CN101632684A - Application of gynostemma pentaphylla polysaccharide in preparing medicine or health-care product preventing and treating senile dementia - Google Patents
Application of gynostemma pentaphylla polysaccharide in preparing medicine or health-care product preventing and treating senile dementia Download PDFInfo
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- CN101632684A CN101632684A CN200910017269A CN200910017269A CN101632684A CN 101632684 A CN101632684 A CN 101632684A CN 200910017269 A CN200910017269 A CN 200910017269A CN 200910017269 A CN200910017269 A CN 200910017269A CN 101632684 A CN101632684 A CN 101632684A
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Abstract
The invention discloses a novel application of gynostemma pentaphylla polysaccharide, namely an application of gynostemma pentaphylla polysaccharide in preparing a medicine or a health-care product preventing and treating senile dementia. When being applied, the gynostemma pentaphylla polysaccharide can be used as an active component to prepare a composition, and the form of the composition can be any one of injection, an oral preparation and an externally-applied preparation, preferably the oral preparation. Experiments show that the gynostemma pentaphylla polysaccharide has an obvious protective function to nerve cell beta-amyloid injuries and has obvious preventing and treating functions to a rat model with senile dementia. The experiments further show that the gynostemma pentaphylla polysaccharide has a favorable improving function to the pathologic change and the pathologic and physiological changes of the senile dementia and has favorable protection and restoration functions to nerve cell injuries, and foreshow that the gynostemma pentaphylla polysaccharide has a very large development and application prospect in preparing the medicine or the health-care product preventing and treating the senile dementia.
Description
Technical field
The present invention relates to the application of Herb Gynostemmae Pentaphylli polysaccharides in preparation control medicine for senile dementia or health product.
Background technology
Senile dementia, claim Alzheimer (Alzheimer ' s disease again, AD), it is a kind of PDCD that occurs in geratic period and presenium, refer to a kind of persistence higher nerve functional activity obstacle, promptly there be not under the state of disturbance of consciousness the obstacle of aspects such as memory, thinking, analytical judgment, visual space identification, emotion.Its characteristic pathological change is the cerebral cortex atrophy, and with amyloid-beta (β-amyloid, β-AP) deposit, neurofibrillary tangles (neurofibrillarytangles, NFT), mass memory neuron number reduces, and senile plaque (senileplaque, formation SP).Still the medicine that does not have at present specific treatment or reverse progression of disease.Also be one of four big common neurodegenerative diseases (other three kinds are Huntington's disease, Lu Jialeishi disease, parkinson).
Aging trend allows alzheimer disease patient quantity increase development along with modern medicine, and increasing old man youthens and has been full of vigor.But please do not forget: in the near future, China will enter aging society.Very important problem of simultaneous---the sickness rate of senile dementia is increasing year by year.
Investigation is found: the mean age that northern China is suffered from senile dementia is 75,76 years old, and the age of suffering from vascular dementia is many about 68 years old.The ratio of suffering from the severe senile dementia among the over-65s crowd reaches more than 5%, and by 80 years old, this ratio just rose to 15-20%.
Senile dementia is the pertinacious disease that influences the senior health and fitness.The data that U.S.'s " general psychiatry document " provides shows that the whole world has 10% to suffer from this disease approximately among the old people more than 60 years old.According to World Health Organization (WHO) and international senile dementia association statistics, the whole world has 1,800 ten thousand old men of surpassing to suffer from senile dementia at present.Expect the year two thousand twenty, global patients of senile dementia will reach 3,400 ten thousand people.
Herb Gynostemmae Pentaphylli Gynostemma pentaphyllum (Thunb) Makino, English name Fiveleaf Gynostemma Herb, for the perennial climbing shrub of Cucurbitaceae (Gucurbitceae) Gynostemma (Gynastemma), have another name called Herba Gynostemmatis, Herb Gynostemmae Pentaphylli.There are 16 kinds and 3 mutation in the whole world, and there are 15 kinds and 3 mutation in China, China's gynostemma plant aboundresources, and most the kind do not furtherd investigate as yet and utilized.Herb Gynostemmae Pentaphylli is the treasure of China's Chinese medicine, have the title of " southern Radix Ginseng ", is known as " Gospel grass " in Japan, one of four big health care medicines that to be the world rise from the nineties.
For dietotherapeutic, think by the traditional Chinese medical science with herb for Herb Gynostemmae Pentaphylli, Herb Gynostemmae Pentaphylli bitter in the mouth, cold in nature, the function of have heat-clearing and toxic substances removing, QI invigorating, cough-relieving, eliminating the phlegm.China application Herb Gynostemmae Pentaphylli among the people lengthens one's life and keeps fit with a long historyly, and the producing region masses adopt its stem and leaf in summer and fry in shallow oil water and make refreshment drink.Modern pharmacological research and clinical practice prove that Herb Gynostemmae Pentaphylli is being regulated immunity of organisms, mutation; antitumor, antioxidation is regulated the trophic centre nerve; many aspects such as cardiovascular protection have biological activity preferably, and Herb Gynostemmae Pentaphylli extract is oral does not have a toxic and side effects.
Present research to Herb Gynostemmae Pentaphylli mainly concentrates on saponin constituent wherein, and less to the Herb Gynostemmae Pentaphylli STUDY ON POLYSACHAROSE, does not see the relevant report that Herb Gynostemmae Pentaphylli polysaccharides is used for the treatment of senile dementia.
Summary of the invention
At above-mentioned prior art, the invention provides a kind of new purposes of Herb Gynostemmae Pentaphylli polysaccharides, i.e. the application of Herb Gynostemmae Pentaphylli polysaccharides in preparation control medicine for senile dementia or health product.
During application, can be that active component is made compositions with the Herb Gynostemmae Pentaphylli polysaccharides, the form of compositions can be in injection, oral formulations, the external preparation any.The preferred oral preparation.
During application, can be according to the specific requirement of different preparations, Herb Gynostemmae Pentaphylli polysaccharides goods with a certain amount of pure product Herb Gynostemmae Pentaphylli polysaccharides or certain purity are prepared into various pharmaceutical compositions as active component, be aided with pharmaceutically acceptable various carrier, these pharmaceutically acceptable carriers can be the various carriers that are widely used in the prior art in the medicine.Described pharmaceutical composition can prepare by the known method of prior art such as mixing, granulation, tabletting.Described pharmaceutical composition can also comprise the optional helper component that various pharmacy are used, as flavouring agent, coloring agent, sweeting agent etc.
After testing, Herb Gynostemmae Pentaphylli polysaccharides is to be the heteropolysaccharide of key component with rhamnose, xylose, arabinose, galactose.The present inventor utilizes Herb Gynostemmae Pentaphylli polysaccharides to carry out cell and zoopery, and experimental result shows: damage has significant protective effect to Herb Gynostemmae Pentaphylli polysaccharides to the neurocyte amyloid-beta, and the senile dementia mouse model is had tangible preventive and therapeutic effect.Further show; Herb Gynostemmae Pentaphylli polysaccharides has good improvement effect to pathology, the pathophysiological change of senile dementia; neural cell injury is had excellent protection and repair, indicating that Herb Gynostemmae Pentaphylli polysaccharides has very big development prospect in preparation control medicine for senile dementia or health product.
The present invention has obtained Herb Gynostemmae Pentaphylli polysaccharides GPS-3 on the basis of the separation and purification of Herb Gynostemmae Pentaphylli polysaccharides having been carried out system, and has studied its preventive and therapeutic effect to senile dementia, for the application of Herb Gynostemmae Pentaphylli polysaccharides in the senile dementia disease provides experimental basis.
The specific embodiment
The present invention is further illustrated below in conjunction with embodiment:
Embodiment 1: extract Herb Gynostemmae Pentaphylli polysaccharides, step is as follows:
After the fragmentation of Herb Gynostemmae Pentaphylli medical material, take by weighing 200g, add the 1000ml alcohol reflux twice, each 1h filters and volatilizes ethanol, adds 2L water, 100 ℃ are extracted 3 times, and each 0.5h merges supernatant, concentrating under reduced pressure, the centrifugal precipitation of removing, supernatant adds 3 times of volume of ethanol, 4 ℃ are spent the night, centrifugal, and precipitation is respectively washed once with ethanol, dehydrated alcohol, acetone, 50 ℃ of dryings 5 hours, Herb Gynostemmae Pentaphylli crude polysaccharides 12.8g, be brown-black powder shape solid.
It is 5% aqueous solution that the dissolving of above-mentioned crude polysaccharides is made into mass concentration, adds anhydrous calcium chloride to 2%, stirring at room 2h, and the centrifugal then Calcium Pectate precipitation of removing, supernatant adds H
2O
2, make H
2O
2Final concentration to 5% (mass volume ratio), regulate pH to 8.5-9 with sodium hydroxide solution, stirring at room 5h, centrifugal, abandon precipitation, supernatant adds 3 times of volume of ethanol, and 4 ℃ are spent the night, centrifugal, the washing precipitation successively of ethanol, dehydrated alcohol, acetone, drying must be made with extra care Herb Gynostemmae Pentaphylli polysaccharides.
Get refining Herb Gynostemmae Pentaphylli polysaccharides 0.5g, add about 100ml distilled water, through 0.45 μ m membrane filtration.Adopt Q SepharoseFast Flow post, sample on the AKTA passage earlier with 2 column volumes of distilled water eluting, adopts sodium chloride to carry out gradient elution behind the last sample then, flow velocity 1ml/min, and ultraviolet inspection 210nm detects, and collects polysaccharide part GPS-3.Then it is concentrated the back and go up Superdex 200 posts, with 0.15mol/L sodium chloride eluting, flow velocity 0.5ml/min, ultraviolet 210nm detects, and collects the polysaccharide part, and respectively through sephadex G-25 desalination, lyophilization promptly gets homogeneous polysaccharide GPS-3 again.
Embodiment 2: Herb Gynostemmae Pentaphylli polysaccharides is to the protective effect of neurocyte amyloid-beta damage
Get pregnant 13.5-14 days Wistar rat, the cervical vertebra dislocation is put to death.
Aseptic condition separates down, takes out the embryo, in sterile petri dish, separates down in anatomic microscope, obtains neurocyte.Place CO2 gas incubator, 37 ℃, 5% carbon dioxide saturated humidity is carried out grouping experiment after cultivating 48h.
Neurocyte is cultivated after 48 hours in 96 orifice plates, adds the Herb Gynostemmae Pentaphylli polysaccharides of 5mg/ml embodiment 1 preparation respectively, makes the final concentration of Herb Gynostemmae Pentaphylli polysaccharides be respectively 100 μ g/ml, 200 μ g/ml, 300 μ g/ml, 400 μ g/ml, establishes the normal control group.
Cultivate after 12 hours, every hole adds the amyloid-beta culture fluid, and the final concentration that makes amyloid-beta is 1 μ mol/L, and 37 ℃, 5% carbon dioxide saturated humidity was cultivated after 36 hours, measured.
The result shows, damage has obvious protective effect to Herb Gynostemmae Pentaphylli polysaccharides to the neurocyte amyloid-beta, and its protective effect increases with the increase of activity, can the antagonism amyloid-beta more than the 200 μ g/ml to the nerve cell damage effect.Amyloid-beta is the important substance basis of senile dementia morbidity, and Herb Gynostemmae Pentaphylli polysaccharides shows that to the protective effect of this damage Herb Gynostemmae Pentaphylli polysaccharides can be effective to the prevention and the treatment of senile dementia.
Embodiment 3: Herb Gynostemmae Pentaphylli polysaccharides is to the effect of scopolamine induced mice dysmnesia model
1, experiment material:
1.1 experiment medicine:
Herb Gynostemmae Pentaphylli polysaccharides: prepare by example 1 method
Piracetam: 0.4g/ sheet, Tianjin Jinshi Pharmaceutical Co., Ltd..
Scopolamine hydrobromide injection: Shanghai Hefeng Pharmaceutical Co., Ltd.
1.2 laboratory animal: male mice in kunming, body weight (20 ± 2) g, available from Shandong University's Experimental Animal Center, the quality certification number: SCXK (Shandong) 20030004.
1.3 reagent and instrument:
Glutathion peroxidase (GSH-PX) test kit, superoxide dismutase (SOD) test kit, malonaldehyde (MDA) test kit, protein reagent box all build up bio-engineering research institute available from Nanjing.YYE50 mice diving tower instrument, instrument plant of Beijing Medical University produces.
2 models and grouping administration
2.1 group technology: experiment is divided into 6 groups, normal group, model group, piracetam group (0.2g/kg), administration group (0.1g/kg, 0.2g/kg, 0.4g/kg), every group of 10 mices.
2.2 medication: Herb Gynostemmae Pentaphylli polysaccharides, piracetam are all used physiological saline solution, every morning gastric infusion, normal group and model group are only given normal saline, continuous irrigation stomach 20 days, behind the last administration 1h, except that normal group, other each group group subcutaneous injection scopolamine 3.0mg/kg, the isopyknic distilled water of matched group subcutaneous injection.Behind the 10min, adopt the diving tower method to measure the mouse memory achievement.Behind the diving tower EOT, get cerebral tissue rapidly, weigh, make 10% homogenate with normal saline under the ice bath.Press the test kit explanation and measure GSH-PX, SOD vigor and MDA content.
3 experimental results
Table 1 is respectively organized the comparison (x ± s) of mice diving tower errors number
Annotate:
*Compare P<0.05 with model group,
*Compare P<0.01 with model group
As can be seen, model group mice diving tower errors number compares with model group apparently higher than matched group from last table data, and Herb Gynostemmae Pentaphylli polysaccharides group mice diving tower errors number all obviously reduces, and the doses effect relation is arranged.Illustrate that Herb Gynostemmae Pentaphylli polysaccharides can significantly improve the memory dysfunction that scopolamine causes.
Table 2 Herb Gynostemmae Pentaphylli polysaccharides is to the influence of mouse brain tissue GSH-PX, SOD activity and MDA content (x ± s)
Annotate:
*Compare P<0.05 with model group,
*Compare P<0.01 with model group
As can be seen, all than the remarkable rising of model group, MDA content significantly is lower than model group for GSH-PX, the SOD activity of each group of Herb Gynostemmae Pentaphylli polysaccharides, and presents dose-dependence from last table data.Illustrate that Herb Gynostemmae Pentaphylli polysaccharides can lower MDA content in the mouse brain, improve GSH-PX, SOD activity, have the effect of improving scopolamine learning memory disorder mouse memory ability.
Embodiment 4: the preparation of Herb Gynostemmae Pentaphylli polysaccharides tablet
Mainly form by Herb Gynostemmae Pentaphylli polysaccharides and adjuvant, wherein adjuvant can be in filler, disintegrating agent, binding agent, the lubricant partly or entirely, the consumption of adjuvant is no more than 8 times of Herb Gynostemmae Pentaphylli polysaccharides weight.
Preferred film-making prescription: in weight portion,
100 parts of Herb Gynostemmae Pentaphylli polysaccharides
100 parts of filleies
Speed collapses 90 parts of agent
80 parts of binding agents
6 parts of lubricants
Wherein: filler can be calcium hydrogen phosphate, dextrin or lactose; It can be L-HPC or dried starch that speed collapses agent; Binding agent can be PVP solution or starch slurry; Lubricant is a magnesium stearate.
The production technology of tablet is: press recipe quantity, take by weighing Herb Gynostemmae Pentaphylli polysaccharides, adding filler, speed collapse agent, mix homogeneously, the binding agent that adds recipe quantity then is mixed and made into wet grain, low temperature 40-65 ℃ drying, and it is even to add mix lubricant again, tabletting is used macromolecule dress material (as HPMC, EC, MC) bag film-coat at last.
Claims (8)
1. the application of Herb Gynostemmae Pentaphylli polysaccharides in preparation control medicine for senile dementia or health product.
2. with the Herb Gynostemmae Pentaphylli polysaccharides application of compositions in preparation control medicine for senile dementia or health product that active component is made.
3. application according to claim 1 and 2 is characterized in that: Herb Gynostemmae Pentaphylli polysaccharides or Herb Gynostemmae Pentaphylli polysaccharides compositions are made in injection, oral formulations, the external preparation any.
4. medicine that is used to prevent and treat senile dementia, it is characterized in that: its active component is a Herb Gynostemmae Pentaphylli polysaccharides, be aided with filler, disintegrating agent, binding agent and lubricant, wherein, the consumption of each component is as follows: 100 parts of Herb Gynostemmae Pentaphylli polysaccharides, 100 parts of filleies, speed collapse 90 parts of agent, 80 parts of binding agents, 6 parts of lubricants.
5. a kind of medicine that is used to prevent and treat senile dementia according to claim 4 is characterized in that: described filler is calcium hydrogen phosphate, dextrin or lactose.
6. a kind of medicine that is used to prevent and treat senile dementia according to claim 4 is characterized in that: it is L-HPC or dried starch that described speed collapses agent.
7. a kind of medicine that is used to prevent and treat senile dementia according to claim 4 is characterized in that: described binding agent is PVP solution or starch slurry.
8. a kind of medicine that is used to prevent and treat senile dementia according to claim 4 is characterized in that: described lubricant is a magnesium stearate.
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KR101811048B1 (en) * | 2016-09-12 | 2017-12-20 | 서울대학교산학협력단 | Composition comprising extract of Gynostemma longipes VK1 or longipenoside A compound isolated from thereof for preventing or treating of cognitive dysfunction |
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KR101811048B1 (en) * | 2016-09-12 | 2017-12-20 | 서울대학교산학협력단 | Composition comprising extract of Gynostemma longipes VK1 or longipenoside A compound isolated from thereof for preventing or treating of cognitive dysfunction |
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Open date: 20100127 |