CN104800303B - It is a kind of to be used to treat Chinese medicine composition of Alzheimer disease and its preparation method and application - Google Patents
It is a kind of to be used to treat Chinese medicine composition of Alzheimer disease and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of Chinese medicine composition for being used to treat Alzheimer disease and its preparation method and application.The Chinese medicine composition of the present invention is by the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe and fructus polygoni orientalis composition, research shows that the Chinese medicine composition of the present invention can improve the Spatial learning ability and space exploration ability of Alzheimer disease kunming mice, improve the brain of Alzheimer disease mouse and the organ index of spleen, reduce the permeability of Alzheimer disease blood-brain barrier, Alzheimer disease brain tissue hippocampus A β, IL 6, the protein expression levels of IL 12 are reduced, and on Alzheimer disease model kunming mice body weight without obvious influence.It these results suggest that the Chinese medicine composition of the present invention is good for the therapeutic effect of Alzheimer disease, therefore the Chinese medicine composition of the present invention will have broad application prospects in treatment of alzheimer field.
Description
Technical field
It is more particularly to a kind of to be used to treat A Erci the present invention relates to a kind of Chinese medicine composition and its production and use
Chinese medicine composition of the silent disease in sea and its preparation method and application, the invention belongs to traditional Chinese medicine research technical field.
Background technology
Alzheimer disease is a kind of one of common geriatric disease, and disturbance of intelligence occurs in the brain tissue for showing as body
And its phrenoblabia, this is a kind of comprehensive, multiple nervous system degenerative disease.The pathology table of Alzheimer disease
Now occurs the deposition of substantial amounts of amyloid plaques for brain tissue, before the main component of patch is amyloid-beta (A β), A β
Body is amyloid-beta precursor protein (APP), and A β are that APP hydrolyzes a kind of 40-42 ammonia produced by α, β, gamma-secretase
The peptide matters of base acid, mainly include A β1-40With A β1-42, wherein A β1-40Toxicity there is no A β1-42Greatly, based on this, experiment can be with
Using A β1-42Or A β1-40Alzheimer disease model is set up in brain tissue injection, carries out the experimental study to Alzheimer disease.
Alzheimer disease model brain tissue hippocampus A β horizontal abnormalities, Alzheimer disease model brain tissue can be observed based on this simultaneously
Hippocampus A β expression.Occur exception more Alzheimer disease model behavioral indexes, be so accomplished by detecting alzheimer '
The Spatial learning ability and its space exploration ability of silent disease model, judge the learning and memory ability of Alzheimer disease model,
Based on this, it is necessary to detect the behavioral indexes of Alzheimer disease model.
Zlokovic proposes Alzheimer disease neural blood vessel hypothesis, it is believed that blood-brain barrier, which changes, causes a variety of cascades anti-
Should, the permeability changes influence brain tissue hippocampus of blood-brain barrier removes the (Zlokovic such as abnormal cause A β depositions to A β
BV.Neurovascular mechanisms of Alzheimer’s neurodegeneration[J].Trends
Neurosci, 2008,28 (4)：202-208.), and the permeability of blood-brain barrier can by detect brain tissue EB content come
Display (Wang Ping, Lin Yang, Ge Feifei, wait influence [J] dissection scientific advancees of the .CRM197 to cavy blood-brain barrier permeability,
2008,14 (4)：416-419.), based on this, the brain tissue EB contents of Alzheimer disease model can be detected.Alzheimer
The decline of sick many performance immune responses, can cause the expression of substantial amounts of immune factor increases, the factor such as including IL-6, IL-12
Increase, based on this can by determine IL-6, IL-12 protein expression level judge Alzheimer disease immune depression react
(Richartz E,Stransky E,Batra A, et al.Decline of immune responsiveness：a
pathogenetic factor In Alzheimer's disease[J].J Psychiatr Res.2005Sep；39(5)：
535-543.Swardfager W, Rothenburg L, et al.A meta-analysis of
cytokines in Alzheimer's disease[J].Biol Psychiatry.2010Nov 15；68(10)：930-
941.Lin nW,Levison SW.Context-dependent IL-6potentiation of interferon-gamma-
induced IL-12secretion and CD40expression in murine microglia[J].J Neurochem.2009Nov；111(3)：808-818.).
The Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe is one of composition of the ancient prescription Buyang Huanwu Tang of motherland.Buyang Huanwu Tang is motherland's medical science
One of ancient prescription, the present inventor's previous work points out that Buyang Huanwu Tang and the side of tearing open are big to Alzheimer disease caused by A β 1-40
Mouse serum can play adjustment effect with hippocampal tissue A β and cytokine content, and (Zhou Zhongguang, Han Yusheng, Jiang Bo wait yang-tonifyings also
Five soup and the side of tearing open are to A β1-40Caused Alzheimer's Disease Rats serum and hippocampal tissue A β and influence [J] of cytokine content
Shanxi Medicine magazine, 2008,37 (2)：129-131.), on this basis, inventor is changed to Buyang Huanwu Tang
Good, i.e., a kind of Chinese medicine composition for being used to treat Alzheimer disease of the invention is except the fractions containing Buyang Huanwu Tang
Outside the Radix Astragali, Radix Angelicae Sinensis and the radix paeoniae rubrathe, the also medical fructus polygoni orientalis of motherland, wherein fructus polygoni orientalis has antitumor, immunosupress, antioxygen
Change, anti-hepatic fibrosis etc. are acted on.
It is demonstrated experimentally that compared to ancient prescription Buyang Huanwu Tang, Radix Astragali dosage is reduced about in Chinese medicine composition of the invention
2/3rds, it but can still reach the therapeutic effect same or like with Buyang Huanwu Tang.Current Chinese medicine basic research is than relatively thin
Weak, chemical composition complex in Chinese medicine material medicine, effective component of chinese medicine is indefinite, and Chinese medicine is multicomponent Mutiple Targets synthesis hair
The effect of waving, therefore, flavour of a drug are more more more easily to cause adverse reaction.In consideration of it, the present inventor has simplified Chinese prescription, only select yellow
Stilbene, Radix Angelicae Sinensis, the radix paeoniae rubrathe and fructus polygoni orientalis carry out prescription, and prepare alzheimer ' using A β 1-42 injection kunming mices hippocampus
Silent disease animal model, has carried out the research to preventing and treating Alzheimer disease.
The content of the invention
The technical problems to be solved by the invention be to provide it is a kind of be used for treat Alzheimer disease Chinese medicine composition and
Its preparation method and application.
A kind of Chinese medicine composition for being used to treat Alzheimer disease of the present invention, by each bulk drug group of following parts by weight
Into：Radix Astragali 30-50 parts by weight, Radix Angelicae Sinensis 4-8 parts by weight, radix paeoniae rubrathe 3-7 parts by weight and fructus polygoni orientalis 15-25 parts by weight.
In the present invention, it is preferred to, described Chinese medicine composition is made up of each bulk drug of following parts by weight：The weight of the Radix Astragali 40
Measure part, the parts by weight of Radix Angelicae Sinensis 6, the parts by weight of the radix paeoniae rubrathe 5 and the parts by weight of fructus polygoni orientalis 20.
Further, the invention also provides a kind of clinically suitable Chinese medicine system for being used to treat Alzheimer disease
Agent, it is characterised in that as the auxiliary material needed for described Chinese medicine composition adds preparations shaping, according to the routine for preparing pharmaceutical preparation
Method is made.
In the present invention, it is preferred to, described preparation is water decoction, capsule, pill, granule, tablet or oral liquid.
The preparation method of invention formulation belongs to conventional formulation method, and there is no need to go into details herein.
Further, the invention also provides a kind of method for preparing described Chinese medicine composition, it is characterised in that bag
Include following steps：
(1) parts by weight as described above weigh each bulk drug；
(2) mixture of each bulk drug is put into high neck round-bottom flask, adds the distilled water of 7-10 times of medicinal material gross weight,
Glass bar is stirred, and 1.5-2.5h is soaked under enclosed environment, and heating is decocted after 1.5-2.5h, cooling 10min.Use multilayer gauze
Medicinal material composition is filtered, until medicine does not have decoction outflow, first time filtrate is obtained；
(3) medicine residue is put into high neck round-bottom flask again, adds crude drug 7-10 times of distilled water of gross weight, repeat to walk
Suddenly the process of (2) carries out second of filtration treatment, obtains second of filtrate, merges filtrate twice；
(4) filtrate obtains the extraction concentrate of Chinese medicine composition after 70 DEG C of evaporation moisture of bitubular steamer, and what is obtained carries
Concentrate is taken to continue to dry 4-5d in 70 DEG C of electric heating constant-temperature blowing drying boxes, resulting residue is exactly described Chinese medicine group
The dry cream of compound.
Further, also include the dry cream of Chinese medicine composition being placed in pulverizer in the process described above crushing, enter
Row grinding, molecular sieve processing, obtains the pulvis of the Chinese medicine composition repeatedly.
It is preferred that, in described method, the parts by weight of each bulk drug are：The parts by weight of the Radix Astragali 40, the parts by weight of Radix Angelicae Sinensis 6,
The parts by weight of the radix paeoniae rubrathe 5 and the parts by weight of fructus polygoni orientalis 20.
The present invention has carried out using kunming mice as process object, and the micro sample adding appliance bilateral ventricles of the brain are disposably noted under aseptic condition
Enter 5 μ l condensed state A β1-42(80pmol μ l-1), prepares Alzheimer disease model.Using by Radix Astragali 40g, Radix Angelicae Sinensis 6g, the radix paeoniae rubrathe
The Chinese medicine composition of 5g, fructus polygoni orientalis 20g composition, the high, medium and low dosage 18.46gkg of setting Chinese medicine composition-1·d-1、
9.23g·kg-1·d-1、4.62g·kg-1·d-1Three dosage groups carry out the experimental study to Alzheimer disease, are easy to observation
The content of influence of the Chinese medicine composition to Alzheimer disease model kunming mice, main detection and test includes Alzheimer
The Spatial learning ability and space exploration ability of sick kunming mice, brain organ index, the organ index of spleen, blood-brain barrier it is logical
The body weight of permeability, brain tissue hippocampus A β, IL-6, IL-12 protein level and its Alzheimer disease model kunming mice, as a result table
Bright, Chinese medicine composition of the invention can improve the learning and memory function of Alzheimer disease model mouse, and reduce inflammation reaction, drop
The permeability of low blood-brain barrier, the Chinese medicine composition can be by suppressing hippocampus A β and other inflammatory factor albumen in A Erci
Played an important role in the silent disease treatment in sea, while aβ protein has correlation with the change of IL-6, IL-12 protein content.More than
The comprehensive Chinese medicine composition for showing the present invention is good to the combined therapy effect of Alzheimer disease.
Therefore, on this basis, the invention also provides described Chinese medicine composition is preparing treatment Alzheimer disease
Purposes in medicine.
Brief description of the drawings
Influences of the Fig. 1 for Chinese medicine composition of the invention to Alzheimer disease mouse orientation navigation ability；
Influences of the Fig. 2 for Chinese medicine composition of the invention to Alzheimer disease mouse space exploration ability；
Influences of the Fig. 3 for Chinese medicine composition of the invention to Alzheimer disease mouse brain organ index；
Influences of the Fig. 4 for Chinese medicine composition of the invention to Alzheimer disease mouse spleen organ index；
Influences of the Fig. 5 for Chinese medicine composition of the invention to Alzheimer disease Mice brain tissues EB contents；
The influence that Fig. 6 is expressed Alzheimer disease mouse brain hippocampal tissue aβ protein for the Chinese medicine composition of the present invention；
Fig. 7 is the shadow of Chinese medicine composition of the invention to Alzheimer disease mouse brain hippocampal tissue IL-6 protein expressions
Ring；
Fig. 8 is the shadow of Chinese medicine composition of the invention to Alzheimer disease mouse brain hippocampal tissue IL-12 protein expressions
Ring；
Influences of the Fig. 9 for Chinese medicine composition of the invention to Alzheimer disease mouse weight.
Embodiment
Below by experiment and the present invention will be further described in conjunction with the embodiments, it should be understood that these embodiments
The purpose of illustration is only used for, protection scope of the present invention is never limited in.
A kind of preparation for being used to treat the Chinese medicine composition of Alzheimer disease of embodiment 1
The Chinese medicine composition of the present invention is made up of each bulk drug of following weight：Radix Astragali 40g, Radix Angelicae Sinensis 6g, radix paeoniae rubrathe 5g and
Fructus polygoni orientalis 20g.
A kind of preparation for being used to treat the Chinese medicine composition of Alzheimer disease of embodiment 2
The Chinese medicine composition of the present invention is made up of each bulk drug of following weight：Radix Astragali 35g, Radix Angelicae Sinensis 8g, radix paeoniae rubrathe 4g and
Fructus polygoni orientalis 24g.
A kind of preparation for being used to treat the Chinese medicine composition of Alzheimer disease of embodiment 3
The Chinese medicine composition of the present invention is made up of each bulk drug of following weight：Radix Astragali 45g, Radix Angelicae Sinensis 5g, radix paeoniae rubrathe 7g and
Fructus polygoni orientalis 14g.
A kind of preparation for being used to treat the Chinese medicine composition (dry cream) of Alzheimer disease of embodiment 4
Testing the medicinal material used includes Radix Astragali 40g, Radix Angelicae Sinensis 6g, radix paeoniae rubrathe 5g, fructus polygoni orientalis 20g, and medicinal material is uniformly mixed to get
71g, prepares 10 this secondary medicinal material according to same compound method, obtains 710g medicinal material total amount.The mixture of 710g medicinal materials is put
In the high neck round-bottom flask for entering 20000ml, 6000ml distilled water is added, glass bar is stirred, and 2h is soaked under enclosed environment, plus
Heat decocts 2h, 10min is cooled down, with multilayer filtered through gauze medicinal material composition, filtered through gauze residue, until medicine does not have decoction stream
Go out, obtain first time filtrate.Medicine residue is put into high neck round-bottom flask again, 6000ml distilled water is added, mistake above is repeated
Second of filtration treatment of Cheng Jinhang, obtains second of filtrate, merges filtrate twice.
Filtrate obtains the extraction concentrate of Chinese medicine composition after 70 DEG C of evaporation moisture of bitubular steamer, and obtained extraction is dense
Contracting liquid continues to dry 4-5d in 70 DEG C of electric heating constant-temperature blowing drying boxes, and resulting residue is exactly described Chinese medicine composition
Dry cream.
Experiment obtains Chinese medicine composition dry cream 267.1g, and Chinese medicine composition paste-forming rate is dry cream/Chinese medicinal material × 100%=
37.62%.
A kind of preparation for being used to treat the Chinese medicine composition (pulvis) of Alzheimer disease of embodiment 5
The Chinese medicine composition dry cream that Example 4 is obtained, Chinese medicine composition dry cream is placed in pulverizer and crushed, is ground
Mill, repeatedly molecular sieve processing reduces and obtains Chinese medicine composition dry powder 261.5g, Chinese medicine composition after being crushed in loss, pulverizer
Flour extraction be dry powder/Chinese medicinal material × 100%=36.83%.The medicinal material of actual use is 710g, and obtained dry powder is
261.5g, therefore 1g Chinese medicine compositions dry powder is equivalent to active compound medicinal material 2.72g.
Chinese medicine composition dry powder 10g packing is weighed after electronic balance zeroing, sealing is placed in -80 DEG C of refrigerators to be protected for a long time
Deposit.
Purposes of the Chinese medicine composition of the present invention of embodiment 6 in treatment Alzheimer disease
1. material and instrument
1.1 material
54 males, 8 week old, cleaning grade kunming mice, 25 ± 3g of body weight, Heilongjiang University of Chinese Medicine's Experimental Animal Center
Li Baolong is provided, certification of fitness：SCXK (black) 2013-004.Kunming mice is in Heilongjiang University of Chinese Medicine experimental animal
Heart normal temperature, it is often wet, sterile under appropriate conditions of raise, kunming mice freely absorbs food and drinking water.Piracetam buy in
Hunan Dino Pharmaceutical Co, drug batch number：130325.A β 1-42 are purchased from Sigma companies.A β, IL-6, IL-12 are enzyme-linked to exempt from
Epidemic disease kit is bought builds up Bioengineering Research Institute, lot number in Nanjing：20140401.Distilled water is bought to be steamed in the literary scape in Harbin City
Distilled water factory.Radix Astragali lot number 131109, Radix Angelicae Sinensis lot number 131001, radix paeoniae rubrathe lot number 121003, fructus polygoni orientalis lot number 140922, by black dragon
Chinese medicine study institute of river university of TCM professor Zhou Zhongguang is bought in the attached First Hospital of Heilongjiang University of Chinese Medicine.Chinese traditional medicine composition
Thing dry powder is prepared according to the method for embodiment 5, is given when mouse stomach is treated, it is necessary to by medicinal material dry composition into equivalent to original
Medicinal material 18.46gkg-1·d-1、9.23g·kg-1·d-1、4.62g·kg-1·d-1Three dosage, and to Chinese medicine composition
Progress is heated to 37 DEG C, is easy to give mouse stomach.Other reagents are that domestic analysis is pure.
1.2 instrument
HMIAS high-definition color medical science picture and text analysis systems, the production of the screen image technology Co., Ltd of Wuhan thousand；RE-52A
Rotary Evaporators, Shanghai Yarong Biochemical Instrument Plant's production；SIMF124 ice machines, the production of SANYO GS company；CM1900 frosts are cut
Piece machine, the production of Leica companies；PLZOZ-S electronic balances, plum Teller-support benefit Instrument Ltd. production；Morris water fans
Palace, the production of Northern Huaihe River Anhui Zheng Hua biological Instrument and equipments Co., Ltd；6100 type RT- thunders Du's ELIASA, the production of RT companies of the U.S.；
TGL-16G desk centrifuges, Anting Scientific Instrument Factory, Shanghai's production；Superclean bench, the limited public affairs of Suzhou Bo Laier cleaning equipments
Department's production；Alcohol meter, the production of glass factory of Hejian City of Hebei province；101-0AB type electric heating constant-temperature blowing drying boxes, Tianjin Stettlen
Instrument Ltd.'s production production；Autoclave, Shanghai Medical Nuclear Instrument Factory's production etc..
2 methods
2.1 experiment packet
Kunming mice divides control group 9, model group 9, Piracetam group 9, Chinese medicine composition high dose group at random
(18.46g·kg-1·d-1) 9, Chinese medicine composition middle dose group (9.23gkg-1·d-1) 9, Chinese medicine composition low dosage
Group (4.62gkg-1·d-1) 9, mouse totally 54.
2.2 Alzheimer disease models are replicated
The general state of kunming mice is observed, kunming mice, preference pattern group, Piracetam group, Chinese medicine composition is captured
High dose group, Chinese medicine composition middle dose group and Chinese medicine composition low dose group mouse are small to Kunming using 10% chloraldurate
Mouse is anaesthetized, fixed mouse, and micro sample adding appliance disposably injects 5 μ l condensed states A in the bilateral ventricles of the brain of mouse under aseptic condition
β1-42(80pmol μ l-1), syringe needle leaves 2min, wraps up mouse, allows mouse fully to rest, and prevents infection.Control group is adopted simultaneously
With same method the bilateral intracerebroventricular equivalent of mouse physiological saline.
2.3 experimental administration
Radix Astragali 40g, Radix Angelicae Sinensis 6g, radix paeoniae rubrathe 5g, fructus polygoni orientalis 20g dosage determine that foundation is：Normal body standard weight
Drug dose is 71g, and by the ratio between body surface area of people and mouse, the drug dose for trying to achieve every 20g mouse standard weight is 71g
0.0026 is multiplied by, the drug dose 0.1846g/20g of every 20g mouse standard weight is obtained, is converted into 9.23g/kg, this dosage is just
It is the middle dosage for the mouse medication tested, heavy dose is doubled compared with middle dosage, low dose halves compared with middle dosage.
Control group, model group mouse give the physiological saline gavage of equivalent, and Piracetam group gives Piracetam 0.62g
kg-1·d-1Gavage, Chinese medicine composition high dose group gives Chinese medicine composition 18.46gkg-1·d-1In gavage, Chinese medicine composition
Dosage group gives Chinese medicine composition 9.23gkg-1·d-1Gavage, Chinese medicine composition low dose group give Chinese medicine composition
4.62g·kg-1·d-1Gavage, the gavage duration is 21d.
2.4 test position fix operational capabilities and space exploration ability
The capital equipment of Morris water mazes test includes：Morris water mazes, computer camera system and software analysis system
System.Each group mouse orientation navigation ability and space exploration ability, i.e. each group mouse is detected using Morris water mazes instrument
Habit ability and memory capability.The Morris water mazes instrument is circular pond, inside contains a transportable platform, platform
On have band, mouse can be facilitated to stop, the shooting instrument above pond is connected with computer, at any time to the movement locus of mouse
It is tracked, analyzes the corresponding experimental data of mouse.Need to keep the object of reference of indoor and outdoor consistent when test, reduce extraneous
Interference.Morris water maze instrument depth of water 20cm, temperature control puts prepared Chinese ink in (23 ± 2) DEG C, in water and forms muddy black
Background, because mouse is white, it is necessary to manufacture black background, is easy to carry out and tests.
Four quadrants are determined by imaging instrument.Wherein Morris water mazes platform is put in the fourth quadrant of water maze, separately
Outer 3 quadrants as test quadrant.Test the 1st, 2,3,4d be the training time, 60s is trained for every time, will be small when test
First, second and third quadrant that mouse head is put in Morris water maze instruments towards outside respectively carries out experimental study, records each mouse
Swimming track and its corresponding experimental data.Test 5d test mouse and reach the underwater platform position time when being denoted as incubation period
Between, the orientation navigation ability of mouse is weighed with this.Morris water maze instrument platforms are removed when 5d is tested in addition,
Detection mouse in 60s finds the swimming distance of platform as mouse space exploration ability is weighed, and the note of mouse is weighed with this
Recall ability.
2.5 determine organ index
Morris water maze test experiments terminate after 2h, and ice platform is placed on superclean bench, is operated, rapid broken end
Dead each group mouse, brain, the spleen organ of each group mouse of drawing materials, electronic balance accurately weighs the Organ weight of mouse brain, spleen, entered
Row statistical analysis, calculates mouse brain, the organ index of spleen.
The calculation formula of mice organs index：Organ index (mgg-1)=Organ weight (mg)/body weight (g)
2.6 determine brain tissue Azo-Blue (EB) content
Experiment is made into EB standard liquids according to coubling dilution successively, and concentration is respectively 8,4,2,1,0.5,0.25 μ g
ml-1, 630nm absorbance (A) is detected on 6100 type RT- thunders Du's ELIASA.Using EB concentration as abscissa, absorbance
(A) it is ordinate, draws the standard curve of EB contents, seek EB content standard curvilinear equations.
Morris water maze test experiments terminate after 2h, the tail vein injection 2%EB solution of each group mouse, observe at any time small
The whole skin of mouse occurs after obvious blueness, shows that 2%EB solution is injected successfully.Mouse water after 2%EB solution is injected successfully
Close chloral to be anaesthetized, cord fixes mouse, 100ml physiological saline irrigated to the left ventricle of mouse using sterile transfusion device,
Dissection, formamide solution immersion, lucifuge are incubated, are homogenized, centrifuge 10min rapidly on ice platform.Use 6100 types of RT companies of the U.S.
Supernatant absorbance (A) is determined at RT- thunders Du's ELIASA, selection 630nm.EB concentration (μ g are obtained according to EB standard curves
ml-1), obtain EB contents (the μ gg of mouse in the quality by Mice brain tissues-1), EB contents EB concentration/brain wet weight (μ
g·g-1) ratio represent.
2.7 determine Hippocampus of Mice A β, IL-6 and IL-12 protein contents
Morris water maze test experiments terminate after 2h, each group mouse sacrificed by decapitation on superclean bench, remove brain group
Skin is knitted, cranium is opened, complete mouse brain tissue is taken out and is placed on ice platform, remove brain stem and cerebellum, each group mouse is taken rapidly
Hippocampal tissue, is homogenized Hippocampus of Mice, and 4 DEG C of refrigerators are stood, 5000rpmmin-110min is centrifuged, hippocampal tissue supernatant is drawn
Liquid, packing, -20 DEG C of preservations.Operated by enzyme linked immunological kit specification, use 6100 type RT- thunders Du's ELIASA 450nm
Each hole absorbance (A) is determined at wavelength, the content of mouse brain hippocampal tissue A β, IL-6 and IL-12 albumen is tried to achieve.
2.8 determine body weight
Mouse weight is determined during experiment daily, experiment finally determines kunming mice body weight before terminating, small with what is finally surveyed
Mouse body weight is Exemplary statistical data, calculates the weight differences for comparing each group mouse.
2.9 statistical analysis
Statistical procedures are carried out using SPSS19.0 softwares, experimental data is usedRepresent, multigroup is compared using single
Analysis of variance experimental data.P<0.05 thinks that difference is statistically significant.
3 results
Influence of 3.1 Chinese medicine compositions to Alzheimer disease mouse orientation navigation ability
Mouse orientation navigation ability is represented with incubation period.As a result show, compared with control group, model group mouse incubation period is bright
Aobvious extension (P ＜ 0.05)；Compared with model group, Piracetam group, Chinese medicine composition high, medium and low dosage group mouse incubation period are bright
It is aobvious to shorten (P ＜ 0.05) (being shown in Table 1, Fig. 1).
Influence of the Chinese medicine composition of table 1 to Alzheimer disease mouse orientation navigation ability
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.2 Chinese medicine compositions to Alzheimer disease mouse space exploration ability
Mouse space exploration ability is represented with swimming distance.As a result show, compared with control group, model group mouse swimming away from
From obvious increase (P ＜ 0.05)；Compared with model group, the high, medium and low dosage group mouse swimming of Piracetam group, Chinese medicine composition
Distance is significantly reduced (P ＜ 0.05) (being shown in Table 2, Fig. 2).
Influence of the Chinese medicine composition of table 2 to Alzheimer disease mouse space exploration ability
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.3 Chinese medicine compositions to Alzheimer disease mouse brain organ index
Analyzed, compared with control group from table 3, model group Alzheimer disease mouse brain organ index substantially reduces (P
＜ 0.05), compared with model group, the high, medium and low dosage group Alzheimer disease mouse brain of Piracetam group, Chinese medicine composition is dirty
Device index is significantly raised (P ＜ 0.05) (being shown in Table 3, Fig. 3).
Influence of the Chinese medicine composition of table 3 to Alzheimer disease mouse brain organ index
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.4 Chinese medicine compositions to Alzheimer disease mouse spleen organ index
Analyzed, compared with control group from table 4, the organ index of model group Alzheimer disease mouse spleen substantially drops
Low (P ＜ 0.05), is compared with model group, and the internal organs of the high, medium and low dosage group mouse spleen of Piracetam group, Chinese medicine composition refer to
Number significantly raised (P ＜ 0.05) (being shown in Table 4, Fig. 4).
Influence of the Chinese medicine composition of table 4 to Alzheimer disease mouse spleen organ index
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.5 Chinese medicine compositions to Alzheimer disease Mice brain tissues EB contents
Analyzed, compared with control group from table 5, model group Alzheimer disease Mice brain tissues EB contents are significantly raised
(P ＜ 0.05)；Compared with model group, the high, medium and low dosage group Alzheimer disease mouse brain of Piracetam group, Chinese medicine composition
Tissue EB contents are substantially reduced (P ＜ 0.05) (being shown in Table 5, Fig. 5).
Influence of the Chinese medicine composition of table 5 to Alzheimer disease Mice brain tissues EB contents
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
The influence that 3.6 Chinese medicine compositions are expressed Alzheimer disease mouse brain hippocampal tissue aβ protein
Analyzed, compared with control group from table 6, model group mouse Alzheimer disease cerebral hippocampal tissue aβ protein content
Significantly raised (P ＜ 0.05)；Compared with model group, the high, medium and low dosage group cerebral hippocampal tissue A of Piracetam group, Chinese medicine composition
β protein contents are substantially reduced (P ＜ 0.05) (being shown in Table 6, Fig. 6).
The influence that the Chinese medicine composition of table 6 is expressed Alzheimer disease mouse brain hippocampal tissue aβ protein
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.7 Chinese medicine compositions to Alzheimer disease mouse brain hippocampal tissue IL-6 protein expressions
From table 7, compared with control group, model group Alzheimer disease Hippocampus of Mice IL-6 protein contents are obvious
Raise (P ＜ 0.05)；Compared with model group, the high, medium and low dosage group Alzheimer disease of Piracetam group, Chinese medicine composition is small
Mouse hippocampal tissue IL-6 protein contents are substantially reduced (P ＜ 0.05) (being shown in Table 7, Fig. 7).
Influence of the Chinese medicine composition of table 7 to Alzheimer disease mouse brain hippocampal tissue IL-6 protein expressions
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.8 Chinese medicine compositions to Alzheimer disease mouse brain hippocampal tissue IL-12 protein expressions
From table 8, compared with control group, model group Alzheimer disease Hippocampus of Mice IL-12 protein contents are bright
Aobvious rise (P ＜ 0.05)；Compared with model group, the high, medium and low dosage group Alzheimer disease of Piracetam group, Chinese medicine composition
Hippocampus of Mice IL-12 protein contents are substantially reduced (P ＜ 0.05) (being shown in Table 8, Fig. 8).
Influence of the Chinese medicine composition of table 8 to Alzheimer disease mouse brain hippocampal tissue IL-12 protein expressions
Note：Compared with control group：1)P ＜ 0.05；Compared with model group：2)P ＜ 0.05
Influence of 3.9 Chinese medicine compositions to mouse weight
As a result show, compared with control group, the body weight of model group mouse changes not substantially (P ＞ 0.05)；With model group ratio
Compared with the mouse weight of the high, medium and low dosage group of Piracetam group, Chinese medicine composition changes not substantially (P ＞ 0.05) and (is shown in Table 9, figure
9)。
Influence of the Chinese medicine composition of table 9 to Alzheimer disease mouse weight
Note：Compared with control group：1)P ＞ 0.05；Compared with model group：2)P ＞ 0.05
4 conclusions
The ethological testing result of this zoopery shows that the ability of learning and memory of model group mouse is decreased obviously, mould
Type is stable, suitably carries out experimental study.The ability of learning and memory of the high, medium and low dosage group mouse of Chinese medicine composition is obviously improved, and is carried
Show that the high, medium and low dosage of Chinese medicine composition can be obviously improved the ability of learning and memory of Alzheimer disease model mouse.Say above
Bright Radix Astragali 40g, Radix Angelicae Sinensis 6g, radix paeoniae rubrathe 5g, fructus polygoni orientalis 20g Chinese medicine compositions can improve Alzheimer disease kunming mice space
Learning ability and space exploration ability.
Measuring mouse spleen, the organ index of brain show that the mass fraction of model group mouse brain and spleen is abnormal,
Mouse brain and splenic injury are heavier, and body brain function weakens, and anti-infection ability weakens.The high, medium and low dosage group of Chinese medicine composition can
To improve the brain of mouse and the mass fraction of spleen, strengthen the anti-infection ability of body, point out the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe, prince ' s-feather
Sub- Chinese medicine composition recovers spleen, the partial function of brain, has to the effect of Alzheimer disease model mouse corrosion disease and its brain function
Certain restitution.The medical Radix Astragali of motherland described above, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis crude drug composition can be to A Erci
Sea silent sick kunming mice brain tissue and spleen have certain protective effect.
The EB contents of measuring brain tissue, are compared with control group, the significantly raised (P of model group Mice brain tissues EB contents
＜ 0.05), illustrate the permeability increase of Blood-Brain Barrier, aggravated exotic matter and entered brain tissue；Compared with model group,
The high, medium and low dosage group Mice brain tissues EB contents of Chinese medicine composition substantially reduce (P ＜ 0.05), illustrate the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe,
Fructus polygoni orientalis Chinese medicine composition is obviously improved the permeability of Blood-Brain Barrier, change and brain, the spleen of brain tissue permeability
Organ index change it is consistent, with the increasing of certain blood-brain barrier permeability while pointing out brain tissue inflammation's significant reaction
Plus.The medical Radix Astragali of motherland described above, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis crude drug composition can be obviously improved Alzheimer disease
The permeability of kunming mice blood-brain barrier.
The content that aβ protein is determined by enzyme-linked immunoassay method is tested, is as a result shown, the hippocampal tissue A Er of model mice
The key link composition aβ protein content of Ci Haimo disease morbidities is significantly raised.The Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis Chinese traditional medicine composition
The high, medium and low dosage of thing is by influenceing aβ protein to develop to improve Alzheimer disease.More than the comprehensive prompting Radix Astragali, when
Return, the radix paeoniae rubrathe, the high, medium and low dosage treatment Alzheimer disease of fructus polygoni orientalis Chinese medicine composition have obvious effect, and pass through A β
To play the therapeutic action to Alzheimer disease.
The content that IL-6, IL-12 albumen are determined by enzyme-linked immunoassay method is tested, as a result shows, is compared with control group, mould
Type group Hippocampus of Mice IL-6, IL-12 protein content is significantly raised (P ＜ 0.05), illustrates model group inflammatory reaction substantially, greatly
The inflammatory factor of amount take part in Alzheimer disease and develop；Compared with model group, the high, medium and low dosage of Chinese medicine composition
Group hippocampal tissue IL-6, IL-12 protein content substantially reduces (P ＜ 0.05), illustrates the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis Chinese medicine
Composition is obviously improved the inflammatory reaction of Hippocampus of Mice, and the trend of IL-6, IL-12 albumen and aβ protein is approached, and illustrates this
A little factors are the important correlation factors that Alzheimer disease is evaluated, while also illustrating in Alzheimer disease Hippocampus of Mice
There is certain correlation with immune inflammatory factor IL-6, IL-12 albumen in key link factors A β albumen.
After experiment terminates, compared with control group, model group kunming mice body weight changes not substantially (P ＞ 0.05)；With model
Group compares, and the high, medium and low dosage group kunming mice Alzheimer disease body weight of Chinese medicine composition changes also unobvious (P ＞ 0.05),
Illustrate that while the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis traditional Chinese medicine composition for treating Alzheimer disease kunming mice will not be changed
Body weight, shows that the Radix Astragali, Radix Angelicae Sinensis, the radix paeoniae rubrathe, fructus polygoni orientalis mixture side effect are smaller, is adapted to carry out controlling for Alzheimer disease
Treat.
In summary, Chinese medicine composition of the invention can improve the learning and memory function of Alzheimer disease model mouse,
Reduce inflammation reaction, reduces the permeability of blood-brain barrier, Chinese medicine composition can by suppress hippocampus A β and other inflammation because
Sub- albumen plays an important role in treatment of alzheimer, while aβ protein is deposited with the change of IL-6, IL-12 protein content
In correlation, specific mechanism needs further experimental study.It is comprehensive above to show Chinese medicine composition pair of the invention
The combined therapy effect of Alzheimer disease is good.
The preferred embodiments of the present invention are the foregoing is only, are merely illustrative for the purpose of the present invention, and it is nonrestrictive；
Those of ordinary skill in the art understand that can carry out many to it in the spirit and scope that the claims in the present invention are limited changes
Become, modification, or even equivalent change, but fall within protection scope of the present invention.
Claims (8)
1. a kind of Chinese medicine composition for being used to treat Alzheimer disease, it is characterised in that by each bulk drug group of following parts by weight
Into：Radix Astragali 30-50 parts by weight, Radix Angelicae Sinensis 4-8 parts by weight, radix paeoniae rubrathe 3-7 parts by weight and fructus polygoni orientalis 15-25 parts by weight.
2. Chinese medicine composition as claimed in claim 1, it is characterised in that be made up of each bulk drug of following parts by weight：The Radix Astragali 40
Parts by weight, the parts by weight of Radix Angelicae Sinensis 6, the parts by weight of the radix paeoniae rubrathe 5 and the parts by weight of fructus polygoni orientalis 20.
3. a kind of clinically suitable Chinese medicine preparation for being used to treat Alzheimer disease, it is characterised in that by claim 1 or 2
The auxiliary material that described Chinese medicine composition is added needed for preparations shaping, is made according to the conventional method for preparing pharmaceutical preparation.
4. Chinese medicine preparation as claimed in claim 3, it is characterised in that described preparation is water decoction, capsule, pill, particle
Agent, tablet or oral liquid.
5. a kind of method of the Chinese medicine composition prepared described in claim 1, it is characterised in that comprise the following steps：
(1) each bulk drug is weighed according to the parts by weight described in claim 1；
(2) mixture of each bulk drug is put into high neck round-bottom flask, adds the distilled water of 7-10 times of medicinal material gross weight, glass
Rod is stirred, and 1.5-2.5h is soaked under enclosed environment, and heating is decocted after 1.5-2.5h, cooling 10min, uses multilayer filtered through gauze
Medicinal material composition, until medicine does not have decoction outflow, obtains first time filtrate；
(3) medicine residue is put into high neck round-bottom flask again, adds crude drug 7-10 times of distilled water of gross weight, repeat step
(2) process carries out second of filtration treatment, obtains second of filtrate, merges filtrate twice；
(4) filtrate obtains the extraction concentrate of Chinese medicine composition after 70 DEG C of evaporation moisture of bitubular steamer, and obtained extraction is dense
Contracting liquid continues to dry 4-5d in 70 DEG C of electric heating constant-temperature blowing drying boxes, and resulting residue is exactly described Chinese medicine composition
Dry cream.
6. method as claimed in claim 5, it is characterised in that also including the dry cream of Chinese medicine composition is placed on into powder in pulverizer
It is broken, it is ground, molecular sieve is handled repeatedly, obtains the pulvis of the Chinese medicine composition.
7. method as claimed in claim 5, it is characterised in that the parts by weight of each bulk drug are：The parts by weight of the Radix Astragali 40, when
Return 6 parts by weight, the parts by weight of the radix paeoniae rubrathe 5 and the parts by weight of fructus polygoni orientalis 20.
8. purposes of the Chinese medicine composition as claimed in claim 1 or 2 in treatment Alzheimer disease drugs are prepared.
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| 治疗老年痴呆病中药应用频次及临床分析;石景洋等;《中国实验方剂学杂志》;20130430;第19卷(第8期);第355-361页，尤其是第356-358页表1，第359页右栏第1段 * |
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