CN102641317B - Golden wave extract and application thereof in preparation of antidiabetic agent - Google Patents
Golden wave extract and application thereof in preparation of antidiabetic agent Download PDFInfo
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- CN102641317B CN102641317B CN201210141831.1A CN201210141831A CN102641317B CN 102641317 B CN102641317 B CN 102641317B CN 201210141831 A CN201210141831 A CN 201210141831A CN 102641317 B CN102641317 B CN 102641317B
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Abstract
The invention discloses an extract containing marein, golden wave chalcone and coreopsin obtained by extracting golden wave in backflow mode which is heated through carbinol, golden wave or an extract containing marein and golden wave chalcone and free of coreopsin obtained by extracting and heating by water. The golden wave extract is externally provided with DPPIV restraining activity, is capable of improving sugar tolerance of 2 type diabetes mellitus mice, is capable of reducing fasting blood-glucose, and is better in medicine effect compared with extracts without coreopsin.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to extract and there is diabetes active drug composition from Coreopsis basalis.
Background technology
Coreopsis basalis (Coreopsis tinctoria Nuff.) has another name called snow chrysanthemum, is the dry capitulum of Compositae (compositae) golden pheasant Chrysanthemum (Coreopsis), single-lobe, polyphyll or half polyphyll, and ligulate flower is yellow or golden yellow, florescence 6～JIUYUE.Flos Caryophylli Coreopsis basalis belongs to north temperate zone plant, originate in North America, Florida Zhou Ji Mexico state, China 1936 introduces a fine variety the son ground cultivation such as to Mount Lushan, the Southern Mountain as ornamental plant, various places are except confession is viewed and admired at present, also use all herbal medicine, and the food coloring of extraction from spend, there is the effect of heat-clearing and toxic substances removing and blood pressure lowering.Xinjiang Coreopsis basalis is distributed widely in the Kunlun Mountains of 3000 meters of left and right of Hotan Prefecture's height above sea level, has more rich wild resource, among the people, often its dry flower is used for making tea.
Summary of the invention
The object of the invention is to extract and there is diabetes active drug composition from Coreopsis basalis.
It is as follows that the present invention realizes the technical scheme that above-mentioned purpose adopts:
Coreopsis extract, its extracting method is: Coreopsis basalis head inflorescence is pulverized, heating and refluxing extraction in the methanol that is 60～80% in mass concentration, temperature is 50～70 ℃, the methanol that concentrating under reduced pressure reclaims in extracting solution obtains fluid extract, fluid extract is extracted with ethyl acetate, and extract obtains coreopsis extract through concentrated, dry, contains marein, Coreopsis basalis chalcone and coreopsin in extract.
The mass ratio of the methanol that further, Coreopsis basalis head inflorescence powder and mass concentration are 60～80% during reflux, extract, is 1:(25～30).
Another kind of coreopsis extract, its extracting method is: Coreopsis basalis head inflorescence is pulverized, heating and refluxing extraction in water, temperature is 60-80 ℃, and concentrating under reduced pressure obtains fluid extract, polyamide column on fluid extract, the ethanol that is 30～95% by mass concentration carries out gradient elution, eluent obtains coreopsis extract through concentrated, dry, in extract, containing marein and Coreopsis basalis chalcone, does not contain coreopsin.
Further, the mass ratio of Coreopsis basalis head inflorescence powder and water is 1:(25～30 during reflux, extract).
Further, the application of above-mentioned coreopsis extract in preparing antidiabetic medicine.
Described marein has following structural formula:
；
Described Coreopsis basalis chalcone has following structural formula:
；
Described coreopsin has following structural formula:
。
Beneficial effect: in plant, the extraction of active drug composition adopts alcohol extraction or two kinds of methods of water extraction conventionally, the present invention finds all can from Coreopsis basalis, extract containing marein and Coreopsis basalis chalcone with methanol or water, compare with water extract, while using methanol extraction, extract is also containing coreopsin, prepared ethanol extract or water extract have carried out effect experiment to type 2 diabetes mellitus mice, result shows that coreopsis extract can improve the carbohydrate tolerance of type 2 diabetes mellitus mice, and there is the effect that reduces fasting glucose, in extract, contain coreopsin than the drug effect containing coreopsin is not better, the external effect with inhibition DPPIV of coreopsis extract, it falls hypoglycemic mechanism and may act on therewith relevant.
The specific embodiment
Below in conjunction with embodiment, the invention will be further described.
The chemical composition of extract adopts LC-MS instrument (LC/MS) to carry out qualitative analysis.
Embodiment 1
Xinjiang Coreopsis basalis head inflorescence is through pulverizing 20-50 mesh sieve, the methanol that is 60-80% by mass concentration, 50-70 ℃ reflux, extract, 2 times, and each 2h, feed liquid mass ratio is 1:25, merges the extracting solution of 2 times, concentrating under reduced pressure recovery solvent obtains fluid extract.Equal-volume ethyl acetate extraction 1-3 time for methanol extract fluid extract, combining extraction liquid, after Rotary Evaporators is concentrated, obtains ethyl acetate extraction extractum; Thermostatic drying chamber is dry, obtains crocus powder.
Embodiment 2
Xinjiang Coreopsis basalis head inflorescence, through pulverizing 20-50 mesh sieve, adds water, 60-80 ℃ of reflux, extract, 2 times, and each 2h, feed liquid mass ratio is 1:25, merges the extracting solution of 2 times, concentrating under reduced pressure obtains fluid extract.Polyamide column on water extract fluid extract, the ethanol that is 30%-95% by mass concentration carries out gradient elution, collects eluent, concentrated through Rotary Evaporators, and thermostatic drying chamber is dry, obtains brown ceramic powder.
The evaluation of embodiment 3 extract components
Precision takes each 10mg of extract that embodiment 1 and embodiment 2 make, and is placed in respectively 10ml volumetric flask, adds that methanol is ultrasonic dissolves it, is settled to scale, and filter membrane (0.22 μ m) filters twice, standby.Precision takes marein reference substance, coreopsin reference substance and Coreopsis basalis chalcone reference substance 4.69mg, 4.97mg and 4.82mg respectively, with chromatograph dissolve with methanol and be settled in 10mL volumetric flask, obtain the standard solution that concentration is respectively 0.469mg/mL, 0.497mg/mL and 0.482 mg/mL.LC/MS chromatographic condition: chromatographic column: HyPersil BDS C
18(4.6mm * 250nm, 5 μ m); Chromatographic peak acquisition range: 190nm～400nm; Mobile phase: methanol (A)-0.5%(quality) formic acid solution (B), gradient elution program sees the following form; Flow velocity: 0.8mlmin
-1; Column temperature: 25 ℃; Sample size: 10 μ l; Detect wavelength: 290nm; MS condition: the full ion scan of 100-800 karyoplasmic ratio.
。
LC/MS result shows, contains marein, Coreopsis basalis chalcone and three kinds of working substances of coreopsin during embodiment 1 is extract obtained, contains marein and Coreopsis basalis chalcone, without coreopsin during embodiment 2 is extract obtained.
Embodiment 4 pharmacodynamic experiments
One, the foundation of type 2 diabetes mellitus mouse model
140 of clean level male mouse of kunming, body weight 18-22g, wherein as Normal group normal feedstuff, feed for 10, as model group, use high glucose and high fat forage feed one week for another 130, after fasting 8 hours, lumbar injection streptozotocin (STZ solution) 180mg/kg(Normal group gives the aseptic citric acid-sodium citrate buffer of same volume).After 72 hours, fasting 12 hours, carries out 2h carbohydrate tolerance test (OGTT), blood glucose >=11.1mmol/L person modeling success.
Two, become mould mice group and administration
Become the grouping of mould mice
Get into 88 of mould mices, according to body weight, be divided at random 8 groups: heavy dose of group of extract obtained minute of model group, metformin positive control drug group, embodiment 1, middle dosage group, small dose group, heavy dose of group of extract obtained minute of embodiment 2, middle dosage group, small dose group, add totally 9 groups of aforementioned Normal groups, every group guarantees more than 10.
Become the administration of mould mice
Each group is in every morning gavage, and 1 times/day, normal group and model group are filled with normal saline, successive administration 30 days.Mice weekly tail point is got a blood survey fasting glucose (FBG) and oral glucose tolerance experiment (OGTT).
Three, experimental result
Before administration, model group and each administration group FBG and OGTT there was no significant difference.From administration, 2 weeks, start, the FBG of metformin group and OGTT continue to reduce obviously, and the heavy dose of group FBG of embodiment 1 and OGTT also reduce; After administration 4 weeks, embodiment 1 heavy dose of group FBG and OGTT and model group relatively have notable difference (
p<0.01), and in embodiment 1 dosage group FBG and OGTT and model group more also have significant difference (
p<0.05), embodiment 2 heavy dose of with middle dosage group FBG and OGTT and model group have significant difference (
p<0.05), relatively indifference is different in nature for embodiment 1 small dose group, embodiment 2 small dose group FBG and OGTT and model group.Concrete outcome is as follows:
The external impact on DPP IV (DPP IV) of embodiment 5 coreopsis extracts
By the dried powder of two kinds of coreopsis extracts of embodiment 1 and embodiment 2 gained, be mixed with the solution of variable concentrations gradient.Experiment is divided into 4 groups, establish 3 multiple holes for every group, be respectively blank group (buffer+substrate), negative control group (enzyme+buffer+substrate), experiment blank group (extract+buffer+substrate) (extract is colored substance, there will be self color interference while detecting at 405nm place, and the blank that therefore every kind of each concentration of extract is set when detecting is avoided false positive phenomenon as far as possible), experimental group (extract+enzyme+buffer+substrate), the suppression ratio of observing variable concentrations changes.First by each extract, DPPIV, substrate and buffer water-bath 30min at 37 ℃ of temperature, then according to the order of extract, enzyme, buffer, substrate, add successively in 96 orifice plates, addition is as table 1; Hatch 60min for 37 ℃, under 405nm wavelength, record OD value.
Experimental result is as follows:
More than experiment shows that coreopsis extract can improve the carbohydrate tolerance of type 2 diabetes mellitus mice and have the effect that reduces fasting glucose, wherein, the effect that embodiment 1 makes extract is better than the effect that embodiment 2 makes extract, illustrate in extract and contain coreopsin than the drug effect containing coreopsin is not better, two kinds of external effects with inhibition DPPIV of coreopsis extract, it falls hypoglycemic mechanism and may act on therewith relevant.
Claims (3)
1. a diabetes coreopsis extract, it is characterized in that, this extract is to obtain by following extracting method: Coreopsis basalis head inflorescence is pulverized, heating and refluxing extraction in the methanol that is 60～80% in mass concentration, temperature is 50～70 ℃, and the methanol that concentrating under reduced pressure reclaims in extracting solution obtains fluid extract, and fluid extract is extracted with ethyl acetate, extract obtains coreopsis extract through concentrated, dry, contains marein, Coreopsis basalis chalcone and coreopsin in extract.
2. coreopsis extract according to claim 1, is characterized in that: the mass ratio of the methanol that during reflux, extract,, Coreopsis basalis head inflorescence powder and mass concentration are 60～80% is 1:(25～30).
3. the application of the coreopsis extract described in claim 1 to 2 any one in preparing antidiabetic medicine.
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103222996A (en) * | 2013-05-15 | 2013-07-31 | 顾海 | Coreopsis tinctoria refined extract and medical application thereof |
| CN103989691A (en) * | 2014-02-27 | 2014-08-20 | 中国医学科学院药用植物研究所 | Application of marein in preventing and treating cognition impairments induced by pyruvic aldehyde pyruvaldehyde |
| CN104173400B (en) * | 2014-07-02 | 2018-01-02 | 苏州大学 | A kind of coreopsis tinctoria extract and its application |
| CN104098624B (en) * | 2014-07-29 | 2016-04-13 | 新疆农业大学 | The preparations and applicatio of a kind of mating type the Kunlun snow chrysanthemum phenyl styryl ketone glycoside compounds |
| CN107648294A (en) * | 2017-11-13 | 2018-02-02 | 新疆维吾尔自治区药物研究所 | The purposes in treating vascular dementia medicine is being prepared containing the double-colored golden wave active component of marein |
| CN108929209B (en) * | 2018-08-01 | 2021-11-09 | 新疆医科大学 | Method for extracting ocainin from coreopsis tinctoria |
| CN109010406A (en) * | 2018-08-20 | 2018-12-18 | 上海拉德钫斯生物科技有限公司 | A kind of preparation method for the coreopsis tinctoria extract improving diabetic nephropathy |
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| CN102048786A (en) * | 2010-12-09 | 2011-05-11 | 新疆维吾尔自治区药物研究所 | Preparation and application of coreopsis extract |
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