CN104706665B - One kind WS2Nanometer sheet suppresses the method for A beta peptide aggregations and the method for the established A betas aggregation of depolymerization - Google Patents
One kind WS2Nanometer sheet suppresses the method for A beta peptide aggregations and the method for the established A betas aggregation of depolymerization Download PDFInfo
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Abstract
The present invention provides one kind WS2Nanometer sheet suppresses the method for A beta peptide aggregations and the method for the established A betas aggregation of depolymerization, belongs to pharmaceutical technology field.This method suppresses A β aggregation, WS using transition metal tungsten disulfide two-dimension nano materials2Nanometer sheet can adsorb A beta monomers by the Van der Waals force between its planar substrates and aromatic amino acid, meanwhile, WS2Surface is negatively charged, can strengthen A beta monomers and WS with electrostatical binding A β cation cluster region2Combination between lamellar structure, suppress A beta peptide aggregations；The present invention also provides one kind WS2The method of the established A betas aggregation of nanometer sheet depolymerization, WS2Nanometer sheet has obvious photo-thermal effect, being capable of the effective established A betas aggregation of photo-thermal depolymerization under near infrared light；Meanwhile WS2Nanometer sheet can reduce cytotoxicity caused by A β.
Description
Technical field
The invention belongs to pharmaceutical technology field, and in particular to one kind WS2Nanometer sheet suppresses the method for A beta peptide aggregations and depolymerization
The method of established A betas aggregation.
Background technology
Alzheimer disease (AD) is one of most commonly seen form of senile dementia.The clinical symptoms of AD patient mainly show
For cognition dysfunction, gradual memory decline, abnormality of personality etc..Its pathological characteristics be intracerebral extracellular senile plaque expelling deposition with
And cellular neurofibrillary tangles.The main component of wherein senile plaque expelling is beta amyloid peptide (amyloid β-peptide, A β).
The experimental results show A β excess generation and aggregation is AD key pathological event.Suppress A β Assembling Behavior and depolymerization
Established A betas aggregation is prevention and the effective way for treating AD.A beta peptide aggregations process can be regulated and controled and reduce A β toxicity
Material be treatment the very promising drug candidates of AD.
Based on this, substantial amounts of peptide derivative, organic molecule and some metal complexs are designed synthesis and are used to press down
The A β processed established A betas of aggregation or depolymerization.However, because its targeting is poor, blood-brain barrier (BBB) permeability is low and malicious
The features such as side effect is higher, most inhibitor only show medium inhibition and faint depolymerization ability.Therefore compel
New compound or material will be designed for suppressing A beta peptide aggregations and depolymerization A beta aggregations by being essential.In recent years, nano material
Because the property such as its larger specific surface area and unique light, electricity, magnetics and catalytic activity is increasingly subject to the extensive concern of people.Receive
Rice biology blends nanosecond science and technology and life science, turns into the focus of modern science and technology research, in biomedical, electronics
The numerous areas such as, materialogy, environmental science have a good application prospect.Numerous studies show that nano material can pass through
Hydrophobic interaction, pi-pi accumulation effect, Van der Waals force and electrostatic interaction etc. and interaction of biomacromolecules.Nanometer material
Material can treat AD as a kind of new pharmaceutical agent for regulating and controlling A β Assembling Behavior.Have now been found that a variety of nanometers of materials
Material can suppress A β aggregation (Adv.Mater.2013,25,3780-3801.), but these nano materials mainly pass through it
The electric charge and group on surface realizes the regulating and controlling effect to A beta peptide aggregation behaviors.
The content of the invention
The invention aims to provide one kind WS2Nanometer sheet suppresses the method and the established A β of depolymerization of A beta peptide aggregations
The method of fiber aggregate, this method can greatly reduce the toxic side effect of medicine and improve selectivity.
Present invention firstly provides one kind WS2The method that nanometer sheet suppresses A beta peptide aggregations, this method include：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then by the A beta monomers sample and WS after processing2Nanometer sheet is buffering
In solution, it is incubated at 37 DEG C and suppresses A beta peptide aggregations.
Preferably, the method that is handled A beta monomers samples is：A β powder is dissolved in hexafluoroisopropanol
In, 2-6 hour of concussion at 4 DEG C is placed in, hexafluoroisopropanol is dried up with nitrogen stream, is then dissolved in cushioning liquid, at 4 DEG C
Middle standing 6-12 hours.
Preferably, described cushioning liquid is 10mM HEPES, 150mM NaCl, pH 7.3.
The present invention also provides one kind WS2The method of the established A betas aggregation of nanometer sheet depolymerization, this method include：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then the A beta monomers samples after processing are dissolved in buffer solution, 37
It is incubated 7 days at DEG C, obtains A beta aggregations；
Step 3：By WS2Nanometer sheet and A betas aggregation are after 37 DEG C are incubated 20 minutes, near-infrared laser illumination
5min, depolymerization is carried out to established A betas aggregation.
Preferably, the method that is handled A beta monomers samples is：A β powder is dissolved in hexafluoroisopropanol
In, 2-6 hour of concussion at 4 DEG C is placed in, hexafluoroisopropanol is dried up with nitrogen stream, is then dissolved in cushioning liquid, at 4 DEG C
Middle standing 6-12 hours.
Preferably, described cushioning liquid is 10mM HEPES, 150mM NaCl, pH 7.3.
Beneficial effects of the present invention
Present invention firstly provides one kind WS2The method that nanometer sheet suppresses A beta peptide aggregations, this method utilize the sulphur of transition metal two
Change tungsten (WS2) two-dimension nano materials suppress A β aggregation, WS2Nanometer sheet can by its planar substrates and aromatic amino acid it
Between Van der Waals force absorption A beta monomers, meanwhile, WS2Surface is negatively charged, can strengthen A β with electrostatical binding A β cation cluster region
Monomer and WS2Combination between lamellar structure, suppress A beta peptide aggregations；Test result indicates that：WS2Nanometer sheet can significantly suppress A β and gather
Collection, and its inhibition is in concentration dependent；WS2To the inhibitory action of A beta monomers in complex system such as bovine serum albumin(BSA)
(BSA) remain able to realize in solution and mouse cerebrospinal fluid (CSF).
The present invention also provides one kind WS2The method of the established A betas aggregation of nanometer sheet depolymerization, WS2Nanometer sheet exists
There is higher optical absorption near infrared region, using the property of its near infrared absorption, effectively can melt cancer cell simultaneously by photo-thermal
Reduce the tumor size of mouse, based on this, this method utilizes WS2Simultaneously the established A betas of photo-thermal depolymerization gather nano-lamellar structure
Collective, compared with traditional chemotherapy and radiation therapy, the advantage of thermotherapy is the diseased region just meeting heat production of only illumination,
And other normal structures are unaffected, can greatly reduce the toxic side effect of medicine and improve selectivity.Experimental result table
It is bright：WS2Nanometer sheet has obvious photo-thermal effect, under near infrared light can effectively the established A β of photo-thermal depolymerization it is fine
Tie up aggregation；Meanwhile WS2Nanometer sheet can reduce cytotoxicity caused by A β；WS2Photo-thermal depolymerization to A beta aggregations is made
Remain able to realize in complex system such as bovine serum albumin(BSA) (BSA) solution and mouse cerebrospinal fluid (CSF).
Brief description of the drawings
Fig. 1 is the WS of the step 1 of embodiment 1 synthesis2Nanometer sheet atom is tried hard to；
Fig. 2 is embodiment 1WS2The inhibitory action ThT fluoroscopic examination figures that nanometer sheet is assembled to A β 1-40；
Fig. 3 is embodiment 1A β 1-40 and WS2Nanometer sheet be incubated altogether 7 days after decay ATR-FTIR collection of illustrative plates；
Fig. 4 is embodiment 1A β 1-40 and WS2Nanometer sheet be incubated altogether 7 days after CD spectrograms；
Fig. 5 is embodiment 1WS2Nanometer sheet assembles the AFM shape appearance figures influenceed to A β 1-40；
Fig. 6 is embodiment 2WS2Photo-thermal depolymerisation ThT fluoroscopic examination figure of the nanometer sheet to A β 1-40 fiber aggregates；
Fig. 7 is embodiment 2WS2The CD spectrograms of photo-thermal depolymerisation of the nanometer sheet to A β 1-40 fiber aggregates；
Fig. 8 is embodiment 2WS2Nanometer sheet assembles the AFM shape appearance figures influenceed to A β 1-40；
Fig. 9 is embodiment 3WS2The inhibitory action ThT fluoroscopic examination figures that nanometer sheet is assembled to A β 1-40；
Figure 10 is embodiment 4WS2Photo-thermal depolymerisation ThT fluoroscopic examination figure of the nanometer sheet to A β 1-40 fiber aggregates；
Figure 11 mtt assay detects WS2Nanometer sheet suppresses figure to cytotoxicity caused by A β 1-40.
Embodiment
Present invention firstly provides one kind WS2The method that nanometer sheet suppresses A beta peptide aggregations, this method include：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then by the A beta monomers sample and WS after processing2Nanometer sheet is buffering
In solution, it is incubated at 37 DEG C and suppresses A beta peptide aggregations.
According to the present invention, WS is prepared first2Nanometer sheet, WS2The preparation method of nanometer sheet is technical side commonly used in the art
Method, it is not particularly limited, is preferably：In a nitrogen environment, by WS2Powder dissolves in a solvent, and described solvent is normal-butyl
Lithium solution or hexane solution, after 2 days, mixture is filtered with filter paper, and washed with hexane, described filter paper is preferably Whatman#
41 filter paper, will add deionized water in obtained leather hard sediment, be ultrasonically treated 1 hour, then by solution centrifugal, spend from
Sub- 3 removal lithium ions of water washing and undispersed bulky grain, collect supernatant, are dialysed 5 days with deionized water, obtain WS2Receive
Rice piece.Resulting WS2The average thickness of nanometer sheet is 1.7nm, average diameter 200nm.
According to the present invention, the method that is handled A beta monomers samples is preferably：A β powder is dissolved in hexafluoro
In isopropanol, 2-6 hour of concussion at 4 DEG C is placed in, it is all dissolved, storing solution is preserved at -20 DEG C, is using it
Before, hexafluoroisopropanol is dried up with nitrogen stream, is then dissolved in cushioning liquid, 6-12 hours are stood in 4 DEG C.Described is slow
It is preferably 10mM HEPES, 150mM NaCl, pH 7.3 to rush solution.
According to the present invention, by the A beta monomers sample and the WS of various concentrations after processing2Nanometer sheet is in cushioning liquid, 37
DEG C be incubated suppress A beta peptide aggregations.The concentration of described A beta monomers samples is 50 μM, WS2The concentration of nanometer sheet is 0-100 μ gmL-1, institute
The cushioning liquid stated is preferably 10mM HEPES, 150mM NaCl, pH 7.3.WS2Sulphur is passed through to the rejection ability of A beta peptide aggregations
Plain T (ThT) fluorescence spectrum, circular dichroism spectra (CD), atomic force (AFM) and decay In situ ATR-FTIR (ATR-FTIR) means
Characterize.
WS2Nano-material surface combination A beta monomers, the concentration of monomer in solution can be reduced, and A β fibrosis and monomer are dense
Spend it is closely related, reduce monomer concentration can suppress A beta peptide aggregations.Meanwhile A beta monomers form oligomer by intermolecular interaction
Process be reversible.The reduction of monomer concentration can cause the reaction to be carried out to the direction of oligomer depolymerization, reduce and deposited in solution
Aggregation core or seed content, so as to effectively suppressing its accumulation process.WS2Nanoscale twins can pass through Van der Waals force
A beta monomers are adsorbed with electrostatic interaction, so as to reach the effect for suppressing A beta peptide aggregations.
The present invention also provides one kind WS2The method of the established A betas aggregation of nanometer sheet depolymerization, this method include：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then the A beta monomers samples after processing are dissolved in buffer solution, 37
It is incubated 7 days at DEG C, obtains A beta aggregations；
Step 3：By WS2Nanometer sheet and A betas aggregation are after 37 DEG C are incubated 20 minutes, near-infrared laser illumination
5min, depolymerization is carried out to established A betas aggregation.
According to the present invention, WS is prepared first2Nanometer sheet, WS2The preparation method of nanometer sheet is technical side commonly used in the art
Method, it is not particularly limited, is preferably：In a nitrogen environment, by WS2Powder dissolves in a solvent, and described solvent is normal-butyl
Lithium solution or hexane solution, after 2 days, mixture is filtered with filter paper, and washed with hexane, described filter paper is preferably Whatman#
41 filter paper, will add deionized water in obtained leather hard sediment, be ultrasonically treated 1 hour, then by solution centrifugal, spend from
Sub- 3 removal lithium ions of water washing and undispersed bulky grain, collect supernatant, are dialysed 5 days with deionized water, obtain WS2Receive
Rice piece.Resulting WS2The average thickness of nanometer sheet is 1.6nm, average diameter 200nm.
According to the present invention, the method that is handled A beta monomers samples is preferably：A β powder is dissolved in hexafluoro
In isopropanol, 2-6 hour of concussion at 4 DEG C is placed in, it is all dissolved, storing solution is preserved at -20 DEG C, is using it
Before, hexafluoroisopropanol is dried up with nitrogen stream, is then dissolved in cushioning liquid, 6-12 hours are stood in 4 DEG C.Described is slow
It is preferably 10mM HEPES, 150mM NaCl, pH 7.3 to rush solution.
According to the present invention, the A beta monomers samples after above-mentioned processing are dissolved in buffer solution, is incubated 7 days, obtains at 37 DEG C
A beta aggregations；Described cushioning liquid is preferably 10mM HEPES, 150mM NaCl, pH 7.3.By WS2Nanometer sheet and A
Beta aggregation near-infrared laser illumination 5min, solves after 37 DEG C are incubated 20 minutes to established A betas aggregation
It is poly-.Described near-infrared laser intensity is 1 Wcm-2；WS2Thioflavin T is passed through to the effect of photo-thermal depolymerization A beta aggregations
(ThT) fluorescence spectrum, circular dichroism spectra (CD), atomic force (AFM) and decay In situ ATR-FTIR (ATR-FTIR) means table
Sign, the mitigation to cytotoxicity caused by A beta peptide aggregation bodies pass through MTT methods and membranolysis simulated experiment explanation.
With reference to embodiment, the present invention will be further described in detail.
Embodiment 1
WS2Nanometer sheet suppresses A β 1-40 self aggregation：
Step 1：Prepare WS2Nanometer sheet：In a nitrogen environment, 100mg WS2It is molten that powder is dissolved in 1.6 M n-BuLis
Reacted in liquid, after 2 days, mixture is filtered with Whatman#41 filter paper, and washed 3 times with 100mL hexanes, it is half-dried what is obtained
300mL deionized waters are added in shape sediment, is ultrasonically treated 1 hour, then by solution centrifugal, is washed with deionized 3 times and goes
Except lithium ion and undispersed bulky grain, supernatant is collected, is dialysed 5 days with deionized water, obtains WS2Nanometer sheet；
Step 2：First by A β 1-40 (kinds of goods U10012, being bought from American Peptide) with the dense of 1mg/ml
Degree is dissolved in hexafluoroisopropanol, and bottleneck is sealed, and is placed in 2 hours of vibration at 4 DEG C, is promoted it all to dissolve, storing solution is in -20
Preserve at DEG C, before the use, dried up hexafluoroisopropanol with soft nitrogen stream, be then re-dissolved in buffer accordingly it is molten
In liquid (10mM HEPES, 150mM NaCl, pH 7.3), balance 6 hours are stood in 4 DEG C of refrigerators；By 50 μM of A β after processing
1-40 and various concentrations (0-100 μ gmL-1) WS2Nanometer sheet is in cushioning liquid, in 37 DEG C of incubations, ThT fluorescence detection methods
Analyze WS2The influence that nanometer sheet is assembled to A β 1-40.
Fig. 1 is the WS of the step 1 of embodiment 1 synthesis2Nanometer sheet atom is tried hard to, wherein figure A tries hard to for atom, figure B is corresponding
Height map, it will be seen from figure 1 that synthesis WS2Nanometer sheet average thickness is 1.7nm, average diameter 200nm.
Fig. 2 is embodiment 1WS2The inhibitory action ThT fluoroscopic examination figures that nanometer sheet is assembled to A β 1-40；Fig. 2 illustrates A β 1-
40 from the ThT Fluorescence Increasings curve of collecting process and incubation time be in typical S types, shows that it meets nucleation-dependence and assembles mould
Type, and WS2Nanometer sheet then suppresses A β 1-40 fibrotic processes in concentration dependent.
Fig. 3 is embodiment 1A β 1-40 and WS2Nanometer sheet be incubated altogether 7 days after decay ATR-FTIR collection of illustrative plates；Fig. 4 is real
Apply 1A β a 1-40 and WS2Nanometer sheet be incubated altogether 7 days after CD spectrograms, after Fig. 3 and Fig. 4 show A β 1-40 via aging is incubated
Beta sheet structure can be changed into.And and WS2After being incubated altogether, A β 1-40 are still random coil structure.
Fig. 5 is embodiment 1WS2Nanometer sheet assembles the AFM shape appearance figures influenceed to A β 1-40, wherein figure A is A β 1-40's
AFM shape appearance figures, figure B is A β 1-40 and WS2Mixture AFM shape appearance figures, Fig. 5 shows A β 1-40 WS2After nanometer sheet effect,
Only a small amount of random precipitation produces, and is not substantially observed aggregate structure, illustrate WS2Nanometer sheet can effectively press down
The formation of A β 1-40 beta sheet structures processed.
Embodiment 2
WS2The established A β 1-40 fiber aggregates of nanometer sheet photo-thermal depolymerization：
Step 1：Prepare WS2Nanometer sheet：In a nitrogen environment, 100mg WS2It is molten that powder is dissolved in 1.6 M n-BuLis
Reacted in liquid, after 2 days, mixture is filtered with Whatman#41 filter paper, and washed 3 times with 100mL hexanes, it is half-dried what is obtained
300mL deionized waters are added in shape sediment, is ultrasonically treated 1 hour, then by solution centrifugal, is washed with deionized 3 times and goes
Except lithium ion and undispersed bulky grain, supernatant is collected, is dialysed 5 days with deionized water, obtains WS2Nanometer sheet；
Step 2：First by A β 1-40 (kinds of goods U10012, being bought from American Peptide) with the dense of 1mg/ml
Degree is dissolved in hexafluoroisopropanol, and bottleneck is sealed, and is placed in 2 hours of vibration at 4 DEG C, is promoted it all to dissolve, storing solution is in -20
Preserve at DEG C, before the use, dried up hexafluoroisopropanol with soft nitrogen stream, be then re-dissolved in buffer accordingly it is molten
In liquid (10mM HEPES, 150mM NaCl, pH 7.3), balance 6 hours are stood in 4 DEG C of refrigerators；By the A β after above-mentioned processing
1-40 samples are dissolved in buffer solution (10 mM HEPES, 150mM NaCl, pH 7.3), are incubated 7 days at 37 DEG C, are obtained A β 1-
40 fiber aggregates；
Step 3：By 40 μ gmL-1WS2Nanometer sheet and 50 μM of A β 1-40 fiber aggregates buffer solution (10 mM HEPES,
150mM NaCl, pH 7.3) in, after 37 DEG C are incubated 20 minutes, make WS2The Targeting Effect of A β 1-40 fibers is maximized, used
1Wcm-2The laser of power density is irradiated 5 minutes, and depolymerization is carried out to established A β 1-40 fiber aggregates.
Fig. 6 is embodiment 2WS2Photo-thermal depolymerisation ThT fluoroscopic examination figure of the nanometer sheet to A β 1-40 fiber aggregates；Figure
7 be embodiment 2WS2The CD spectrograms of photo-thermal depolymerisation of the nanometer sheet to A β 1-40 fiber aggregates；Fig. 8 is embodiment 2WS2
Nanometer sheet assembles the AFM shape appearance figures influenceed to A β 1-40, wherein figure A is in WS2In the presence of nanometer sheet, A β 1-40 fiber aggregates
AFM shape appearance figures, figure B be near infrared light photo-irradiation treatment after A β 1-40 fiber aggregates and WS2After mixture, A β 1-40 AFM
Shape appearance figure；Fig. 6-8 can illustrate that β-lamellar structure in A β 1-40 drops to 22.4% from 54.8% before, and single
Near-infrared irradiates or WS2Itself all it is unable to the structure of the established A β 1-40 fiber aggregates of depolymerization.
Embodiment 3
WS2Nanometer sheet suppresses A β 1-40 aggregations in cerebrospinal fluid
Reaction condition and step are with embodiment 1, and difference is, by A the β 1-40 and WS after processing2Nanometer sheet is in brain
It is incubated in spinal fluid.
Fig. 9 is embodiment 3WS2The inhibitory action ThT fluoroscopic examination figures that nanometer sheet is assembled to A β 1-40, Fig. 9 can be seen
Go out, in cerebrospinal fluid, WS2Nano flake remains to suppress A β 1-40 fibrotic processes in concentration dependent.
Embodiment 4
WS2Nanometer sheet photo-thermal depolymerization A β 1-40 fiber aggregates in cerebrospinal fluid
Reaction condition and step are with embodiment 2, and difference is, by WS2Nanometer sheet exists with A β 1-40 fiber aggregates
It is incubated in cerebrospinal fluid.
Figure 10 is embodiment 4WS2Nanometer sheet to the photo-thermal depolymerisation ThT fluoroscopic examination figures of A β 1-40 fiber aggregates,
Figure 10 can be seen that compared with A β 1-40 fiber aggregates, and after the treatment with irradiation of near infrared light, fluorescence intensity drops to
58%.
Embodiment 5
WS2Nanometer sheet alleviates cytotoxicity experiment caused by A β 1-40
PC12 cells (mouse pheochromocytoma) are by American Type Culture Collecti (American Type Culture
Collection) provide.IMDM culture mediums (Gibco BRL) equipped with 5% hyclone and 10% horse serum are used for cell
Cultivate, the environment of moistening, 37 DEG C of constant temperature are kept in incubator, and pass to containing 5%CO2Air.PC12 cells are thin with 10000
The hole density of born of the same parents is placed on 96 orifice plates, and is incubated in incubator 24 hours and is allowed cell attachment and propagation.
Inhibition test：
50 μM of A β 1-40 first with various concentrations WS2Nanometer sheet (20 μ gmL-1, 40 μ gmL-1) piece is in vitro in cushioning liquid
37 DEG C are incubated 7 days.A β 1-40 sample dispersions are continued into PC12 culture medium to cultivate 36 hours (A β 1-40 final concentration of 5 μ
M, WS2The final concentration of nanometer sheet is respectively 2 μ gmL-1, 4 μ gmL-1)。
Depolymerization is tested：The WS of cell culture medium various concentrations2(final concentration of 2 μ gmL-1, 4 μ gmL-1) replace, and rapidly
Established A β 1-40 aggregations (final concentration of 5 μM) are added in cell.Then, orifice plate 808nm near infrared lasers
With 0.5Wcm-2Power density irradiate 5 minutes.Test as a comparison, while WS will be contained2- A β 1-40 PC12 cells are placed
In dark place.Cell continues after being incubated 36 hours, adds final concentration of 0.5mgmL-1MTT continue culture 4 hours.Remove culture
Dissolved after base with DMSO, determine the absorption value under 570nm and 630nm wavelength.
Figure 11 mtt assay detects WS2Nanometer sheet suppresses figure, wherein * P to cytotoxicity caused by A β 1-40< 0.05,**P<
0.01,***P<0.001., Figure 11 is shown, A β 1-40 fibril aggregation physical efficiencys cause cytoactive to decline 44%.In Inhibition test,
WS2A β toxicity can be significantly reduced by being incubated altogether with A β 1-40, and reduction and the WS of its toxicity2In concentration dependent.The opposing party
Face, the cell handled with A β 1-40 fiber aggregates is in WS2Effect is lower to use near infrared light 5 minutes, and the survival rate of cell exists
Increase to 88% in experimental error, independent WS2Effect or laser irradiation have no significant effect to the survival rate of cell.
Claims (4)
1. one kind WS2The method that nanometer sheet suppresses A beta peptide aggregations, it is characterised in that this method includes：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then by the A beta monomers sample and WS after processing2Nanometer sheet is in mouse brain ridge
In liquid, it is incubated at 37 DEG C and suppresses A beta peptide aggregations.
2. one kind WS according to claim 12The method that nanometer sheet suppresses A beta peptide aggregations, it is characterised in that described to A β
The method that monomer sample is handled is：A β powder is dissolved in hexafluoroisopropanol, 2-6 hour of concussion at 4 DEG C is placed in, uses
Nitrogen stream dries up hexafluoroisopropanol, is then dissolved in cushioning liquid, and 6-12 hours are stood in 4 DEG C.
3. one kind WS2The method of the established A betas aggregation of nanometer sheet depolymerization, it is characterised in that this method includes：
Step 1：Prepare WS2Nanometer sheet；
Step 2：A beta monomers samples are handled, then the A beta monomers samples after processing are dissolved in mouse cerebrospinal fluid, 37
It is incubated 7 days at DEG C, obtains A beta aggregations；
Step 3：By WS2Nanometer sheet and A betas aggregation are after 37 DEG C are incubated 20 minutes, near-infrared laser illumination 5min, to
The A betas aggregation of formation carries out depolymerization.
4. one kind WS according to claim 32The method of the established A betas aggregation of nanometer sheet depolymerization, its feature
It is, the method that is handled A beta monomers samples is：A β powder is dissolved in hexafluoroisopropanol, is placed at 4 DEG C
2-6 hour is shaken, hexafluoroisopropanol is dried up with nitrogen stream, is then dissolved in cushioning liquid, it is small that 6-12 is stood in 4 DEG C
When.
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| CN106115786B (en) * | 2016-06-27 | 2017-09-15 | 中国地质大学（北京） | Tungsten disulfide nanosheet tubular aggregate and preparation method thereof |
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