CN107773580A

CN107773580A - For preventing or treating the crude drug composition of dementia or nerve degenerative diseases

Info

Publication number: CN107773580A
Application number: CN201710660953.4A
Authority: CN
Inventors: 谢心范; 林煌权; 山本理
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-08-25
Filing date: 2017-08-04
Publication date: 2018-03-09
Anticipated expiration: 2037-08-04
Also published as: CN107773580B; JP7005825B2; JP2019011305A

Abstract

The present invention, which provides, a kind of to be used to prevent or treats dementia or the crude drug composition of nerve degenerative diseases.The present invention relates to it is a kind of as effective crude drug composition contain pseudo-ginseng and the bark of eucommia, for preventing or treating dementia or its behavior and the composition of mental symptoms or nerve degenerative diseases.The invention further relates to it is a kind of as effective crude drug composition contain pseudo-ginseng, the bark of eucommia and sealwort, for preventing or treating dementia or its behavior and the composition of mental symptoms or nerve degenerative diseases.The invention further relates to it is a kind of as effective crude drug composition contain pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae, for preventing or treating dementia or its behavior and the composition of mental symptoms or nerve degenerative diseases.

Description

For preventing or treating the crude drug composition of dementia or nerve degenerative diseases

Technical field

It is used to prevent or treats dementia or the crude drug composition of nerve degenerative diseases the present invention relates to a kind of.More specifically Ground says, the present invention relates to it is a kind of as effective crude drug composition contain pseudo-ginseng and the bark of eucommia, for preventing or treating dementia or god Crude drug composition through degenerative disease.In addition, the present invention relates to one kind as effective crude drug composition contain pseudo-ginseng, the bark of eucommia and Sealwort, composition for preventing or treating dementia or nerve degenerative diseases.Moreover, it relates to a kind of be used as has Effect crude drug composition contain pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae, for preventing or treating dementia or nerve degenerative diseases Composition.

Background technology

Dementia refers to that the brain function of normal activity declines, and the cognitive function such as memory, thinking and judgment is gradually reduced, Hinder the state of daily life and social life.In modern society, 1 people just suffers from every 4 people in the crowd of more than 85 years old Dementia, the age is more big more easily to suffer from dementia, and illness rate of the over-65s less than 70 years old is 1.5%, has reached 27% within 85 years old, It is estimated that the Dementia patients of Japanese over-65s have been over 2,400,000 people, (MHLW " pays close attention to everybody mental health (body んなメ Application タ Le ヘ Le ス) " general website http://www.mhlw.go.jp/kokoro/know/disease_ recog.html).In recent years, dementia has become the problem of more and more important from now in aged society.

As shown in the table according to the investigation of police agency, it is 1,660,000 people to receive the people of cognitive function inspection in 2016, in advance Person under inspection will also continue to increase meter from now on.From the point of view of the result of cognitive function inspection during this is 8 years, the result of announcement is the 1st class (note Recall power and judgment reduction person) it is that the 2.3%, the 2nd class (memory and judgment slightly reduction person) is (memory of the 26.7%, the 3rd class Power and judgment do not have no problem person completely) (specify the homepage of federation of association of driving school in general civic organization all Japan for 71.0% " data of accident etc. " " being changed by inspection number for cognitive function inspection " http://www.zensiren.or.jp/kourei/ data/data.html)。

【Table 1】

Receive the number (in June, 2009~2016 year) of cognitive function inspection

General civic organization specifies federation of association of driving school to be compiled according to driving license section of Department of Communications of police agency data in all Japan Write

Dementia can be roughly divided into nerve degenerative diseases, cranial vascular disease in terms of the Basic disease cause of disease And other reasonses (including mental illness, Neurosurgical disease or internal disease).

So-called nerve degenerative diseases refer to cranial nerve cell slowly dead disease, cause by nerve degenerative diseases Dementia in, such as there is alzheimer dementia disease, Lewy body type dementia, cerebrovascular dementia, volume temporo type to be crazy about Slow-witted disease.

Patient's number of alzheimer dementia disease is most, occupies in dementia only about half of.Its pathogenic factor is recognized For the accumulation for being amyloid-beta in intracerebral and thus caused neurotoxicity, it was observed that the general atrophy of brain, brain The Histopathological Features such as room expansion, neurofibrillar change, senile plaque expelling, under decrease of memory, disorientation, judgment Drop etc. gradually aggravates, and some is also with mental symptoms such as depression.

What patient's number was only second to alzheimer dementia disease is Lewy body type dementia.Lewy body refers in nerve The specific proteins formed in cell, if research thinks that it is concentrated in the cerebral cortex or brain stem of brain, nerve cell It is destroyed and reduces, therefore can not successfully carry out nerve conduction, so as to triggers the symptom of dementia.

Equally, what patient's number was more also has cerebrovascular dementia, and this is due to cerebral infarction, cerebral hemorrhage, cerebral arteriovenous malformation It can not be delivered in nerve cell Deng nutrition and oxygen, the nerve cell death of its result part, neutral net receives destruction.

Volume temporo type dementia is also known as a creutzfeldt jakob disease, it is seen that frontal lobe and temporal atrophy, is one kind of juvenile dementia.

In addition, classifying in terms of symptom to dementia, core symptom and behavior and psychological disease can be roughly divided into Two kinds of shape (BPSD).

Core symptom is due to the symptom caused by the immediate cause i.e. destruction of brain cell of dementia, is typically exhibited Symptom have memory disorders, disorientation, understanding and judgment obstacle, perform function obstacle, the change of expression of feeling, aphasia, Agnosia, appraxia.

On the other hand, behavior and mental symptoms are also referred to as peripheral symptoms, also known as dementia patients behavior and psychosis in the past Shape (BPSD：Behavioral and Psychological Symptoms of Dementia).As behavior and mental symptoms Concrete example, such as hover, dietary behavior is abnormal, resist, anxiety and impatient, illusion and vain hope (stolen vain hope etc.), illusion, Depressed, excited, violence or abuse, incontinence, insomnia or sleep-disorder or reverse round the clock, requirement are gone home, eating disorder is with swallowing barrier Hinder, unclean behavior etc..

Nerve degenerative diseases are in addition to the Basic disease cause of above-mentioned dementia, and specific nerve is thin in also a kind of nervous centralis The gradual dead cranial nerve disease of born of the same parents group, for example, the op parkinson's such as amyotrophic lateral sclerosis (ALS), Parkinson's integrate Alzheimer dementias such as disease, senile agnosia etc., Progressive supranuclear palsy (PSP), Huntington disease, striatum substantia nigra It is denatured the polyphylys such as disease (SND), Xia Yi-De Leige syndromes (Shy-Drager syndromes), olive bridge cerebral atrophy (OPCA) The spinal cerebellar degeneration diseases (SCD) such as atrophy of uniting (MSA), spinocerebellar ataxia (SCA3), Freed deficiency disorder.

Brain forms nerve cell network, wherein the least unit for carrying out information transmission is nerve cell (neuron).God There is nervous process in order to form cerebral nervous system network through cell.Nervous process, which is morphologically generally divided into, sends information The projection (dendron) of projection (aixs cylinder) and receive information.Longer aixs cylinder and intricately branch are derived by a nerve cell Multiple projections of dendron, these projections are connected with other nerve cells, form the neutral net of complexity.

With the increase at age atrophy gradually occurs for brain, and it is difficult regeneration that damage, which once occurs, for cranial nerve cell, it may be said that Cell division once will not also occur after birth, almost identical cell can continue to use all one's life.Cranial nerve cell is before 5 years old Mushroom out, young brain can distally extend its branch with the growth of nerve cell, dendron, so as to which neutral net obtains Development.Once adult, the weight and volume reducing of brain, the quantity of cranial nerve cell are reduced with the increase at age.Especially It is in the dementia including alzheimer dementia disease or nerve degenerative diseases, it may occur that the atrophy of nervous process is with dashing forward Chu Minus are few, and these will cause memory disorders etc..

Therefore can expect, by recovering the atrophy of nervous process and reducing to improve the effect of dementia for cynapse.It is silly The cause of disease of slow-witted disease or even nerve degenerative diseases is varied, but is nerve net due to causing the immediate cause of neurological dysfunction The destruction of network, so if can make Neurite Outgrowth and rebuild neutral net, this will be to dementia or neurological Property disease prevention or treatment it is highly useful.

Nerve growth factor is also known as NGF (nerve growth factor), be represent maintain nerve cell survival, Promote one of neurotrophic factors of effect such as the growth of nervous process, the synthesis for promoting neurotransmitter, be by 3 subunits The protein complex that (α, β, γ) is formed.NGF β subunits be by 118 Amino acid profiles polypeptide (molecular weight 13kD), with The form of dimer is present.NGF is dispersed in peripheral nervous system in stomodaeal nervous system and sensory nerve system, also in maincenter In nervous system special role is played to being projected in cerebral cortex with the basal forebrain cholinergic nerve cell on hippocampus.Many institute's weeks Know, in the brain of Alzheimer disease severe detachment occurs for the neural cell group of NGF effects.Can therefore, it is desirable to probe into Improve growth Ahl tribulus sea silent sickness etc. due to the age and the research and development of the intracerebral NGF of reduction quantity or its active method with Mechanism.

On the other hand, it is well known that NGF it is related to inflammation, pain and obstacle (Freund V, Pons F, Joly V, Mathieu E,Martinet N,Frossard N.(2002)Upregulation of nerve growth factor Expression by human airway smooth muscle cells in inflammatory conditions. ( The up-regulation of expression of nerve growth caused by human body airway smooth muscle cells under inflammatory conditions) Eur Respir are J.20:458- 463).In addition, NGF activation mast cell, release histamine, this may help to treat histamine allergy (Aloe L, Skaper SD,Leon A,Levi-Montalcini R.(1994)Nerve growth factor and autoimmune Diseases.Autoimmunity. (nerve growth factor and autoimmune disease, autoimmunity) 19:141-150).Separately Outside, NGF and BDNF (Brain-derived neurotrophic factor；BDNF) it is identical, stimulate swollen The diffusion and survival of oncocyte, and promote new vessels in tumour formation (ChopinV, LagadecC, ToillonRA, LeBourhisX. (2016) Neurotrophin signaling in cancer stem cells. (god in cancer stem cell Through nutrition factor signal) Cell Mol Life Sci.73:1859-1870).So, the increase of NGF quantity be able to may not be brought Preferable result.

If it is possible to not increase NGF quantity too much and improve its nerve growth activity, this will be highly beneficial.

As the medicine of the Alzheimer disease of the typical example of dementia, the medicine developed earliest in the world is that he can Woods (trade name Cognex), got the Green Light in the U.S. within 1993, but because the damage to liver is larger, there is no in Japan and criticize It is accurate.Japan have been developed for it is small to hepar damnification, suppress decomposition in nerve cell in neurotransmitter acetylcholine, help In the medicine to regain the strength of memory, i.e., as acetylcholine degraded enzyme inhibitor donepezil (Aricept (registration mark), Eisai).Then, the medicine such as Rivastigmine, galanthamine, Memantine also obtain approval.

But the range of choice of the therapeutic agent for alzheimer's disease based on western medicine is still extremely limited, in addition, many Well known, medicine can also cause the side effect such as Spirit nerve symptoms of disease and symptom of digestive tract such as excited, spiritual agitation.The opposing party Face, in view of the limit and the misgivings of the side effect of conventional medication etc. to western medicine, in recent years, for dementia and its row For and mental symptoms, when have report to the validity of Chinese herbal medicine.As to the effective crude drug of dementia or Chinese herbal medicine, there is application Suppression liver dissipates, DANGGUI SHAOYAO SAN, suppression liver dissipate plus the example of tangerine peel, pinellia ternate etc..

For example, in recent years, the new possibility scattered as suppression liver is spiritual with neural field applied to dementia etc.. Crude drug Radix Angelicae Sinensis that suppression liver contains in dissipating, rhizome of chuanxiong, uncariae uncis cure ramulus are had been reported that if long due to easily causing the characteristic of gasteremphraxis Phase, which takes the breaking up of suppression liver, easily increases the burden of stomach and intestine.Therefore, addition dried orange peel and the tuber of pinellia in suppression liver dissipates, stomach is had concurrently by input The suppression liver of intestines protective effect dissipates plus tangerine peel, pinellia ternate, can significantly improve behavior and the mental symptoms of dementia, particularly aggressiveness It can be significantly improved (palace pool benevolence is bright, psychiatric department 14 (6), 535-542,2009).Add dried orange peel in addition, also having and suppression liver being dissipated The Clinical Report (rattan Tian how etc., psychiatric department 23 (1), 130-138,2013) of the tuber of pinellia.

On the other hand, as to triggering the diseases relevant with life style such as hypercholesterolemia, high blood pressure, diabetes Prevention and function improvement and hepatitis liver function the effective crude drug composition of improvement, containing pseudo-ginseng and the bark of eucommia Crude drug composition is widely known by the people with the crude drug composition containing pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae.

As the product containing pseudo-ginseng and the crude drug composition of the bark of eucommia, pseudo-ginseng bark of eucommia essence (registration mark) is widely known by the people, Can be from interpreter's industry Co., Ltd. of association (Tokyo) purchase.This is initially developed by the people in the present inventor, is contained There is the hot water extract using pseudo-ginseng and the bark of eucommia as the crude drug mixture of main component.The pseudo-ginseng bark of eucommia essence except main component pseudo-ginseng with And beyond the bark of eucommia, it can also preferably contain Korean ginseng, honey etc..

As the crude drug composition containing pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae, health Pien Tze Huang (registration mark) widely people Know, can be from interpreter's industry Co., Ltd. of association (Tokyo) purchase.This is also initially to be developed, contained by the people in the present inventor There is the crude drug composition of notoginseng extract, eucommia ulmoides extracts, Rhizoma Polygonati extract and licorice.It is in addition, hereinafter square Just for the sake of, the crude drug composition is described as " health Pien Tze Huang ".Crude drug composition containing pseudo-ginseng and the bark of eucommia and contain The crude drug composition of pseudo-ginseng, the bark of eucommia and sealwort (and radix glycyrrhizae), it is documented in Japanese Patent No. No. 4588132 specifications, the U.S. In No. 6,280,776 specification of patent and No. 6,586,017 specifications of U.S. Patent No. etc..

In addition, pseudo-ginseng bark of eucommia essence and health Pien Tze Huang are as healthy food application.

Look-ahead technique document Prior Art

Patent document

Patent document 1：Japanese documentation the 4588132nd

Patent document 2：American documentation literature the 6,280,776th

Patent document 3：American documentation literature the 6,586,017th

Non-patent literature

Non-patent literature 1：Greene LA and Tischler AS(1976)Proc Natl Acad Sci U S A, 73(7):2424-2428

Non-patent literature 2：Fukuda J,Yamaguchi K,Akimoto S,Tada Y.Neurosci Res.2:460- 471,1985

Non-patent literature 3：Fischer W,Wictorin K,Bjoerklund A,Williams LR,Varon S, Gage FH.Nature 329:65-68,1987

Non-patent literature 4：Freund V,Pons F,Joly V,Mathieu E,Martinet N,Frossard N. (2002)Upregulation of nerve growth factor expression by human airway smooth muscle cells in inflammatory conditions.Eur Respir J.20:458-463

Non-patent literature 5：Aloe L,Skaper SD,Leon A,Levi-Montalcini R.(1994)Nerve growth factor and autoimmune diseases.Autoimmunity.19:141-150

Non-patent literature 6：Chopin V,Lagadec C,Toillon RA,Le Bourhis X.(2016) Neurotrophin signaling in cancer stem cells.Cell Mol Life Sci.73:1859-1870

Non-patent literature 7：Palace pool benevolence is bright, psychiatric department 14 (6), 535-542,2009

Non-patent literature 8：Rattan Tian how grade, psychiatric department 23 (1), 130-138,2013

The content of the invention

Technical problem

In the prevention or treatment of dementia or nerve degenerative diseases, select to be adapted to the medicine of symptom and patient's constitution very Important, this will require a kind of to dementia, its behavior and mental symptoms or the highly effective medicine of nerve degenerative diseases. In this case, for by dementia or its related indication painful patient or the people around it, prevention method with The selection for the treatment of method should not only rely upon western medicine, east medical science should also enter with crude drug and traditional Chinese medicine field Row is expanded, it is therefore desirable to it is a kind of to dementia or its behavior and the prevention of mental symptoms (BPSD) or nerve degenerative diseases or Treat useful and less side effect crude drug composition.

In addition, nerve growth factor (NGF) shows the survival for maintaining nerve cell, the growth for promoting nervous process, promoted Enter the effect such as synthesis of neurotransmitter, but because the increase of NGF quantity may not can bring preferable result, therefore it is required that need not Increase NGF quantity and improve its nerve growth activity.

The present inventor etc. is carried out according to the activity of the nerve cells such as dementia or the relevant knowledge of crude drug composition Concentrate on studies, find a kind of have to dementia or its behavior and the prevention of mental symptoms or nerve degenerative diseases or treatment Crude drug composition, so as to complete the present invention.

Technical scheme

The present invention relates to it is a kind of as effective crude drug composition contain pseudo-ginseng and the bark of eucommia, for preventing or treating dementia Or its behavior and the composition of mental symptoms or nerve degenerative diseases.The invention further relates to one kind to be used as effective crude drug composition It is containing pseudo-ginseng, the bark of eucommia and sealwort, for preventing or treating dementia or its behavior and mental symptoms or nervus retrogression disease The composition of disease.The invention further relates to it is a kind of as effective crude drug composition contain pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae, be used for Prevention or treatment dementia or its behavior and the composition of mental symptoms or nerve degenerative diseases.

So far, the crude drug composition containing pseudo-ginseng and the bark of eucommia, the combination of the crude drug containing pseudo-ginseng, the bark of eucommia and sealwort Thing and crude drug composition containing pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae are to dementia or its behavior and mental symptoms or god Prevention or the useful this point for the treatment of through degenerative disease are completely unknown by the people in this area, even also without any enlightenment.

The present invention is as described below.

(1) a kind of crude drug composition, for preventing either to treat dementia or its behavior and mental symptoms or nerve Nerve degenerative disease, the crude drug composition contain the pseudo-ginseng and the bark of eucommia as crude drug composition.

(2) as claim 1 of the crude drug composition also containing sealwort described in crude drug composition.

(3) as above-mentioned (1) or (2) of the crude drug composition also containing radix glycyrrhizae described in crude drug composition,.

(4) crude drug composition described in above-mentioned (1), the pseudo-ginseng are 10~90 weight %, and the bark of eucommia is 10~90 weights Measure %.

(5) crude drug composition described in above-mentioned (2), the pseudo-ginseng are 10~90 weight %, and the bark of eucommia is 10~90 weights % is measured, the sealwort is 2~25 weight %, limits each crude drug component content sum as 100 weight %.

(6) crude drug composition described in above-mentioned (3), the pseudo-ginseng are 10~90 weight %, and the bark of eucommia is 10~90 weights % is measured, the sealwort is 2~25 weight %, and the radix glycyrrhizae is 3~20 weight %, limits each crude drug component content sum as 100 Weight %.

(7) crude drug composition described in above-mentioned (3), the pseudo-ginseng are 10~90 weight %, and the bark of eucommia is 10~90 weights % is measured, the sealwort is 2~25 weight %, and the radix glycyrrhizae is 5~20 weight %, limits each crude drug component content sum as 100 Weight %.

(8) crude drug composition described in any one of above-mentioned (1)~(7), the crude drug composition are each crude drug composition Extract.

(9) crude drug composition described in any one of above-mentioned (1)~(8), the dementia or its behavior and psychological disease The prevention or treatment of shape or nerve degenerative diseases are entered by the expression for increasing neurofilament or the growth for promoting nervous process OK.

Beneficial effect

It is dementia or its behavior and mental symptoms or nerve degenerative diseases by the crude drug composition of the present invention Prevention or treatment provide new selection.

The growth of expression or nervous process of the crude drug composition of the present invention to neurofilament is effective, particularly and if NGF It is used together, even not showing the NGF of the relatively low usage amount of effect in NGF itself, can also promotes the expression of neurofilament And the growth of nervous process.

Brief description of the drawings

Fig. 1 is propagation of smart (DTS) extract of the display pseudo-ginseng bark of eucommia with health Pien Tze Huang (YHK) extract to PC12 cells Effect chart；Cultivate cell PC12 cells DTS extracts and YHK extract-treateds 48 hours.Entered using MTT colorimetric methods Row cell survival and proliferation test.It is worth for average value ± SEM, takes 5 samples, n=4 respectively；

Fig. 2 for display be used alone DTS extracts and YHK extracts in PC12 cells whether the life of inducing neural projection Long micro-image and bar chart；

(A) DTS or YHK extracts (0.1,0.5 and 5mg/mL) 48 hours are applied on culture PC12 cells.Utilize The paraformaldehyde of iced storage 4% fixes cell.Bar shaped represents 50 μm.NGF is provided as positive control.Represent from 4 independent experiments Obtained representative image；

(B) culture PC12 cells are carried out and (A) same processing.The length of percentage and nervous process to noble cells Spend and counted described in the method paragraph according to aftermentioned embodiment.Value is expressed as 100 total cells hundred counted in cell Divide ratio.Mean+/-standard error, n=4；

The electrophoresis that YHK extracts induce the neurofilament in PC12 cells to express in high concentration is used alone for display in Fig. 3 Image and histogram；

(A) using DTS extracts or YHK extracts (0.1,0.5 and 5mg/mL) 48 hours on culture PC12 cells. Neurofilament (NF68 and NF160) expression is determined by specific antibody.Compareed as internal reference and GAPDH is provided.Represent from 4 solely It is vertical to test obtained representative image；

(B) quantitative result is represented with histogram.Value is represented with the multiple of a reference value (Basal).Control value is set as 1.It is flat Mean value ± standard error, n=4.Compare control value, * p ＜ 0.05, * * p ＜ 0.01 and * * * p ＜ 0.001；

Fig. 4 be display DTS extracts and YHK extracts promote micro-image by the Neurite Outgrowth of NGF inductions with Bar chart；

(A) by DTS extracts or YHK extracts (0.1,0.5 and 5mg/mL) and NGF on culture PC12 cells (0.5ng/mL) is applied 48 hours jointly.NGF (50ng/mL) has been used as positive control.Utilize the paraformaldehyde of iced storage 4% Fixed cell, it was observed that the growth of nervous process.Bar shaped represents 50 μm；

Fig. 5 be display DTS extracts and YHK extracts promote the electrophoretic image that the neurofilament by NGF inductions is expressed with it is straight Fang Tu；

(A) by DTS extracts or YHK extracts (0.1,0.5 and 5mg/mL) and NGF on culture PC12 cells (0.5ng/mL) is jointly processed by 48 hours.Neurofilament (NF68 and NF160) expression is determined by specific antibody.As internal reference Control provides GAPDH.Represent the representative image obtained from 4 independent experiments；

Fig. 6 is the table with together showing inspection result during taking according to Xing Do and age bracket；

Fig. 7 shows the content of the prediction test of big friendly formula dementia；

Fig. 8 is the chart of the analysis that to take year related of subjects of the display to only taking YHK；

Fig. 9 is the chart of the analysis that to take year related of subjects of the display to only taking DTS；

Figure 10 is display and the chart for the analysis that to take year related for taking YHK and both DTS subjects；

Figure 11 is the chart for the analysis for showing that the age of the subjects to only taking YHK is related；

Figure 12 is the chart for the analysis for showing that the age of the subjects to only taking DTS is related；

Figure 13 is the chart for showing the analysis related with the age of both DTS subjects to taking YHK.

Embodiment

Crude drug composition in the present invention is illustrated.

In the present invention, crude drug composition refers to the various crude drugs of composition crude drug composition, and the various crude drugs can be its Ji Yuan The medicinal effects of plant or its powder, can also be either obtained from its extract or can also be dries extract Particle etc., the granule being adjusted to it, tablet, pill, powder, capsule, solution (drink agent), extract, tincture, The form of the various formulations such as syrup, alcohol agent.

In the present invention, pseudo-ginseng be also known as pseudo-ginseng ginseng, Tienchi-Ginseng, Notoginseng, Radix Notoginseng, Or Pseudoginseng etc., it is defined as the original Araliaceae medicinal plant pseudo-ginseng ginseng Panax notoginseng's of China Root or Panax pseudo-ginseng Wall or Panax pseudo-ginseng Wall.var.notoginseng (Burkill) Hoo＆Tseng etc. root.It is well known that the Chinese medicine can improve Dyslipidemia, suppress hypertension and pain Bitterly.In the present invention, the Original plant as pseudo-ginseng, the plant used in this area as other Original plants can also be used Thing.In the present invention, pseudo-ginseng is not particularly limited using position, usually using its root.

The bark of eucommia is also known as Eucommia Bark, EUCOMMIAE CORTEX, generally by the high wooden bark of eucommia of Eucommiaceae fallen leaves Eucommiae ulmoides bark dried object composition.It is well known that the crude drug has enhancing liver function and renal function and dropped The effect of low hypertension and high fat of blood concentration.In the present invention, the Original plant as the bark of eucommia, can also use in the art The plant used as other Original plants.In the present invention, the bark of eucommia is not particularly limited using position, but in the present invention In, the bark of the Original plants such as the above-mentioned bark of eucommia is referred not only to, also refers to its young leaves (Eucommiae Folium), fruit (Eucommiae Berry) and/or trunk part (Eucommiae Wood) dried object.

Sealwort is also known as Polygonatum Rhizome, Polygonati Rhizoma or Siberian Solomonseal Rhizome, be to improve nutritional status, improve health useful traditional crude drug.It is well known that the crude drug pair Symptom and the liver function for improving chronic hepatitis patient are effective.Sealwort is not particularly limited using position, but in the present invention, Sealwort refers to that the rhizome of the Original plant to sealwort (Polygonatum) category is dried or carried out product obtained by boiling.

In the present invention, the Original plant as sealwort, yellow radix bupleuri (Polygonatum falcatum are used as representing A.Gray), polygonatum sibiricum Redoute (Polygonatum sibiricum Redoute, Polygonatum sibiricum Delar.ex Redoute)、Polygonatum kingianum Collett et Hemsley、Polygonatum cyrtonema Hua (Liliaceae) (Pharmacopeia of Japan is corrected with reference to the 17th).

In the present invention, the Original plant as sealwort, radix polygonati officinalis (Polygonatum can also be used as needed odoratum(Mill.)Druce、Polygonatum odoratum)、Polygonatum multiflorum、 Polygonatum stenophyllum Maxim.、Polygonatum involucratum Maxim.、Polygonatum macropodium Turez.、Polygonatum cirrhifolium(Wall.)Royle、Polygonatum prattii Baker、Polygonatum punctatum Royle ex Kunth、Polygonatum zanlanscianense Pamp.、 Polygonatum curvistylum Hua、Polygonatum tessellatum Wang et Tang、Polygonatum roseum(Ledeb.)Kunth、Polygonatum verticillatum(L.)All.、Polygonatum curvistylum Hua、Polygonatum erythrocarpum Hua、Polygonatum filipes Merr.、Polygonatum Lasianthum Maxim. etc..

In the present invention, radix glycyrrhizae is also known as glycyrrhiza, glycyrrhizae radix, Glycyrrhizae Rhizoma, licorice or liquorice, it is that relatively broad crude drug is used in Chinese herbal medicine.It is defined as the radix glycyrrhizae of pulse family (Glycyrrhiza) Original plant of category is the root, rhizome or the product that stolon is dried of hay class.

In the present invention, the Original plant as radix glycyrrhizae, correct what is included in Japanese officina using the 17th as representing Glycyrrhiza uralensis Fischer (Glycyrrhiza Uralensis) or Glycyrrhiza glabra Linne (Leguminosae) (Spanish licorice).

In the present invention, the Original plant as radix glycyrrhizae, Glycyrrhiza can also be used as needed Acanthocarpa, G.aspera, G.astragalina, G.bucharica, Russian radix glycyrrhizae G.echinata, G.eglandulosa、G.foetida、G.foetidissima、G.gontscharovii、G.iconica、 G.korshinskyi, U.S. radix glycyrrhizae G.lepidota, G.pallidiflora, G.squamulosa, G.triphylla, G.yunnanensis, Xinjiang radix glycyrrhizae G.inflata etc..

In the present invention, the Original plant as radix glycyrrhizae, can also use makes in the art as other Original plants Plant.In the present invention, radix glycyrrhizae is not particularly limited using position, but as described generally above using root, rhizome or Stolon.

In the crude drug composition of the present invention, the usage amount of pseudo-ginseng is 10~90 weight %, be preferably 30~80 weight %, More preferably 35~75 weight %, it is more preferably 40~60 weight %.Usage amount can insufficient above range most lower limit or More than the most upper limit, but due to the cooperative effect with other crude drug compositions can not be played sometimes, therefore it is preferably within the above range Use.

In the crude drug composition of the present invention, the usage amount of the bark of eucommia is 10~90 weight %, be preferably 15~80 weight %, More preferably 20~60 weight %, it is more preferably 30~50 weight %.Usage amount can insufficient above range most lower limit or More than the most upper limit, but due to the cooperative effect with other crude drug compositions can not be played sometimes, therefore it is preferably within the above range Use.

In the crude drug composition of the present invention, the usage amount of sealwort is 0~25 weight %, is preferably 2~25 weight %, more Preferably 6~20 weight %, it is more preferably 10~20 weight %.Usage amount can insufficient above range most lower limit or exceed The most upper limit, but due to the cooperative effect with other crude drug compositions can not be played sometimes, therefore be preferably to use within the above range. Particularly, it is difficult to be digested if the ratio of sealwort is excessive, therefore is not easy to be absorbed by the body, and causes to have a stomach upset, It is advantageous to use within the above range.

In the crude drug composition of the present invention, the usage amount of radix glycyrrhizae can be 0~20 weight %.In the crude drug group of the present invention In compound, in the case of using sealwort as crude drug composition, if it is considered that its cooperative effect, it is preferable to use radix glycyrrhizae.At this In the case of kind, the usage amount of radix glycyrrhizae can be preferably 3~20 weight %, more preferably 5~20 weight %, be more preferably 10~ 20 weight %.Usage amount can insufficient above range lower limit or more than the most upper limit, but due to can not play sometimes and other life The cooperative effect of medicine composition, therefore be preferably to use within the above range.In addition, on radix glycyrrhizae, it is contemplated that its excessive intake is drawn The side effect risen, is preferably used within the above range.Wherein, each crude drug component content sum is limited as 100 weight % (to work as Above-mentioned each crude drug composition is in its weight % span, it is necessary to meet that all crude drug compositions take the weight of content during value The summation for measuring % is 100 weight %).

On the composition of above-mentioned crude drug composition, as long as each crude drug composition in the range of above-mentioned composition, as described below, is gone back Other crude drug compositions can be contained.For example, the present invention crude drug composition as any crude drug composition can also contain be selected from by At least one of the group of Korean ginseng and honey composition.Wherein, it is 100 weight % (i.e. when upper to limit each crude drug component content sum State each crude drug composition in its weight % span during value, it is necessary to meet that all crude drug compositions take the weight of content The summation for measuring % is 100 weight %).

In the crude drug composition of the present invention, any crude drug composition is that the usage amount of Korean ginseng can be 0~20 weight %, In addition, the usage amount that any crude drug composition is honey can be 0~30 weight %.In view of the easy intake of crude drug composition, These compositions preferably use within the above range.

In addition, above-mentioned crude drug composition and any crude drug composition have in collection time, season, moisture of each raw material etc. Institute's difference, therefore above-mentioned usage amount can suitably be changed, it can also be used outside above range.

The crude drug composition of the present invention, by making it contain these optional members, have and be easy to intake and easily by people Body absorbs, it is short-term take it is i.e. effective, even if terminate take after its effect can also continue the advantages of.At this point, make During with sealwort, it is expected to make it contain radix glycyrrhizae for most Japaneses, but according to individual difference, even if there also have some not contain to be sweet The example that grass can also be absorbed by the body.

In the present invention, any crude drug composition can also use usually used crude drug.Any crude drug composition is if example Such as the kind that Pharmacopeia of Japan is included, can also use but it is also possible to be the kind beyond it.In addition, the life as the present invention Medicine composition, it can also use above-mentioned in the commodity pseudo-ginseng bark of eucommia that can be bought on the market essence and health Pien Tze Huang.

In the present invention, when mentioning each crude drug composition such as pseudo-ginseng, the bark of eucommia, sealwort, radix glycyrrhizae or crude drug composition, Original plant Medicinal effects, that is, during the medicinal effects of Original plant, can also mean that its root, stolon, bark, young leaves, Any one of fruit, trunk part, rhizome and/or their extract.In addition, its formulation can be powder, granule, piece Any one of agent, pill, powder, capsule, solution (drink agent), extract, tincture, syrup, alcohol agent etc..

In addition, in the present invention, " extract of crude drug composition " or " extract " refer to the i.e. crude drug composition of extracting object, In the water of room temperature~60 DEG C or 60 DEG C~100 DEG C of hot water extract or the ethanol of room temperature~100 DEG C or water and ethanol Mixed solution (100：0~0：100) extract.Extraction time can be such as 0.5 hour~2 hours, but according to its purpose The shorter or longer time can also suitably be used.Species, temperature and the time of said extracted solvent are example, according to mesh Can suitably change.

In addition, the crude drug composition in crude drug composition by extraction process is not carried out, the medicinal effects of Original plant, i.e., its When root, stolon, bark, young leaves, fruit, trunk part, rhizome etc. form and/or, when being the formulation containing them, can To implement to use after extracting as described above before use.

Crude drug is the total of the medicine that need not purify active ingredient from the product with naturally occurring drug effect and use Claim, in addition, active ingredient is also not limited to one kind, Multiple components are intricately interrelated in many cases.Therefore, present crude drug Composition or its extract are made up of which kind of active chemical or how being capable of the directly specific crude drugs by structure or characteristic Composition or its extract, this is by as problem from now on.

Crude drug composition (not containing sealwort) of the invention containing pseudo-ginseng and the bark of eucommia, except main crude drug composition pseudo-ginseng and Outside the bark of eucommia, as optional member it can also be made to contain Korean ginseng, honey etc..As the crude drug combination containing pseudo-ginseng and the bark of eucommia Thing (does not contain sealwort), as described above, pseudo-ginseng bark of eucommia essence (registration mark, interpreter's industry Co., Ltd. of association (Tokyo)) is widely known by the people, And on sale on the market.In the description of the present application, for convenience's sake, it will contain pseudo-ginseng and the bark of eucommia sometimes and not contain The crude drug composition of sealwort is referred to as DTS.

The crude drug composition containing pseudo-ginseng, the bark of eucommia and sealwort of the present invention, preferably makes it contain radix glycyrrhizae.As containing Pseudo-ginseng, the bark of eucommia, the crude drug composition of sealwort and radix glycyrrhizae, as described above, health Pien Tze Huang (registration mark, association's interpreter's industry strain formula Commercial firm (Tokyo)) it is widely known by the people, and on sale on the market.It contains using pseudo-ginseng, the bark of eucommia, sealwort and radix glycyrrhizae as main component Crude drug mixture hot water extract.In the description of the present application, for convenience's sake, sometimes will contain pseudo-ginseng, the bark of eucommia, Sealwort and the crude drug composition of radix glycyrrhizae are referred to as YHK.

Then, the manufacture method of the crude drug composition of the present invention is illustrated.

The crude drug composition of the present invention can use above-mentioned crude drug composition and be carried out according to method commonly used in the art Manufacture.

For example, crude drug composition in crude drug composition by be not carried out the root of extraction process, stolon, bark, young leaves, When fruit, trunk part, rhizome etc. form and/or, when being the formulation containing them, can be by the way that they be quantitatively weighed simultaneously Mixed, manufacture the crude drug composition of the present invention.

When using the crude drug composition that extraction process is not carried out, such as powder can be made and used.

The crude drug composition can be used with recipe quantity of subpackage 1 time etc..In this case, the life of the invention of prescription Drug composition, it can be taken after using preceding equally implementation extraction as described above.Resulting liquid extract can be by dry It is dry that particle is made, as described below, granule, tablet, pill, powder, capsule, solution (drink agent), leaching can be made The form of the various formulations such as paste, tincture, syrup, alcohol agent.

In addition, other aspects of the crude drug composition as the present invention, for example, each crude drug composition can be the crude drug composition Extract.Such as shred is made in above-mentioned each crude drug composition and is quantitatively weighed, for above-mentioned each crude drug composition for being weighed or The mixture that they are obtained by mixing by person, as described above, entering by using the mixed solution of water, hot water, ethanol or water and ethanol Row extraction, and as needed through filtering, purifying, each process of concentration, the crude drug composition of liquid extract can be manufactured.

Can be by the recipe quantity subpackage of 1 time in the hermetic bags such as aluminium bag.Now, the crude drug composition of the invention of prescription, Can be using preceding being heated or taken after being diluted.

In the present invention, the extracting method of crude drug composition is not limited to the above method, can use common according to this area The usual any extracting method of the general knowledge of technical staff.

Particle can be made by drying in the extract of resulting crude drug composition, as described below, particle can be made The shape of the various formulations such as agent, tablet, pill, powder, capsule, solution (drink agent), extract, tincture, syrup, alcohol agent Formula.

As desired as optional member, Korean ginseng extract, honey etc. can be added into two obtained extracts and entered Row mixing (mixed processes).By carrying out heated-air drying (drying process) to mixture, powdered or even powder can be obtained Granular crude drug composition of the invention.

In addition, being shaped by adding the excipient such as emulsifying agent into obtained mixture and carrying out heated-air drying, can obtain To tablets and other formulations.As emulsifying agent, honey, vegetable oil, xylitol etc. can be used.The weight ratio of emulsifying agent, preferably 0~ 20 weight %.

The mixed-powder of the extract of above-mentioned crude drug composition and as needed to emulsifying agent carry out heated-air drying and obtain Tablet in the content of crude drug composition of the invention be 0.1~0.5g/ tablets, particularly preferably 0.25g/ tablets.

In human body, from the viewpoint of validity, the intake of the crude drug composition of the present invention is not particularly limited, Set out from a security point, 0.1~2g/Kg on the about 1st, be preferably 0.1~0.25g/Kg.

But from the viewpoint of validity, security, according to the common technical knowledge of those of ordinary skill in the art, these The content and intake of crude drug composition, which may be set in, not to be produced side effect or is controlled in irreducible minimum and can be abundant Play the amount of drug effect.

The crude drug composition of the present invention, can be administered according to the form of administration using various formulations.In order to formulation, Diluent or the excipient such as filler, extender, adhesive, wetting agent, disintegrant, surfactant can generally be used.It is suitable Close be administered orally mode, as formulation have powder, granule, tablet, pill, powder, capsule, solution (drink agent), The formulations such as extract, tincture, syrup, alcohol agent, by using diluent and excipient, it can be obtained according to the form of these formulations To the crude drug composition of the present invention.The formulation of solid pharmaceutical preparation can at least use a kind of excipient, such as starch, calcium carbonate, sugarcane Sugar, lactose, gelatin etc., in addition, except simple excipient can also use the lubricant of magnesium stearate, talcum powder etc.Liquid The formulation of body preparation can use suspending agent, emulsifying agent, syrup etc., in addition, as simple diluent can also use water, Wetting agent, sweetener, aromatic, antistaling agent etc..

By formulation crude drug composition of the invention, subpackage and commercialization suitably can be carried out according to purpose.For example, can To carry out subpackage and commercialization with hermetic bags or subpackage paper etc. such as aluminium bag, aluminium foil, damp proof fibrous papers.

It is powder, granule, tablet, pill, powder, capsule, solution (drink agent), medicinal extract on crude drug composition The manufacture method of the crude drug composition of the invention of agent, tincture, syrup, alcohol agent or the like the form of other formulations, this area Those of ordinary skill can relatively easily understand manufacture method by crude drug and pharmaceutical industry general knowledge and described above.

Hereinafter, to crude drug composition and dementia or its behavior and mental symptoms or the nervus retrogression disease of the present invention Sick related pharmacological effect illustrates.

It is well known that PC12 cells are derived from the cell line of the pheochromocytoma by the adrenal medella derivation from rat (Greene LA and Tischler AS(1976)Proc Natl Acad Sci USA,73(7):2424-2428), if Nerve growth factor (NGF) is played a role, then Neurite Outgrowth and be divided into sympathetoblast shape.Therefore, PC12 is thin Cell in vitro of the born of the same parents as the differentiation mechanism in the growth for explaining nervous process, nerve cell, the mechanism of explaination NGF effects Model, it is used to study since very early, and in (Das KP, Freudenrich TM, Mundy WR. (2004) on sale on the market Assessment of PC12 cell differentiation and neurite growth:a comparison of The morphological and neurochemical measures. (assessments of PC12 cell differentiations and Neurite Outgrowth：Shape The comparison of state and neurochemical measure) Neurotoxicol Teratol.26:397-406).

NGF is dispersed in peripheral nervous system in stomodaeal nervous system and sensory nerve system, in addition, in central nervous system In system special role is played to being projected in cerebral cortex with the basal forebrain cholinergic nerve cell on hippocampus.It is well known that NGF In the brain of Alzheimer disease severe detachment occurs for the neural cell group of effect.Friendships of the NGF to Elderly patients and rat Sense ganglion neuronal also shows that extensional process (Fukuda J, Yamaguchi K, the Akimoto S, Tada of nerve fibre Y.NGF-dependent and-independent growth of neurites from sympathetic ganglion Cells of the aged human in a serum-free culture. be (the elderly's in serum free medium NGF- dependences and the dependent/non-dependent growth of the nervous process of sympathetic ganglionic cell) Neurosci Res.2:460-471, 1985).In addition, also imply that the decline (Fischer related to the decline of the learning ability caused by aging of intracerebral NGF activities W,Wictorin K,Bjoerklund A,Williams LR,Varon S,Gage FH.Amelioration of cholinergic neuron atrophy and spatial memory impairment in aged rats by Nerve growth factor. (the cholinergic neuron atrophy of aging rats caused by nerve growth factor and spatial memory barrier The improvement hindered) Nature329:65-68,1987).

Neurofilament is the special median fiber in nerve cell, it is sparse in neurocyte and dendron and in aixs cylinder Exist interior comparatively dense, be connected with each other by countless crosslinkings.It has been distinguished that, intrinsic albumen is different by molecular weight in neurofilament 3 kinds of homology isomer compositions, molecular weight 200kD referred to as NF-H, molecular weight 160kD referred to as NF-M or NF160, molecule Measure 68kD referred to as NF-L or NF68.

The expression of neurofilament (NF) Protein high between axon growth is controlled to imply that hair of the neurofilament to aixs cylinder Open up particularly significant.According to some researchs, imply that neurofilament is by long Neurite elongation, can more efficiently promote in aixs cylinder The extension (Walker KL, Yoo HK, Undamatla J, SzaroBG. (2001) Loss of neurofilaments (loss of neurofilament changes axon growth dynamic to alters axonal growth dynamics..)J Neurosci.21: 9655-9666)。

It may determine that, crude drug composition of the invention, the expression of inducing neural silk in PC12 cells, therefore in PC12 Trigger the growth of nervous process that there is significant effect in cell.If in addition, the crude drug composition of the present invention makes together with NGF With the expression of the neurofilament in the further PC12 cells promoted by NGF inductions.Furthermore if crude drug composition of the invention It is used together with NGF, is obviously promoted the growth of the nervous process by NGF inductions.

It may determine that, the dementia or its behavior and the prevention or treatment of mental symptoms or nerve degenerative diseases, lead to Cross the expression for the neurofilament that increase is induced by nerve growth factor (NGF) and/or promote the life of the nervous process in nerve cell Grow and carry out.Therefore, crude drug composition of the invention, to the expression containing neurofilament and the growth of the nervous process of nerve cell , the differentiation of nerve cell has significant effect with growth, therefore dementia or its behavior and mental symptoms or nerve moved back The prevention or treatment of row disease have good effect.

It is related to the effect of the invention of dementia or its behavior and the prevention of mental symptoms or nerve degenerative diseases or treatment Fruit, it can also be verified by answering application form to the subjects that DTS or YHK has been taken within certain period.

Big friendly formula dementia prediction test is employed in the application form for simply checking dementia (to take passages certainly Public good juridical person dementia prevents the homepage http of financial group://www.mainichi.co.jp/ninchishou/ explanation.html).Its content is shown in Fig. 6.This test is designed as the elderly in itself or spouse person or cohabitant etc. The pole early stage of dementia, the early stage of dementia or the state that may develop to dementia can simply be predicted.

From the result of application form, take both YHK, DTS or YHK of the present invention and DTS, the prevention of dementia and Treatment is effective.

Hereinafter, by embodiment, the present invention will be described in more detail, but these embodiments are only example, and does not have to To limit the scope of the present invention determined by following claims.

Embodiment

Embodiment 1

[purpose]

In order to probe into the function of pseudo-ginseng bark of eucommia essence and health Pien Tze Huang to nerve cell, pseudo-ginseng bark of eucommia essence and health are prepared for The water extract of Pien Tze Huang, the effect of pseudo-ginseng bark of eucommia essence and health Pien Tze Huang is assessed according to neuropathology.Pseudo-ginseng The neurobiological function of bark of eucommia essence and health Pien Tze Huang from the differentiation of (i) nerve cell and the expression of (ii) synapsin this Two aspects are assessed.

[materials and methods]

(1) preparation of crude drug composition (extract)

Used pseudo-ginseng bark of eucommia essence is from interpreter's industry Co., Ltd. of association (Tokyo) purchase, and it is by pseudo-ginseng 33g (55 weight %) And bark of eucommia 22g (45 weight %) compositions.Hereinafter, the crude drug composition is referred to as DTS.

Used health Pien Tze Huang is by pseudo-ginseng 6g (40 weight %), bark of eucommia 4.5g (30 weight %), sealwort 2.25g (15 Weight %) and radix glycyrrhizae 2.25g (15 weight %) compositions.Hereinafter, the crude drug composition is referred to as YHK.

DTS and YHK each sample is ground into powder, 50mg is weighed into the micro-pipe of 1.5mL Ai Bende companies, adds Enter 1mL water, carry out 30 minutes ultrasonic irradiations, centrifuged 10 minutes with 13200rpm.Supernatant is collected, is noted by PTFE (0.22 μm) filtering of discharging type filter.Stoste is 50mg/mL, is preserved at -20 DEG C.

(2) cell culture

PC12 cells from rat adrenal medulla are from American Type Tissue Culture (American Type Culture Collection, ATCC) purchase.With the hyclone (FBS) and horse serum (HS), 100 units that with the addition of 6%/ Dulbecco's modified Eagle medium (DMEM) of mL penicillin and 100 μ g/mL streptomysins is trained to cell Support, be positioned at 37 DEG C in CO2 (7.5%) incubator (incubator) of humidification.New culture medium is applied daily.Culture FBS and HS of the base before 3 hours of extraction process with 1%, 100 units/MI penicillin and 100 μ g/mL streptomysin one Reinstate new DMEM displacements.In extraction process, with containing 1% FBS and HS, 100 units/mL penicillin and 100 μ g/ The DMEM of mL streptomysin dilutes extract.

(3) cell survival assay

The survival of cell passes through MTT (3- (4,5- dimethyl -2- thiazolyls) -2,5- diphenyl -2H- tetramethyl azos azoles) Experiment is assessed.Cell is placed 24 hours on 96 orifice plates, with the extract-treated 48 of the crude drug composition of various concentration Hour.Then, cell is hatched 3 hours at 37 DEG C with MTT.Then, culture medium is abandoned, the additional 100 μ L into each hole DMSO.Plate is vibrated in the dark 15 minutes.Then, with ELIASA (Thermo Scientific companies, Freemont, Jia Lifu Buddhist nun Asia state) measure 570nm under absorbance.

(4) processing of the crude drug composition in extract

The effect of the extract for Neurite Outgrowth is investigated with the PC12 cells of culture.As display god Positive control through first projection provide 50ng/mL nerve growth factor (NGF) (from Alomone Labs, Ltd. buy, Cat#:N-100).To 6 orifice plates sowing PC12 cells (5x104 cells/wells).After 24 hours, with the HS containing 1% FBS, 1% And 1% P/S DMEM culture mediums have changed cell.In extraction process, a series of every 24 hours extracts to concentration (0.1,0.5 and 5mg/mL) is handled 2 times.Then, cell is collected for further experiment.

(5) measure of Neurite Outgrowth

After processing, using containing phase difference light collecting device and 10 times of object lens and digital camera (Diagnostic Instruments, Stirling highland, the state of Michigan) light microscope (Diagnostic Instruments) and pass through hand Dynamic setting captures image.In order to analyze the quantity of nervous process and length, to each culture medium from randomly selected at least ten The visual field counts about 100 cells.Then, using Photoshop (registration mark) softwares to cellular neural number of projection and length Degree is analyzed.When more than one nervous process is longer than the diameter of cell body, graduate division is carried out to cell, in addition, The length of nervous process according to possessed by cell body is categorized into different groups.That is, be divided into less than 30 μm, 30~60 μm, 60~ 90 μm and more than 90 μm several classes.In this experiment, NGF (50ng/mL) has been used as positive control.

(6) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)

After extraction process, by for the Protein Extraction of Neural Differentiation be less salt dissolve buffer solution (10mM HEPES, PH7.4,150mM NaCl, 1mM EDTA, 1mM EGTA, 0.5% Triton X-100,5mM benzamidine HCl, 10 μM Aprotinin, 10 μM of leupeptin).After being centrifuged 20 minutes at 4 DEG C with 13200rpm (16100 × g), supernatant is collected, Tot Prot is determined using protein test.All sample canonicals are turned to the albumen of same rank, with 2xSDS-PAGE samples Buffer solution (0.125M Tris-Cl, pH6.8,4% SDS, 20% glycerine, 2% 2 mercapto ethanol and 0.02% Bromophenol blue) handled, boiled 15 minutes before implementing SDS-PAGE.

(7) Western blot analysis

After electrophoresis, turn stain groove (Mini Trans-Blot (registration mark) CeLL) by Protein transfer using mini wet type To nitrocellulose membrane.Jump condition is as described below.1x transfering buffering liquids (25mM Tris, 192mM glycine, 15% second Alcohol, 0.1% SDS), 16 hours ,~40V and~0.1A.Dyed by using Ponceau S pair and the load of sample transfer equivalent Confirmed.At room temperature, in the Tween20 (TBS-T) of Tris buffer saline/0.1%, with 5% skimmed milk pair Nitrocellulose membrane blocks 2 hours, then, at 4 DEG C, in TBS-T, is once resisted using what is diluted in the skimmed milk to 2.5% Body hatches a Dinner.Then, nitrocellulose membrane is rinsed with TBS-T, at room temperature, in TBS-T, utilizes the skimmed milk to 2.5% The anti-mouse secondary antibodies of peroxidase (HRP) coupling of middle dilution or the anti-rabbit of peroxidase (HRP) coupling are secondary anti- Body is hatched 2 hours.After fully being cleaned with TBS-T, immunocomplex is visualized using enhanced chemiluminescence (ECL) method. According to the calibration curve that a serial dilution degree is built by Parallel gels in sample, compared using image dissector identical Gel on run under the conditions of the ECL for strictly regulating control and various samples in band intensity.

It is as follows to test the antibody used.

【Table 2】

(8) other are tested

Using the kit obtained from Bio-Rad companies (Heracles, California), according to Bradford Method determines protein concentration.Statistics experiment is carried out using a dimension deviation dispersion analysis.To being obtained from a reference value (Basal) or control value To difference classified as follows.

* p ＜ 0.05, * * p ＜ 0.01 and * * * p ＜ 0.001.

[result is with investigating]

(1)The effect of DTS extracts and YHK extracts to PC12 cells propagation

PC12 cells be such as NGF etc various stimulations can be reacted, can be abundant for Neural Differentiation The cell model of establishment.First, cell survival assay is carried out, determines that all extracts will not cause cell propagation dead with cell The safe concentration scope (0~5mg/mL) for each extract died.As a result, cell survival rate does not have under 0~5mg/mL concentration Have and significantly affected by DTS extracts and YHK extracts, be 0.1,0.5 and 5mg/mL of following experimental selection therefore (basic, normal, high) three kinds of various concentrations (Fig. 1).

(2)The biological evaluation of DTS extracts and YHK extracts to Neural Differentiation

PC12 cells are shown and spy as the sympathetic nervous metaclass of neurofilament expression and Neurite Outgrowth etc Sign.Herein, the quantity and the expression of length and neurofilament (NF68 and NF160) of the Neurite Outgrowths of PC12 cells are entered Investigation is gone, and morphology after the processing to DTS extracts and YHK extracts and biology change are investigated.

2-1)The Neurite Outgrowth of PC12 cells is induced during on DTS extracts and YHK extracts is used alone

Morphology investigation has been carried out to the Neural Differentiation of PC12 cells with length by the quantity for determining nervous process.This When, 50ng/mL NGF has been used as positive control.After being handled 48 hours with 50ng/mL NGF, it was observed that PC12 cells Significant Neurite Outgrowth.0.1,0.5 and 5mg/mL (concentration selected in previous experiment) DTS extracts or In the processing of YHK extracts, the obvious growth (Fig. 2A and B) of culture nervous process is not observed.However, as following experiment is tied Shown in fruit, YHK extracts induce the expression of the neurofilament of PC12 cells in higher concentrations, in addition, DTS extracts and YHK extractions Thing promotes the expression that the Neurite Outgrowth by NGF inductions induces with the NGF by neurofilament.

2-2)The expression that YHK extracts induce the neurofilament of PC12 cells in higher concentrations is used alone

Neural Differentiation can pass through the expression of the principal structural component neurofilament for the nerve cell (neuron) for parsing differentiation To carry out biochemical measurement.In the growth period of nervous process, mammiferous neurofilament subunit NF68 (about 68kDa) and NF160 When (about 160kDa) builds the domain of nervous process, heterodimer is formed.

If the 50ng/mL cultivated on PC12 cells NGF is applied into 48 hours, NF68 and NF160 two is reliably induced The expression of person.The DTS extracts on culture PC12 cells are used alone, under all concentration, not to F68 and NF160 table Up to being controlled.On the other hand, YHK extracts are used alone, under high concentration (5mg/mL), NF68 and NF160 can be raised Protein expression (about 4 times) (Fig. 3 A and B).

2-3)DTS extracts and YHK extracts promote the Neurite Outgrowth induced by NGF

In the present embodiment, express the NGF's of almost completely free effect as the induction to Neurite Outgrowth and neurofilament Appropriate concentration, select 0.5ng/mL NGF concentration.

In Neurite Outgrowth experiment, pass through DTS or YHK extracts (0.1,0.5 and 5mg/mL) and NGF (0.5ng/ ML the percentage for the cell that collaboration) is handled and broken up depends on usage amount.After DTS and NGF collaborations processing, show longer Nervous process, such as the PC12 cells of 30~60 μm, 60~90 μm and the nervous process more than 90 μm increased.To YHK With NGF cooperate with processing also carried out identical observation (Fig. 4 A and B).

2-4)DTS extracts and YHK extracts promote the expression of the neurofilament by NGF inductions

In order to further investigate the facilitation effect of DTS extracts and YHK extracts in Neural Differentiation, for PC12 cells pair DTS extracts or YHK extracts (0.1,0.5 and 5mg/mL) have carried out cooperateing with processing with NGF (0.5ng/mL).DTS is all Collaboration processing is carried out under concentration together with NGF, as a result, NF68 expression is slightly increased, in addition, 0.1 and 0.5mg/mL's YHK is cooperateed with processing with NGF's, and NF68 is expressed as 2~3 times.5mg/mL YHK and NGF cooperate with processing in, it was observed that NF68 obvious increase (about 8 times).NF160 expression becomes apparent from all groups than NF68 increases.In cooperateing with for DTS and NGF In processing, 0.1mg/mL DTS shows highest effect (about 6 times) to NF160 expression.Processing is cooperateed with YHK and NGF In, 5mg/mL YHK shows highest effect (about 9 times) to NF160 expression.Reference picture 5A and B.

[conclusion]

In the case where DTS extracts and YHK extracts is used alone, if be used together with NGF, even if with NGF certainly The NGF of the body not low usage amount of display effect, it can also promote the growth of inducing neural projection, the expression of increase neurofilament.

Embodiment 2

The subjects for making to have taken the DTS or YHK of embodiment 1 within certain period answer application form, verify and dull-witted The related effect of the invention of disease prevention and treatment.

As the application form for simply being checked dementia, as described above, employing Fig. 6 big friendly formula dementia Prediction test.

Subjects as respondent are that to take more than 3 years persons of both YHK or YHK and DTS (most long to take experience For 15 years).But dysfunction of liver all once occurred for all subjects (once with hepatitis C person 80%, other liver functions Abnormal person 20%), it is unknown to suffer from other chronic ill persons.

Method：Application form is distributed to subjects, subjects are returned after filling in.

Period：Optionally filled in before 2 the end of month in 2017, give March and reply.

Subjects are total 62, and the subjects number of every individual character Do and age range section is as shown in Figure 7.

Make following judgement to score substantially, 0~8 point for it is normal, 9~13 points to should be noted, 14~20 points is can Dementia early symptom can occur.

Result is summarized as bar chart, as shown in Fig. 8~Figure 13.

1. the analysis related to taking year

(1) in YHK is only taken, only take DTS, take YHK+DTS complete group of subjects, do not show The subjects of dementia early symptom.

(2) in only YHK group is taken, as shown in figure 8, take had 50% for it is noted that taking 3 years less than 3 years The above, take the whole of more than 6 years normally.

(3) in only DTS group is taken, as shown in figure 9, the user for taking 3 years to 6 years has 50% for it is noted that 6 User more than year is all normal.

(4) it is as shown in Figure 10, also all normal even if less than 3 years in both YHK and DTS is taken.Although there are clothes There are 18% (in 11 2) it is noted that but this time age distribution of investigation is close to higher limit 76~80 years old with 3~6 years Age group, it can be considered to receive age increased influence.

2. the analysis related to the age of subjects

(1) YHK subjects are only taken, as shown in figure 11, as long as being normal known to age bracket.

(2) in only DTS subjects are taken, as shown in figure 12, occur should be noted in the group of 61~65 years old Patient's (all 33%).

(3) it is as shown in figure 13, overall in the group of 76~80 years old in the subjects for taking both YHK and DTS 6% it is noted that other are all normal.

According to the above results, in only more than YHK3 subjects are taken, there are not dementia early stage and needs The patient paid attention to.It may determine that, if only taking DTS, more than the 6 years long-term use of knots that can then obtain improvement hepatic and renal function Fruit, easily activate brain cell.In YHK and DTS person is used in combination, also there is not dementia less than 3 years even if year is taken Early stage and the patient that should be noted are all normal.It is possible thereby to judge, YHK or YHK's and DTS is used in combination to dementia Prevention be effective.

As described in the paragraph in background technology, the People of more than 70 years old is set to be checked for renewal driving license, Police agency announce data (http://www.zensiren.or.jp/kourei/data/data.html) in, the 1st class (memory and judgment reduction person) is that the 2.3%, the 2nd class (memory and judgment slightly reduction person) is the 26.7%, the 3rd class (memory with judgment completely do not have no problem person) is 71.0%.Compared with the data, in the present embodiment, taking Both YHK, DTS or YHK and the DTS subjects more than 70 years old add up in 26 people, it is necessary to which the person of attention is (equivalent to the above-mentioned 2nd Class) it is that 2 people account for 7.7%, normal (equivalent to above-mentioned 3rd class) accounts for 92.3% for 24 people, it follows that the dementia of the present invention is pre- It is anti-good with therapeutic effect.

Claims

A kind of 1. crude drug composition, for preventing either to treat dementia or its behavior and mental symptoms or nerve god Through degenerative disease, it is characterised in that contain pseudo-ginseng and the bark of eucommia as crude drug composition.
2. crude drug composition according to claim 1, it is characterised in that also contain sealwort as crude drug composition.
3. crude drug composition according to claim 2, it is characterised in that also contain radix glycyrrhizae as crude drug composition.
4. crude drug composition according to claim 1, it is characterised in that the pseudo-ginseng is 10~90 weight %, Du Secondary is 10~90 weight %.
5. crude drug composition according to claim 2, it is characterised in that the pseudo-ginseng is 10~90 weight %, Du Secondary is 10~90 weight %, and the sealwort is 2~25 weight %, limits each crude drug component content sum as 100 weight %.
6. crude drug composition according to claim 3, it is characterised in that the pseudo-ginseng is 10~90 weight %, Du Secondary is 10~90 weight %, and the sealwort is 2~25 weight %, and the radix glycyrrhizae is 3~20 weight %, limits each crude drug composition and contains Amount sum is 100 weight %.
7. crude drug composition according to claim 3, it is characterised in that the pseudo-ginseng is 10~90 weight %, Du Secondary is 10~90 weight %, and the sealwort is 2~25 weight %, and the radix glycyrrhizae is 5~20 weight %, limits each crude drug composition and contains Amount sum is 100 weight %.
8. the crude drug composition according to any one of claim 1 to claim 7, it is characterised in that the crude drug Composition is the extract of each crude drug composition.
9. the crude drug composition according to any one of claim 1 to claim 7, it is characterised in that the dementia Disease or its behavior and the prevention of mental symptoms or nerve degenerative diseases or treatment are by increasing the expression of neurofilament or promoting god Carried out through the growth of projection.
10. crude drug composition according to claim 8, it is characterised in that the dementia or its behavior and mental symptoms Or the prevention or treatment of nerve degenerative diseases are carried out by the expression for increasing neurofilament or the growth for promoting nervous process.