CN109529192A - One kind activating the active device and method of ERK by applying AC field - Google Patents
One kind activating the active device and method of ERK by applying AC field Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61N1/00—Electrotherapy; Circuits therefor
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- A61N1/36—Applying electric currents by contact electrodes alternating or intermittent currents for stimulation
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- C—CHEMISTRY; METALLURGY
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
Abstract
The invention discloses one kind to activate the active device and method of ERK by applying AC field, belongs to biomedical engineering technology field.The equipment includes that at least a pair of metal electrodes, each electrode has conductor and surface；Equipment further includes signal generator and amplifier；Signal generator is connect with amplifier；Amplifier is connect with a pair of metal electrodes；Being generated by control signal generator and amplifier has the active AC field of active cell ERK, and is applied target position with the active AC field of active cell ERK by metal electrode.This method is to apply the AC field of 20V/cm-50V/cm to cell, until the activity of ERK is activated in cell.Present device can activate ERK at any time, so that ERK access is activated, activate the time used in ERK short, it is only necessary to 1s-15min, it is meaningful for preferably simulating the ERK activity under physiological status.
Description
Technical field
The present invention relates to one kind to activate the active device and method of ERK by applying AC field, belongs to biomedical work
Journey technical field.
Background technique
ERK is the english abbreviation of Extracellular Regulated Protein Kinases, refers to extracellular adjusting egg
White kinases, including ERK1 and ERK2 are a member of mitogen-activated protein kinase (MAPK) family, thin for adjusting mammal
Intracellular growth plays an important role.When cell surface tyrosine kinase receptor (such as erbB family, PDGF, FGF and VEGF receptor)
After its ligand activation, the GDP of small G-protein Ras is replaced by GFP and is activated.The activation of Ras leads to a kind of Raf (serine-
Threonine kinase) it is raised to plasma membrane and is activated.The Raf of activation with by MEK1/2 phosphorylation and activating, post activation
MEK1/2 is by ERK1/2 phosphorylation and activates, this access is also referred to as MAPK/ERK access.After ERK Pathway Activation, it will regulate and control many
Cell behavior, including cell Proliferation and differentiation, cellular morphology maintenance and Apoptosis etc..Clinically ERK access and a variety of diseases
Generation it is related to treatment, such as tumour, diabetes and Alzheimer's disease.Therefore, largely small point relevant to ERK activity
Son is invented activity for regulating and controlling ERK access, and the drug of the diseases such as treating cancer is developed with this.ERK signal path is practical
On be by its dynamic change come regulating cell behavior, such as ERK signal path is thin by its temporal dynamic change regulation
Division (Albeck JG1, Mills GB, the Brugge JS.Frequency-modulated pulses of ERK of born of the same parents
activity transmit quantitative proliferation signals.Mol Cell.2013 Jan 24；49
(2): 249-61.) and differentiation (Ryu H, Chung M, Dobrzy ń ski M, Fey D, Blum Y, Sik Lee S, Peter M,
Kholodenko BN,Li Jeon N,Pertz O.Frequency modulation of ERK activation
dynamics rewires cell fate.Mol Syst Biol.2016 Apr 22；12 (4): 866.), and this signal
Access is prevalent in again among various cells.The existing adjusting for ERK pathway activity generallys use inhibitor and inhibits
Or growth factor activates it.The application of inhibitor can cause the lasting inhibition of ERK, and growth factor will lead to ERK sustained activation.
No matter inhibit and activate all to be difficult to control in amplitude, it can not temporally and spatially dynamic regulation ERK signal path
Activity.To keep therapeutic effect unsatisfactory.
Summary of the invention
It is merely able to continue inhibition or activation to solve existing ERK adjusting method, can not dynamically adjust over time and space
The deficiency for controlling ERK signal path, the present invention provides one kind to activate the active device and method of ERK by applying AC field,
The technical solution adopted is as follows:
The purpose of the present invention is to provide one kind to activate the active equipment of ERK by applying AC field, which includes
At least a pair of metal electrodes, each electrode have conductor and surface；The equipment further includes signal generator and amplifier；Institute
Signal generator is stated to connect with amplifier；The amplifier is connect with a pair of metal electrodes；By control signal generator and put
Big device, which generates, has the active AC field of active cell ERK, and being applied by metal electrode to target position has activation thin
The active AC field of born of the same parents ERK.
Preferably, described apply is to execute in vitro.
Preferably, described apply is to execute in vivo.
Preferably, described apply is executed in vitro.
Preferably, applying to target position, which has the active AC field of active cell ERK, is applied to target position
The AC field of 20V/cm-50V/cm, until the activity of ERK is activated in the cell of target position.
Preferably, it is described to target position apply have the active AC field of active cell ERK in the following way it
One:
(1) apply the AC field at least 15min of 20V/cm to cell；
(2) apply the AC field at least 2min of 28V/cm to cell；
(3) apply the AC field at least 1min of 40V/cm to cell；
(4) apply the AC field at least 1s of 50V/cm to cell.
It is highly preferred that described apply to target position has the active AC field of active cell ERK in the following way
One of:
(1) apply the AC field 15min of 20V/cm to cell；
(2) apply the AC field 2min of 28V/cm to cell；
(3) apply the AC field 1min of 40V/cm to cell；
(4) apply the AC field 1s of 50V/cm to cell.
Preferably, the waveform of the AC field is bipolar.
Preferably, the waveform of the AC field is rectangular pulse, and linear change is any in exponential damping and sine wave
The combination of one or more waveforms.
Preferably, the waveform of the AC field is bipolar and is selected from rectangular pulse, linear change, exponential damping and sine
The combination of any one or more waveform in wave.
The present invention also provides one kind to activate the active method of ERK by applying AC field, and this method is applied to cell
Add the AC field of 20V/cm-50V/cm, until the activity of ERK is activated in cell.
Preferably, the cell is isolated cells, such as cell culture.
Preferably, apply any one of the mode of AC field in following manner:
(1) apply the AC field at least 15min of 20V/cm to cell；
(2) apply the AC field at least 2min of 28V/cm to cell；
(3) apply the AC field at least 1min of 40V/cm to cell；
(4) apply the AC field at least 1s of 50V/cm to cell.
It is highly preferred that applying any one of the mode of AC field in following manner:
(1) apply the AC field 15min of 20V/cm to cell；
(2) apply the AC field 2min of 28V/cm to cell；
(3) apply the AC field 1min of 40V/cm to cell；
(4) apply the AC field 1s of 50V/cm to cell.
Preferably, the waveform of the AC field meets one of following condition or meets two following conditions simultaneously:
(1) bipolar；
(2) rectangular pulse, linear change, the combination of any one or more waveform in exponential damping and sine wave.
The object of the present invention is achieved like this:
Electrode is placed in cultured cell in vitro both ends or et al. Ke specific location, switching amplifier is by specific waveform
The AC field of 20V/cm to 50V/cm is applied on cell by electrode, and action time is different because of electric field strength, with electric field
Intensity increases, and the electric field action time of ERK is activated to shorten.It needs persistently to stimulate 15 minutes with 20V/cm electric field-activate ERK；With
28V/cm electric field needs lasting 2 minutes；Lasting 1 minute is needed with 40V/cm electric field；It is only needed 1 second with 50V/cm electric field.Space
On, this equipment can independent regulating cell ERK pathway activity in any position.It, can be by exhausted according to high-frequency alternating current in implementation
Edge separation layer is applied to specific site, it is undesirable that generating electric current in electric field, electrode insulation can be used.Position side
Face both can apply electric field by the way that electrode is implanted into specific position, and can also pass through the non-contact arrangement electrode in surface.
The invention has the advantages that:
The method of current activation ERK can only be continued to inhibit or be activated using inhibitor or activator.And the present invention is set
Standby according to circumstances in specific location, to activate ERK at any time, so that ERK access is dynamically activated, present device swashs
Time used in ERK living is short, it is only necessary to 1s-15min, it is meaningful for preferably simulating the ERK activity under physiological status.
Detailed description of the invention
Fig. 1 is to generate electric field to target area schematic diagram.
Fig. 2 is the schematic diagram for shifting reporter gene ERKTR.
Fig. 3 is that Cell culture invitro applies electric field arrangement structural schematic diagram；A is the top view of electric field action cell, b in figure
For the sectional view of the partial structurtes of electric field action cell, c is to install 1 equipment of embodiment to electric field action cell to apply electric field
Structural schematic diagram, 1 is upper matrix in figure, and 2 be lower substrate, and 3 be electric field action cell compartments, and 4 is accommodate culture medium cell, and 5 are
The gap of water is accommodated, 6 be the equipment of embodiment 1, and 7 be voltmeter, and 8 be a pair of metal electrodes in 1 equipment of embodiment, and 9 be one
To silver silver chloride electrode, 10 be cell.
Fig. 4 is electric field effect in the 15 minutes activation that electric field strength is 20V/cm, frequency 200Hz, waveform are bipolar rectangular wave
ERK；Fig. 4 a shows the change in location of intracellular ERKTR before and after electrical field stimulation in figure；Fig. 4 b is that ERKTR karyoplasmic ratio becomes at any time
Change, intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio, and shade shows standard deviation, number of cells > 100；Fig. 4 c counts electric field thorn
The cumulative mean value of ERKTR nucleocytoplasmic ratio before swashing and after electro photoluminescence, there were significant differences therebetween, p < 0.001 * *.
Fig. 5 is electric field strength 28V/cm, and frequency 20Hz, the electric field continuous action that waveform is bipolar rectangular wave activate ERK；Figure
Middle Fig. 5 a shows the change in location of intracellular ERKTR before and after electrical field stimulation；Fig. 5 b is that ERKTR karyoplasmic ratio changes over time, intermediate
Black curve is the mean value of ERKTR nucleocytoplasmic ratio, and shade shows standard deviation, number of cells > 100.
Fig. 6 is electric field strength 28V/cm, and frequency 50Hz, the electric field continuous action that waveform is bipolar rectangular wave activate ERK；Figure
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio, and shade shows standard deviation, number of cells > 100.
Fig. 7 is electric field strength 28V/cm, frequency 500Hz, 2 minutes activation ERK of electric field action that waveform is bipolar rectangular wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Fig. 8 is electric field strength 28V/cm, frequency 10Hz, 3 minutes activation ERK of electric field action that waveform is bipolar rectangular wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Fig. 9 is electric field strength 28V/cm, frequency 20Hz, 3 minutes activation ERK of electric field action that waveform is bipolar rectangular wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 10 is electric field strength 28V/cm, frequency 50Hz, 3 minutes activation ERK of electric field action that waveform is bipolar rectangular wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 11 is electric field strength 28V/cm, and frequency 150Hz, electric field action 3 minutes that waveform is bipolar rectangular wave are activated
ERK；Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 12 is electric field strength 28V/cm, and frequency 500Hz, electric field action 3 minutes that waveform is bipolar rectangular wave are activated
ERK；Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 13 is electric field strength 28V/cm, and frequency 500Hz, electric field action 3 minutes that waveform is bipolar triangular wave are activated
ERK；Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 14 is electric field strength 40V/cm, frequency 60Hz, 1 minute activation ERK of electric field action that waveform is bipolar sine wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Figure 15 is electric field strength 50V/cm, frequency 60Hz, 1 second activation ERK of electric field action that waveform is bipolar sine wave；
Intermediate black curve is the mean value of ERKTR nucleocytoplasmic ratio in figure, and shade shows standard deviation, number of cells > 100.
Specific embodiment
Below with reference to embodiment, the present invention will be further described, it should be understood that specific embodiment described herein is only
To explain the present invention, it is not intended to limit the present invention.It will be appreciated by the person skilled in the art that can be to this description and accompanying drawings
Described in specific example make a change, these change within scope and spirit of the present invention.Following example is not specified specifically
The experimental method of condition, usually according to the known approaches of this field.
Embodiment 1
It present embodiments provides a kind of pass through and applies the AC field activation active equipment of ERK, as shown in Figure 1, the equipment
Including at least a pair of metal electrodes, each electrode has conductor and surface；Equipment further includes signal generator and amplifier；Institute
Signal generator is stated to connect with amplifier；The amplifier is connect with a pair of metal electrodes；By control signal generator and put
Big device, which generates, has the active AC field of active cell ERK, and being applied by metal electrode to target position has activation thin
The active AC field of born of the same parents ERK.
Embodiment 2
It present embodiments provides a kind of utilization equipment described in embodiment 1 and applies the AC field activation active method of ERK,
It is embodied as follows:
1, cell used and ERK detection method are tested:
Cell used is tested from people's normal mammary epithelial MCF10A cell line.The activity test method of ERK is
Using a kind of displacement reporter gene (ERK Translocation reporter, ERKTR).This displacement reporter gene is stablized
It is transfected into cell.As shown in Fig. 2, when ERK inactivation, ERKTR's is gathered in nucleus the principle of ERKTR, when ERK is activated
When, ERKTR is transferred in extranuclear cytoplasm.ERKTR is connected to red fluorescent protein, and the position of ERKTR is by being taken
Depending on distribution of the red fluorescent protein of band in cell.It can obtain ERK's in real time by the change in location of ERKTR in this way
Activity change.In addition, the position of each nucleus is marked by stable transfection another nuclear location yellow fluorescent protein gene,
Each cell in this way, ERKTR is in nucleus or extranuclear distribution is assured that.The work of the ERK of each cell
Property can the ratio of ERKTR fluorescence intensity be (referred to as in ERKTR fluorescence intensity and nucleus in approximate quantification of cytoplasm
ERKTR karyoplasmic ratio).The detailed process reference literature of ERK detection method: Sparta B, Pargett M, Minguet M,
Distor K,Bell G,Albeck JG.Receptor Level Mechanisms Are Required for
Epidermal Growth Factor(EGF)-stimulated Extracellular Signal-regulated Kinase
(ERK)Activity Pulses.J Biol Chem.2015 Oct 9；290(41):24784-92.
2, Cell culture invitro applies electric field arrangement
The equipment that AC signal generator, switch/amplifier and a pair of metal electrodes 8 are connected into composition embodiment 1 referring to Fig.1
6, then the cell in electric field action cell is stimulated with the equipment 6 of embodiment 1.The structural schematic diagram of electric field action cell
Referring to Fig. 3 a and Fig. 3 b, 1 equipment of embodiment is installed to electric field action cell and applies the structural schematic diagram of electric field referring to Fig. 3 c.
As shown in figure 3, electric field action cell compartments include one layer of upper matrix 1, and lower substrate 2, electric field action cell compartments 3,
It accommodates culture medium cell 4 and accommodates the gap 5 of water；Upper matrix 1 is PDMS and 2 material of lower substrate is 9cm culture dish；Upper matrix 1
Diameter is 6cm, small as electric field action cell with the small strip side's groove of 20mm × 1mm × 0.17mm with a thickness of 1mm
Room 3, strip side groove both ends are the semicircular hole 4 4cm there are two diameter, to accommodate culture medium.Culture dish is stamped 4 circles
Hole, to pass through a pair of metal electrodes 8 and a pair of of silver silver chloride electrode 9.MCF10A cell 10 is seeded in electric field action cell
In.In experimentation, in 5 position Jia Shui of gap, moist environment is kept, silver silver chloride electrode 9 is connected with multimeter 7, for supervising
Electric field in control experiment.
3, electric field-activate ERK
Electrode is placed in the both ends of culture cell device, is situated between by the cell culture medium in culture cell device as conductive
Matter, the AC field for applying 20V/cm to the cell of dispersion in cell culture medium persistently stimulate cell with ERK in active cell
Activity；Comprise the concrete steps that applying the waveform of 20V/cm AC field to electric field action cell is bipolar rectangular wave, frequency is
200Hz, the electric field action time (persistently stimulating the time of cell) is 15 minutes, closes electric field later, as a result as Fig. 4 is shown.
As shown in fig. 4 a, before electrical field stimulation, cell shown in arrow, ERKTR is concentrated in nucleus, after electrical field stimulation,
ERKTR is transferred in cytoplasm, and ERK is by electric field-activate.Fig. 4 b shows, (electric field started at 0 minute), ERKTR after electric field action
Nucleocytoplasmic ratio reduces, and after electric field action 15 minutes, the average value of ERKTR karyoplasmic ratio increases.There is aobvious ERKTR karyoplasmic ratio before and after electro photoluminescence
Sex differernce is write, referring to fig. 4 c, P < 0.001.
Embodiment 3
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 20Hz, electric field continuous action time are 60 minutes.
As a result as Fig. 5 a shows that before electrical field stimulation, cell shown in arrow, ERKTR is concentrated in nucleus, electrical field stimulation
Afterwards, ERKTR is transferred in cytoplasm, and ERK is by electric field-activate.Fig. 5 b is shown, after electric field action (electric field started at 0 minute),
The average value of ERKTR karyoplasmic ratio first reduces, significantly raised later.
Embodiment 4
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 500Hz, electric field continuous action time are 60 minutes.As a result as Fig. 6 shows that (electric field is at 0 point after electric field action
Clock starts), the average value of ERKTR karyoplasmic ratio first reduces, and significantly raised later, ERK is by electric field-activate.
Embodiment 5
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 500Hz, electric field action time are 2 minutes, close electric field later.As a result such as Fig. 7 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
Embodiment 6
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 10Hz, electric field action time are 3 minutes, close electric field later, and as a result such as Fig. 8 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
Embodiment 7
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 20Hz, electric field action time are 3 minutes, close electric field later, and as a result such as Fig. 9 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
Embodiment 8
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 50Hz, electric field action time are 3 minutes, close electric field later, and as a result such as Figure 10 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
Embodiment 9
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 150Hz, electric field action time are 3 minutes, close electric field later, as a result such as Figure 11 is shown, after electric field action
(electric field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, significantly raised later, then gradually decreases.Show ERK quilt
Electric field-activate returns to original state again later.
Embodiment 10
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar square
Shape wave, frequency 500Hz, electric field action time are 3 minutes, close electric field later, as a result such as Figure 12 is shown, after electric field action
(electric field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, significantly raised later, then gradually decreases.Show ERK quilt
Electric field-activate returns to original state again later.
Embodiment 11
The present embodiment the difference from example 2 is that: to electric field action cell apply 28V/cm electric field, waveform be bipolar three
Angle wave, frequency 500Hz, electric field action time are 3 minutes, close electric field later, as a result such as Figure 13 is shown, after electric field action
(electric field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, significantly raised later, then gradually decreases.Show ERK quilt
Electric field-activate returns to original state again later.
Embodiment 12
The present embodiment the difference from example 2 is that: to electric field action cell apply 40V/cm electric field, waveform be it is bipolar just
String wave, frequency 60Hz, electric field action time are 1 minute, close electric field later, and as a result such as Figure 14 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
Embodiment 13
The present embodiment the difference from example 2 is that: to electric field action cell apply 50V/cm electric field, waveform be it is bipolar just
String wave, frequency 60Hz, electric field action time are 1 second, close electric field later, and as a result such as Figure 15 is shown, (electricity after electric field action
Field started at 0 minute), the average value of ERKTR karyoplasmic ratio first reduces, and it is significantly raised later, then gradually decrease.Show that ERK is electric
Field activation returns to original state again later.
It is above-mentioned it is demonstrated experimentally that in the range of 20V/cm to 50V/cm, electric field action 1 second to 15 minutes, respectively can be with
The activity of active cell ERK.Although the present invention has been disclosed in the preferred embodiment as above, it is not intended to limit the invention,
Any person skilled in the art can do various changes and modification, therefore this without departing from the spirit and scope of the present invention
The protection scope of invention should subject to the definition of the claims.
Claims (10)
1. one kind activates the active equipment of ERK by applying AC field, which is characterized in that the equipment includes at least a pair of of gold
Belong to electrode, each electrode has conductor and surface；The equipment further includes signal generator and amplifier；The signal occurs
Device is connect with amplifier；The amplifier is connect with a pair of metal electrodes；Tool is generated by control signal generator and amplifier
There is the active AC field of active cell ERK, and applying by metal electrode to target position has active cell ERK active
AC field.
2. equipment according to claim 1, which is characterized in that it is described application be execute in vitro, or execute in vivo, or
It is executed in vitro.
3. equipment according to claim 1, which is characterized in that applying to target position has active cell ERK active
AC field is to apply the AC field of 20V/cm-50V/cm to target position, up to the activity of ERK in the cell of target position
It is activated.
4. equipment according to claim 3, which is characterized in that described apply to target position has active cell ERK living
The AC field of property is one of in the following way:
(1) apply the AC field at least 15min of 20V/cm to cell；
(2) apply the AC field at least 2min of 28V/cm to cell；
(3) apply the AC field at least 1min of 40V/cm to cell；
(4) apply the AC field at least 1s of 50V/cm to cell.
5. equipment according to claim 3, which is characterized in that described apply to target position has active cell ERK living
The AC field of property is one of in the following way:
(1) apply the AC field 15min of 20V/cm to cell；
(2) apply the AC field 2min of 28V/cm to cell；
(3) apply the AC field 1min of 40V/cm to cell；
(4) apply the AC field 1s of 50V/cm to cell.
6. equipment described in -5 any one claims according to claim 1, which is characterized in that the waveform of the AC field
Meet one of following condition or meet two following conditions simultaneously:
(1) bipolar；
(2) rectangular pulse, linear change, the combination of any one or more waveform in exponential damping and sine wave.
7. one kind activates the active method of ERK by applying AC field, which is characterized in that apply 20V/cm-50V/ to cell
The AC field of cm, until the activity of ERK is activated in cell.
8. the method according to claim 1, wherein applying the mode of AC field appointing in following manner
It anticipates one kind:
(1) apply the AC field at least 15min of 20V/cm to cell；
(2) apply the AC field at least 2min of 28V/cm to cell；
(3) apply the AC field at least 1min of 40V/cm to cell；
(4) apply the AC field at least 1s of 50V/cm to cell.
9. the method according to claim 1, wherein applying the mode of AC field appointing in following manner
It anticipates one kind:
(1) apply the AC field 15min of 20V/cm to cell；
(2) apply the AC field 2min of 28V/cm to cell；
(3) apply the AC field 1min of 40V/cm to cell；
(4) apply the AC field 1s of 50V/cm to cell.
10. according to method described in claim 7-9 any one claim, which is characterized in that the wave of the AC field
Shape meets one of following condition or meets two following conditions simultaneously:
(1) bipolar；
(2) rectangular pulse, linear change, the combination of any one or more waveform in exponential damping and sine wave.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050059153A1 (en) * | 2003-01-22 | 2005-03-17 | George Frank R. | Electromagnetic activation of gene expression and cell growth |
| US20200140845A1 (en) * | 2018-11-06 | 2020-05-07 | Arizona Board Of Regents On Behalf Of Arizona State University | Cell signaling pathway activation by local ac electric field |
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2018
- 2018-11-15 CN CN201811359991.7A patent/CN109529192A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050059153A1 (en) * | 2003-01-22 | 2005-03-17 | George Frank R. | Electromagnetic activation of gene expression and cell growth |
| US20200140845A1 (en) * | 2018-11-06 | 2020-05-07 | Arizona Board Of Regents On Behalf Of Arizona State University | Cell signaling pathway activation by local ac electric field |
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| GUO LIANG: "Controlling ERK Activation Dynamics in Mammary Epithelial Cells with Alternating Electric Fields through Microelectrodes", 《NANO LETTERS》 * |
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