CN106511492A - Preparation method for polygala japonica houtt extract and application for promoting hippocampal neurogenesis - Google Patents
Preparation method for polygala japonica houtt extract and application for promoting hippocampal neurogenesis Download PDFInfo
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- CN106511492A CN106511492A CN201611225855.XA CN201611225855A CN106511492A CN 106511492 A CN106511492 A CN 106511492A CN 201611225855 A CN201611225855 A CN 201611225855A CN 106511492 A CN106511492 A CN 106511492A
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- extract
- herba polygalae
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- polygalae japonicae
- polygala japonica
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- 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)
- A61K36/69—Polygalaceae (Milkwort family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention discloses a preparation method for polygala japonica houtt extract and application for promoting hippocampal neurogenesis. The preparation method comprises the steps of smashing, coarse extracting and ethanol precipitation. The prepared polygala japonica houtt extract is used for promoting hippocampal neurogenesis, experimental results show that hippocampal neural stem cell division can be improved by 50% through the polygala japonica houtt extract. The preparation process is easy to operate and very suitable for application and popularization; macromolecular compounds are effectively removed through the preparation method, and micromolecular amphoteric compounds are preserved to the greatest extent; the micromolecular amphoteric compounds can pass through a blood brain barrier and enter a brain, thus, compared with common water extracts, the effect of the polygala japonica houtt extract on promoting hippocampal neurogenesis is obvious, the polygala japonica houtt extract can be added into a drug carrier to form a mixture, the mixture is subjected to mixing, granulation and drying and then made into granules, capsules, tablets, oral liquid or injection according to a conventional drug preparation method; or the polygala japonica houtt extract is added into a carrier and made into health care beverages or health care food according to a conventional mehod.
Description
Technical field
The present invention relates to a kind of preparation method for promoting adult hippocampal cell proliferation of nerve cord complex, more particularly, to one
Plant and promote the value-added Herba Polygalae Japonicae method for preparing extractive of adult hippocampal neural stem cell.
Background technology
Hippocampus neural stem cells are located at hippocampal dentate granular cell lower region, and which can be in persistently division increasing in life
Grow.Neural stem cell division produces new neurocyte, migrates into hippocampal dentate area, and minority regenerative cell is converted into colloid
Cell, the overwhelming majority are converted into neurocyte, and after about 4-6 is all, neurocyte is ripe, are incorporated to existing neural circuit this mistake
Journey is referred to as nerve to occur.After growing up, hippocampal neural occurs safeguarding that brain plasticity has important effect.In adult, greatly
700 new neurocytes can be produced within about one day, 35% hippocampal dentate cell is replaced.Current study show that most of
The disease of nervous system is closed nerve to occur and changes related.Including depression, Alzheimer’s disease, parkinson disease and because declining
The diseases such as the cognitive competence decline that can cause always.Therefore, strengthen the disease that the adult nerve to occur of Hippocampus is potential nervous system to control
Reason method.
Herba Polygalae Japonicae, as a kind of Chinese herbal medicine with long history, with various biological function, is a kind of excellent rush
Enter the natural product that the adult neural stem cell of Hippocampus increases.Therefore there is huge potentiality to be exploited and using value.
The content of the invention
It is an object of the invention to plant Herba Polygalae Japonicae root as raw material, there is provided a kind of to promote adult hippocampal neural stem cell to increase
The Herba Polygalae Japonicae method for preparing extractive grown, is promoted the extract of nerve to occur effect, adds medicine using the extract
Or food carriers can prepare promotion nerve to occur affecting agent.
The method of the invention is comprised the following steps that：
（1）Crush：Herba Polygalae Japonicae root will be dried and be crushed to 20 ~ 50 mesh；
（2）It is thick to extract：The water of 10ml according to weight and volume ratio, is added in the above-mentioned Semen Benincasae bronzes of 1g, is filtered after boiling 2 hours,
The half of the 1st amount of water is added in obtaining filtrate, then slag after filtration, is filtered after boiling 2 hours；
（3）Ethanol precipitation：Collection step（2）The filtrate for obtaining, the concentration that 4 times of filtrates are added in filtrate are 100% second
Alcohol, 1-4 DEG C is stirred overnight, and supernatant is collected by centrifugation, and concentration, vacuum lyophilization obtain Herba Polygalae Japonicae extract.
Prepared by of the invention, complex is had an effect for promoting hippocampal neural：
The effect of this complex actor is a lot, but its outstanding role is to promote hippocampal neural to have an effect, and Herba Polygalae Japonicae extract is molten
100 ug/ml solubility Semen Benincasae gold solutions are prepared in normal saline, in intraperitoneal injection of mice body, continuous 7 days, daily 1ml；Control
Mice injects 1ml normal saline daily, or Herba Polygalae Japonicae is soluble in water（100 mg/ml）, continuous 7 days feeding mices（5 ml/
My god）；The daily normal water of control mice, promotees the effect that hippocampal neural occurs, at the 6th day, abdomen to count Herba Polygalae Japonicae extract
Inject 5- bromodeoxyuridine nucleoside in chamber（BrdU）.BrdU can replace thymus pyrimidine in cell propagation period（T）Penetrate into
The DNA molecular of duplication, then using anti-Brdu antibody, immunostaining shows proliferative cell；Take brain within 7th day, cut into slices, using exempting from
Hippocampal dentate granular cell lower region BrdU positive cell number in epidemic disease group technical Analysis brain, to calculate Hippocampus god
Jing stem cell divisions row is into newborn neurocyte number；As a result show, Herba Polygalae Japonicae extract can improve 50% hippocampal neural and do
Cell division.
The invention has the beneficial effects as follows：Preparation is simple, is especially suitable for popularization and application.In preparation method effectively
Remove macromolecular compound, as far as possible save small molecule amphoteric compound, these small molecule amphoteric compounds can pass through blood brain
Barrier, into brain.Therefore relative to common water extract, it is obvious that the extract promotees nerve to occur effect.Herba Polygalae Japonicae extract
Pharmaceutical carrier can be added to, be mixed, granulation is dried, then according to the medicament preparation method of routine makes granule or capsule
Agent or tablet or oral liquid or injection；Or carrier is added, conventionally make health beverage or health food.
Description of the drawings
Fig. 1 increased hippocampal dentate granular cell lower region for Herba Polygalae Japonicae；
In figure：Arrow is BrdU positive cells, and A, C are control mice, and B, D are Herba Polygalae Japonicae feeding mice, and arrow is that BrdU is positive
Cell；
Fig. 2 hippocampal dentate granular cell lower region BrdU positive cell number statistical results；
In figure：PJ is Herba Polygalae Japonicae, and control is to compare Mus, * * * P<0.01.
Specific embodiment
The preparation method of the Herba Polygalae Japonicae extract
1）To crush after Herba Polygalae Japonicae root clean dry, cross 20 mesh sieves, weigh 100g Semen Benincasae bronzes, add 1000mL water, boil 2h,
It is filtrated to get filtrate；Add 500ml water again in slag after filtration, boil extraction 2h, be filtrated to get filtrate；
2）Filtrate twice is collected, the ethanol of 4 times of filtrates is added, 4 DEG C are stirred overnight, filtrate is filtered to take in centrifugation, cold after concentration
Freeze vacuum drying, obtain final product rush nerve to occur effect Herba Polygalae Japonicae extract.
The application example of Herba Polygalae Japonicae extract：
1）Herba Polygalae Japonicae is dissolved in into normal saline 100ug/ml solution, daily lumbar injection 1ml Semen Benincasae gold solutions, continuous 7 days,
The daily injecting normal saline 1ml of control mice；Brdu was injected at the 6th day（50mg/kg）, the 7th day, brain is taken, cut into slices, PBS solution
Wash 3 times, 5 minutes every time, add Brdu mono- to resist 4 DEG C of overnight incubations, remove one and resist, PBS solution is washed 3 times, 5 minutes every time, is added
Two resist, and after one hour of incubation, remove two and resist, and PBS solution is washed 3 times, 5 minutes every time, adds Dapi（Nucleus dyestuff）Incubation 5
Minute；PBS solution washes 3 times, 5 minutes every time, mounting, in fluorescence microscopy Microscopic observation hippocampal dentate granular cell lower region
BrdU positive cell numbers
2）Herba Polygalae Japonicae is dissolved in into normal saline 100mg/ml solution.Feeding 5ml Semen Benincasae gold solutions, continuous 7 days, compare daily
The daily fed with water of mice, injected Brdu at the 6th day（50mg/kg）, the 7th day, brain being taken, cut into slices, PBS solution washes 3 times, every time 5 points
Clock, adds Brdu mono- to resist 4 DEG C of overnight incubations；Remove one to resist, PBS solution washes 3 times, every time 5 minutes；Add two to resist, be incubated one
After hour, remove two and resist, PBS solution is washed 3 times, 5 minutes every time, adds Dapi（Nucleus dyestuff）Incubation 5 minutes；PBS solution
Wash 3 times, 5 minutes every time, mounting, in fluorescence microscopy Microscopic observation hippocampal dentate granular cell lower region BrdU positive cell
Number, Fig. 1 are the hippocampal dentate granular cell lower floor fluorescence photo for shooting, and the fluorecyte of arrow indication is BrdU cells,
Represent the cell that new division is produced；Fig. 2 is hippocampal dentate granular cell lower region BrdU positive cell number statistical result,
As a result normal control Mus hippocampal dentate granular cell lower region BrdU positive cell number about 2200, Semen Benincasae are shown
Golden extract-treated hippocampus of mice dentate granule cell lower region BrdU positive cell number about 3500, it is newborn
Cell number increased 59%.
Claims (2)
1. a kind of preparation method of Herba Polygalae Japonicae extract, it is characterised in that methods described is comprised the following steps that：
（1）Crush：Herba Polygalae Japonicae root will be dried and be crushed to 20 ~ 50 mesh；
（2）It is thick to extract：The water of 10ml according to weight and volume ratio, is added in the above-mentioned Semen Benincasae bronzes of 1g, is filtered after boiling 2 hours,
The half of the 1st amount of water is added in obtaining filtrate, then slag after filtration, is filtered after boiling 2 hours；
（3）Ethanol precipitation：Collection step（2）The filtrate for obtaining, the concentration that 4 times of filtrates are added in filtrate are 100% second
Alcohol, 1-4 DEG C is stirred overnight, and supernatant is collected by centrifugation, and concentration, vacuum lyophilization obtain Herba Polygalae Japonicae extract.
2. a kind of Herba Polygalae Japonicae extract promotees the application that hippocampal neural occurs, it is characterised in that：
Herba Polygalae Japonicae extract prepares 100 ug/ml solubility Semen Benincasae gold solutions in being dissolved in normal saline, in intraperitoneal injection of mice body, even
It is continuous 7 days, daily 1ml；Control mice injects 1ml normal saline daily；Or Herba Polygalae Japonicae is soluble in water, Jia 100 in every ml water
Mg Herba Polygalae Japonicae, continuous 7 days feeding mices, daily 5 ml；The daily normal water of control mice；In order to count the increasing of Herba Polygalae Japonicae extract
The effect that strong hippocampal neural occurs, at the 6th day, lumbar injection 5- bromodeoxyuridine nucleoside was BrdU；BrdU can be in cell
Propagation period replaces thymus pyrimidine to penetrate into the DNA molecular for replicating, and then using anti-Brdu antibody, immunostaining shows and increases
Cell colonization；Take brain within 7th day, cut into slices, using hippocampal dentate granular cell lower region in immunohistochemistry technique analysis brain
BrdU positive cell numbers, go into newborn neurocyte number to calculate hippocampus neural stem cells division, as a result show, Semen Benincasae
Golden extract can improve 50% hippocampus neural stem cells division.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611225855.XA CN106511492B (en) | 2016-12-27 | 2016-12-27 | Preparation of Japanese polygala extract and application of Japanese polygala extract in promoting hippocampal neurogenesis |
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| CN201611225855.XA CN106511492B (en) | 2016-12-27 | 2016-12-27 | Preparation of Japanese polygala extract and application of Japanese polygala extract in promoting hippocampal neurogenesis |
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| CN106511492A true CN106511492A (en) | 2017-03-22 |
| CN106511492B CN106511492B (en) | 2019-12-20 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115212252A (en) * | 2022-08-18 | 2022-10-21 | 云南省中医医院(云南中医药大学第一附属医院) | Application of Va drug Nierone in preparation of drugs for treating cerebral infarction |
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| CN1651453A (en) * | 2004-12-09 | 2005-08-10 | 中国人民解放军第二军医大学 | Japanese polygala saponin kind compound and aglucon, total saponin and total aglucon and its application in medicine |
| CN1709903A (en) * | 2005-06-17 | 2005-12-21 | 中国人民解放军第二军医大学 | Saponin compound and use of its glucoside in preparing medicine for treating neure injure |
| CN101250508A (en) * | 2007-02-23 | 2008-08-27 | 株式会社秋田屋本店 | Queen gel clastic enzyme composition |
| CN102526550A (en) * | 2012-03-20 | 2012-07-04 | 江西南昌济生制药厂 | Preparation method for traditional Chinese medicine for resisting cyclomastopathy |
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| CN1709903A (en) * | 2005-06-17 | 2005-12-21 | 中国人民解放军第二军医大学 | Saponin compound and use of its glucoside in preparing medicine for treating neure injure |
| CN101250508A (en) * | 2007-02-23 | 2008-08-27 | 株式会社秋田屋本店 | Queen gel clastic enzyme composition |
| CN102526550A (en) * | 2012-03-20 | 2012-07-04 | 江西南昌济生制药厂 | Preparation method for traditional Chinese medicine for resisting cyclomastopathy |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115212252A (en) * | 2022-08-18 | 2022-10-21 | 云南省中医医院(云南中医药大学第一附属医院) | Application of Va drug Nierone in preparation of drugs for treating cerebral infarction |
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