed polysaccharide (200mg/kg) is continuously administered for 22 days, the blood sugar level of the giant knotweed polysaccharide group is remarkably reduced (p is less than 0.05); compared with the metformin group, the blood sugar reduction degree of the polygonum cuspidatum polysaccharide group is not significant (p > 0.05). Therefore, the polygonum cuspidatum polysaccharide can obviously improve the blood sugar level of diabetic mice, and has a slightly weaker effect than that of metformin.
The experimental result shows that the giant knotweed polysaccharide can improve the typical symptoms of diabetes such as more than three and one less than three of mice and the like to a certain extent; the effect of the polygonum cuspidatum polysaccharide on relieving the impaired glucose tolerance of the diabetic mice is obvious (p is less than 0.05), and is slightly lower than the hypoglycemic effect of the metformin (p is more than 0.05); the giant knotweed polysaccharide can reduce fasting blood glucose (p is less than 0.05) of diabetic mice, and the effect is slightly lower than that of metformin hydrochloride (p is more than 0.05).
In conclusion, the polygonum cuspidatum polysaccharide has obvious blood sugar reducing effect on diabetes caused by a mouse diabetes model caused by alloxan and a streptozotocin-induced diabetes rat model, is mild in effect, and has good application prospect and health care effect. Therefore, the giant knotweed polysaccharide can be used for preparing the medicine for treating diabetes. The medicine is one of granules, tablets, buccal tablets, capsules, pills and dropping pills.
Example 7: preparation of dripping pills
The dripping pill is prepared from polygonum cuspidatum polysaccharide and a dripping pill substrate, wherein the mass ratio of the polygonum cuspidatum polysaccharide to the dripping pill substrate is 1:2, and the dripping pill substrate is prepared from PEG 6000 and PEG 4000 according to the mass ratio of 1: 2; the preparation method of the dripping pill comprises the following steps: (1) respectively weighing PEG 6000 and PEG 4000 according to the mass ratio of 1:2 to obtain a dripping pill substrate; (2) heating and melting dripping pill matrix in water bath at 80-85 deg.C, adding rhizoma Polygoni Cuspidati polysaccharide at a mass ratio of 1:2, and mixing;
(3) controlling the temperature of coolant at 10 deg.C, dropping at 70-75 deg.C, and dropping distance at 9cm, collecting formed dripping pill, wiping off coolant on surface, wherein the coolant is simethicone, and drying. Detection shows that the polysaccharide content in each dripping pill is over 33.3%.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. Application of rhizoma Polygoni Cuspidati polysaccharide in preparing medicine for treating diabetes is provided.
2. The use of claim 1, wherein the medicament is one of granules, tablets, buccal tablets, capsules, pills and drop pills.
3. The use of claim 2, wherein the dripping pill is prepared from polygonum cuspidatum polysaccharide and a dripping pill substrate, wherein the mass ratio of the polygonum cuspidatum polysaccharide to the dripping pill substrate is 1:2, and the dripping pill substrate is prepared from PEG 6000 and PEG 4000 in the mass ratio of 1: 2; the preparation method of the dripping pill comprises the following steps: (1) respectively weighing PEG 6000 and PEG 4000 according to the mass ratio of 1:2 to obtain a dripping pill substrate; (2) heating and melting dripping pill matrix in water bath at 80-85 deg.C, adding rhizoma Polygoni Cuspidati polysaccharide at a mass ratio of 1:2, and mixing; (3) controlling the temperature of coolant at 10 deg.C, dropping at 70-75 deg.C, and dropping distance at 9cm, collecting formed dripping pill, wiping off coolant on surface, wherein the coolant is simethicone, and drying.
4. The use of claim 1, wherein the polygonum cuspidatum polysaccharide is prepared by a water extraction and alcohol precipitation method, comprising the following steps:
step 1.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml water, leaching at 90 deg.C for 3 times, each for 1 hr, mixing leaching solutions, filtering, rotary evaporating and concentrating to 40ml to obtain water extract concentrate;
step 1.2: removing protein in the water extraction concentrated solution obtained in the step 1.1 by a Sevag reagent method to obtain a protein-removed water extraction concentrated solution;
step 1.3: extracting the protein-removed water extraction concentrated solution obtained in the step 1.2 with chloroform of the same volume once, and taking a water layer;
step 1.4: adding 95 vol% ethanol into the water layer obtained in the step 1.3 until the concentration of the ethanol solution is 90 vol%, standing, precipitating, washing with acetone and diethyl ether for 2 times respectively, and drying to obtain crude polygonum cuspidatum polysaccharide;
step 1.5: diluting the crude polygonum cuspidatum polysaccharide obtained in the step 1.4 by 5-10 times with water, passing through D101 macroporous resin, eluting with distilled water until the eluted part is negative by Molish reaction detection, and obtaining water eluate;
step 1.6: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (1.5) to 20ml, and drying to obtain the refined polygonum cuspidatum polysaccharide.
5. The use according to claim 4, wherein in step 1.2, the specific method of the Sevag reagent method is:
mixing the water extraction concentrated solution obtained in the step 1.1 with a Sevag reagent according to a volume ratio of 3:1, vortexing for 5min, centrifuging at 4000r/min for 1min to obtain a first water phase and a first chloroform phase, and separating the first water phase from the first chloroform phase; wherein the Sevag reagent is prepared by mixing chloroform and n-butyl alcohol according to the volume ratio of 4: 1;
adding 1/3 volumes of the Sevag reagent to the first aqueous phase, vortexing for 5min, centrifuging at 4000r/min for 1min to obtain a second aqueous phase and a second chloroform phase, and separating the second aqueous phase from the second chloroform phase;
combining the first chloroform phase and the second chloroform phase, adding water with the total volume of 1/4 volumes of the first chloroform phase and the second chloroform phase, washing for three times, separating, and combining the three washing liquids with the second water phase to obtain the protein-removed water extraction concentrated solution.
6. The use according to claim 4, wherein in step 1.5, the elution rate is 0.5-1 mL/min.
7. The use according to claim 4, wherein in step 1.4 and step 1.6, the drying is performed by any one of vacuum drying, freeze drying, spray drying and drying under reduced pressure.
8. The use of claim 1, wherein the polygonum cuspidatum polysaccharide is prepared by an ethanol extraction method, comprising the following steps:
step 2.1: pulverizing rhizoma Polygoni Cuspidati, weighing 20g, adding 600ml 66 vol% ethanol, leaching at 84 deg.C for 2 times, each for 1 hr, mixing the leaching solutions, and filtering to obtain ethanol extractive solution;
step 2.2: performing rotary evaporation and concentration on the ethanol extracting solution obtained in the step 2.1 until ethanol is evaporated out to obtain an ethanol extraction concentrated solution;
step 2.3: concentrating the ethanol extract concentrated solution obtained in the step 2.2 in water bath to obtain an extract;
step 2.4: diluting the extract obtained in the step 2.3 by 5-10 times with water, passing through D101 macroporous resin, eluting with distilled water until the eluate is negative by Molish reaction detection to obtain water eluate;
step 2.5: and (3) performing rotary evaporation and concentration on the water eluent obtained in the step (2.4) to 40ml, and drying to obtain the refined polygonum cuspidatum polysaccharide.
9. The use of claim 8, wherein the polygonum cuspidatum polysaccharide is prepared by an ethanol extraction method, and in step 2.3, the relative density of the extract at 25 ℃ is 1.20.
10. The use according to claim 8, wherein in step 2.5, the drying is performed by any one of vacuum drying, freeze drying, spray drying and drying under reduced pressure.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911154295.7A CN110812367A (en) | 2019-11-22 | 2019-11-22 | Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911154295.7A CN110812367A (en) | 2019-11-22 | 2019-11-22 | Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110812367A true CN110812367A (en) | 2020-02-21 |
Family
ID=69558230
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911154295.7A Pending CN110812367A (en) | 2019-11-22 | 2019-11-22 | Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110812367A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101204428A (en) * | 2007-12-17 | 2008-06-25 | 苗明三 | Substance extracted from giant knotweed for diabetes mellitus and complication thereof |
| RU2409381C1 (en) * | 2007-07-31 | 2011-01-20 | Сиюань Хоспитал, Чайна Акэдеми Оф Чайниз Медикал Сайнсис | Pharmaceutical composition for blood sugar and fat control, its manufacturing and administration thereof |
| CN105777496A (en) * | 2016-03-08 | 2016-07-20 | 陈爱华 | Method for extracting resveratrol from rhizoma polygoni cuspidate and rhizoma polygoni cuspidate healthcare product |
-
2019
- 2019-11-22 CN CN201911154295.7A patent/CN110812367A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2409381C1 (en) * | 2007-07-31 | 2011-01-20 | Сиюань Хоспитал, Чайна Акэдеми Оф Чайниз Медикал Сайнсис | Pharmaceutical composition for blood sugar and fat control, its manufacturing and administration thereof |
| CN101204428A (en) * | 2007-12-17 | 2008-06-25 | 苗明三 | Substance extracted from giant knotweed for diabetes mellitus and complication thereof |
| CN105777496A (en) * | 2016-03-08 | 2016-07-20 | 陈爱华 | Method for extracting resveratrol from rhizoma polygoni cuspidate and rhizoma polygoni cuspidate healthcare product |
Non-Patent Citations (2)
| Title |
|---|
| 刘希全: ""虎杖多糖的提取、分离纯化及结构研究"", 《中国优秀硕士学位论文全文数据库（医药卫生科技辑）》 * |
| 马平恩: "虎杖多糖的提取工艺研究", 《医药导报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN115364170B (en) | Sugar metabolism regulator and preparation method and application thereof | |
| CN103285070A (en) | Preparation method and application of chickpea sprout extract dispersible tablet | |
| CN103417749A (en) | Preparing method of herba violae extract and application of herba violae extract in hypoglycemic agents | |
| CN104922176A (en) | Application of flos chrysanthemi indici extract | |
| CN105125566B (en) | The application of mannoglucan aldehydic acid oligosaccharides and derivative in treatment and/or prevention nephrosis medicine or health products is prepared | |
| CN1957999B (en) | Composition of Chinese traditional medicine, preparation method, and checking method | |
| CN101574377B (en) | Effective part of sweetberry jointfir, preparation method and application thereof in preparing medicament for treating gout disease | |
| CN101744844B (en) | Fulvic acid or sodium fulvate substance with hypoglycemic function | |
| CN101618096A (en) | Madder extract and preparation method and application thereof | |
| CN102716135B (en) | Lupenone prevents in preparation or treats the application in the product of diabetes | |
| CN113730464A (en) | New application of rhizoma coptidis pill, extract and pharmaceutical composition thereof and rhizoma coptidis pill product | |
| CN101167781A (en) | Orally-administered hypoglycemic sweet potato leaf single prescription traditional Chinese medicine and preparation method thereof | |
| CN110812367A (en) | Application of polygonum cuspidatum polysaccharide in preparing medicine for treating diabetes | |
| CN103432420B (en) | A kind of Chinese medicine composition for the treatment of diabetes and preparation method thereof and detection method | |
| CN102824423B (en) | Medicinal composition comprising albiflorin and arctiin and application | |
| CN101301374A (en) | Preparation of loquat leaf total flavones and functions thereof for reducing blood sugar and blood fat | |
| CN100584345C (en) | Distillage of Ardisia chinensis Benth of possessing function of antivirus, extraction method and application | |
| CN103432124B (en) | The application of oxymatrine in preparation control medicine for treating diabetic nephropathy | |
| CN105168300A (en) | Pharmaceutical composition for treating diabetes and preparation method thereof | |
| CN101829271A (en) | Chinese medicinal compound with function of treating diabetes and preparation method and application thereof | |
| CN101249129B (en) | Chinese medicine extract combination and medicine use thereof | |
| CN105560607A (en) | Preparation method and application of preparation having antithrombotic and anti-myocardial ischemic effects | |
| CN102698096B (en) | Composition of traditional Chinese medicine extracts and preparation method and application thereof | |
| CN110314160A (en) | Berbamine prevents and treats the application in medicine for treating diabetic nephropathy in preparation | |
| CN105434886A (en) | Dendrobium officinale root extractive with blood-sugar reducing effect and uses thereof |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200221 |