CN110090300A - A kind of method and its application for the Regeneration and Repair promoting histoorgan - Google Patents
A kind of method and its application for the Regeneration and Repair promoting histoorgan Download PDFInfo
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- CN110090300A CN110090300A CN201810082885.2A CN201810082885A CN110090300A CN 110090300 A CN110090300 A CN 110090300A CN 201810082885 A CN201810082885 A CN 201810082885A CN 110090300 A CN110090300 A CN 110090300A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
Abstract
The present invention relates to a kind of method and its application of Regeneration and Repair for promoting histoorgan.The inventors discovered that Myosin inhibitor can destroy cell machinery stress system stable state, cytogenetics repair ability is greatly improved using stress reaction similar to stress reaction and regenerative response during regeneration in excitation.The Myosin inhibitor is preferably (-)-Blebbistatin.Method of the invention only needs single small molecule or single factor test to handle, easy to operate, reproducible, can be used as excitation stress reaction, excitation regenerative response, a kind of new method for improving cytogenetics repair ability and/or promoting the Regeneration and Repair of histoorgan.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to it is a kind of promote histoorgan Regeneration and Repair method and its
Related application.
Background technique
Multicellular organism body tissue, organ are made of the extracellular matrix of parenchyma and interstitial cell and its secretion.It is real
Cell plastid refers to tissue, (parenchyma of such as brain is exactly neuron, liver parenchyma to the main structure and function cell of organ
Cell is liver cell), interstitial cell and extracellular matrix constitute tissue and the stromal portion of organ and (mainly have interstitial cell, glue
Former albumen, Laminin lens, fibronectin, elastin laminin, proteoglycans, glycoprotein, glycosaminoglycan), it is main to play mechanical branch
Support and connection function, extracellular matrix constitute and maintain the microenvironment of cellular physiological events, are to carry out letter between cell and cell
Number conduction bridge, participate in and regulate and control a variety of pathological processes, risen in tissue trauma reparation, regeneration and fibrotic processes
Important function.
During regeneration, damaging, which causes one stress rapidly send out, is answered, a series of stress proteins such as heat shock
Protein family expression up-regulation, stress reaction occur in damage 24 hours；The 2-2.5 days initiation renewable hairs after impaired are answered,
Some regeneration starting related genes and the gene upregulation for organizing specialization；Go out in the histocyte of the regeneration specific differentiation of final stage
It is existing, regenerate new tissue.
The reason of evolution, mammal have very limited power of regeneration.The tissue cell insult of higher mammal
After can cause histocyte that denaturation, necrosis and with inflammatory reaction occurs.If damaging very little, the histocyte periphery of damage
Proliferation for repairing will occur for normal parenchyma, to restore normal institutional framework and function completely.However if compared with
Macrolesion or repeatedly damage be when having exceeded the Regeneration and Repair ability of surrounding parenchyma cell, bleeds excessive in order to prevent and reduces sense
The risk of dye, body generate it is violent stress protection mechanism generate Coagulation test, immune response and inflammatory reaction, and between promoting
The fibrous connective tissue (extracellular matrix) of matter part repairs tissue of a large amount of hyperplasia to defect, with generation fiber
The pathological change of change.Although although the fibrous connective tissue of hyperplasia has repaired defect, to protect organizer to the greatest extent
The relative fullness of official, but this reparation not only inhibits normal Regeneration and Repair to make this reparation that can not have original organ
Structure and function.Can even be caused due to this excessive, too strong and out of control reparation reaction the fibrosis of organ, hardening from
And function is lost.
In worldwide, the fibrosis of histoorgan is the main reason for many diseases disable, is lethal, organizer
Official's fibrosis plays an important role during the occurrence and development of the related disease of each major organs of human body.According to related system
Meter data shows that in the U.S. patient lethal because of various diseases, histoorgan fibroplasia disease can be attributed to by having close to 45%
Disease.Therefore the progression of fibrosis of control mammal is a variety of one important thinking of disease for the treatment of.
Stress fiber is the microfilament bundle structure being widely present in eukaryocyte, is made of a large amount of microfilaments arranged in parallel, with
Iuntercellular or cell and stromal surface are adhered with substantial connection, occur in cellular morphology, cell differentiation and formation of tissue etc.
Aspect plays a significant role.Its ingredient is actin, myosin, tropomyosin and α actinin.By stress fibre
Dimension, extracellular matrix, cell-membrane receptor (such as integrin), nuclear skeleton and the linker complex of cytoskeleton, nuclear lamina, dyeing
Body skeleton etc. constitutes the main mechanical stress system of cell.Mechanical stress system assigns the regulation of cell mechanical hardness, participates in cell
Induction, transmitting and the generation of inside and outside mechanical force adjust the assembling, distribution and expression of inhereditary material.Mechanical stress systematic steady state
It is one of characteristic index of specific cells.Cell machinery stress system stable state helps to maintain and establish specific cells attribute,
The specific genetic regulation of cell and expression characteristic are maintained and stablized, breaks cell mechanical steady state as a kind of cellular damage and excites
Cellular stress hair is answered, and strong cell injury repair ability is excited.
The present invention is by a kind of without losing without, by destroying cell machinery stress system stable state, exciting in the way of bleeding
Similar to the stress reaction and regenerative response during regeneration, cell can be greatly improved using stress reaction and is lost
Pass repair ability.Since present invention processing is simple, it can be used as raising mammal and mobilize body in body body during lesion and damage
A kind of new method that cell participates in cell turnover, inhibits tissue fibrosis, improves tissue repair and neomorph ability.It can also
New tool as the homologous recombination ability for improving cell background homologous recombination ability and double-strand break mediation.
Summary of the invention
The present invention is the following discovery based on inventor and completes: Myosin inhibitor can destroy cell mechanical stress
Systematic steady state, excitation are greatly mentioned similar to stress reaction and regenerative response during regeneration using stress reaction
High cytogenetics repair ability, and then complete the present invention.
Therefore, in one embodiment, the application the present invention relates to Myosin inhibitor in excitation stress reaction.
In one embodiment, the present invention relates to Myosin inhibitor steady by destroying cell machinery stress system
State excites the application in stress reaction.
In one embodiment, the application the invention further relates to Myosin inhibitor in excitation regenerative response.
In one embodiment, the present invention relates to Myosin inhibitor steady by destroying cell machinery stress system
State excites the application in regenerative response.
In one embodiment, the invention further relates to Myosin inhibitor in improving cytogenetics repair ability
Using.
In one embodiment, the present invention relates to Myosin inhibitor steady by destroying cell machinery stress system
State improves the application in cytogenetics repair ability.
In one embodiment, the excitation, which stress be sent out, should cause to activate high genetic damage reparation hair to be answered, Jin Erti
The up-regulation of high ontology homologous recombination repair gene improves recombination ability.
In one embodiment, the invention further relates to Myosin inhibitor in the Regeneration and Repair for promoting histoorgan
Application.
In one embodiment, the present invention relates to Myosin inhibitor steady by destroying cell machinery stress system
Application in Regeneration and Repair of the state to promote histoorgan.
In one embodiment, the invention further relates to Myosin inhibitor to prepare for exciting stress reaction, excitation
Regenerative response improves cytogenetics repair ability and/or the drug of Regeneration and Repair for promoting histoorgan or answering in reagent
With.
In one embodiment, the invention further relates to Myosin inhibitor destroys cell machinery for passing through in preparation
Stress system stable state come excite stress reaction, excitation regenerative response, improve cytogenetics repair ability and/or promote organizer
The drug of the Regeneration and Repair of official or the application in reagent.
In one embodiment, the invention further relates to Myosin inhibitor is used to treat with stress reaction, again in preparation
Raw reaction, cytogenetics repair ability and/or histoorgan the relevant disease of Regeneration and Repair drug or answering in reagent
With.
In one embodiment, the disease is chorionitis.
In one embodiment, the Myosin inhibitor is (-)-Blebbistatin.
In one embodiment, the organ is liver.
Method of the invention only needs single small molecule or single factor test to handle, easy to operate, reproducible, can be used as excitation
Stress reaction, excitation regenerative response, the one kind for improving cytogenetics repair ability and/or promoting the Regeneration and Repair of histoorgan
New method.
The meaning of term herein is as follows:
DMEM in high glucose: a kind of high glycoform DMEM culture medium (dulbecco's modified eagle medium, DMEM),
The culture medium of i.e. a kind of commercialization containing various glucose and amino acid is developed on the basis of MEM culture medium.
N2B27: one kind is mixed with DMEM/F12 basal medium and neurobasal basal medium with 1:1, is wrapped
The cell culture fluid of the definite ingredients of additive containing N2 and B27 additive, report are conducive to mouse embryo stem cell Godwards classical prescription
To differentiation.
DMEM/F12: a kind of basic culture solution of the commercialization mixed with DMEM culture medium and F12 culture medium 1:1,
Culture suitable for Clonal density.
Neurobasal: be conducive to the commercial basis culture medium of neuronal cell cultures.
GlutaMAX: a kind of cell culture additive can directly substitute the L-Glutamine in cell culture medium.
Dual anti-: penicillin and streptomysin are the common two kinds of antibiotic of cell culture, are prevented thin in cell cultivation process
Bacterium pollution.
A kind of N2 additive: cell culture additive that serum-free is commercialized.
A kind of B27 additive: cell culture additive that serum-free is commercialized.
Detailed description of the invention
Fig. 1 shows that Myosin inhibitor destroys the figure of cell machinery systematic steady state and inducing cell softening.
Fig. 2A shows that bile duct ligation Liver Fibrosis Model control group dmso treatment mouse is dispirited, and mobility subtracts
It is weak, it is more active that processing group mouse is significantly inhibited compared to Myosin inhibitor (-)-Blebbistatin, is full of energy.
Fig. 2 B shows that Myosin inhibitor (-)-Blebbistatin significantly inhibits processing group and has more preferably compared to control group
Fiber macula lutea block.
Fig. 2 C shows that Myosin inhibitor (-)-Blebbistatin significantly inhibits processing group and can improve fibrosis mouse
Survival rate.
Fig. 2 D shows that Myosin inhibitor (-)-Blebbistatin significantly inhibits fibrosis and stockpiles.
Fig. 2 E shows every group of at least 5 mouse statistics degree of fibrosis quantitative results.
Fig. 3 A shows that fibroblastic sample to (-)-Blebbistatin processing people carries out the number of transcript profile sequencing
According to analysis result.
Fig. 3 B and Fig. 3 C, which are shown in 1 day and 2 days these gene expressions mRNA, to be started to lower.It is lured under nerve-inducing system
It leads 22 days, regeneration starting related gene such as FST and nerve to occur related gene obviously raise.
Fig. 4 A shows experiment flow figure.
Fig. 4 B-I shows the effect of experiment of (-)-Blebbistatin for treatment chorionitis.
Fig. 5 shows that experiment flow and (-)-Blebbistatin significantly inhibit the figure of fibrosis.
Fig. 6 shows that cell machinery systematic steady state destroys the figure that the relevant reaction of regeneration induction promotes hepatic tissue Regeneration and Repair.
Fig. 7 A shows the fibroblast of 5-50 micromole (-)-Blebbistatin processing people, 6 hours transcription components
Analysis, DNA replication dna correlation, homologous recombination related gene, mispairing Related to repair gene, Nucleotide Sequence Analysis and base excision are repaired
Multiple genes significantly raises.
Fig. 7 B shows that further analysis finds that nearly all participation homologous recombination correlation important gene all significantly raises.
The adjoint high genetic damage reparation hair of Fig. 8 display excitation stress reaction should improve background homologous recombination gene and repair
The figure of multiple process.
Specific embodiment
Embodiments of the present invention are elaborated and illustrated below by way of specific embodiment, but the following contents should not manage
Solution is to impose any restrictions to the present invention.
One: Myosin inhibitor of embodiment destroys cell machinery systematic steady state and inducing cell softening
Figure 1A is laser confocal scanning schematic diagram, and Figure 1B shows DMSO (control), and (-)-Blebbistatin is (also simple
Referred to as Blebbistatin or Bleb or Ble) (20 micromole) handle the fibroblast of people, and phalloidine changes dyeing, control
Group (lastrow) cell has stress fiber arranged in parallel abundant, and nucleus the smooth of the edge is rounded or oval, experiment
Stress fiber disintegrates after group Ble processing, and nucleus fold occurs in irregular.Fig. 1 C shows Lamin A/C outside DNA
Be with it is stronger be uniformly distributed, have apparent polarity on substrate-the tip of the tongue direction, the lip of the tongue divides Lamin A/C obviously high
In base part.Heterochromatin HP1 albumen shows that control group is significantly stronger than experimental group.Experimental group is greater than Fig. 1 D as the result is shown
Stress effect protein Y AP1/TAZ nuclear localization signal is significantly stronger than cytoplasm and positions in 90% cell, and experimental group drug-treated
Only 35% cell has significantly and positions.Fig. 1 E quantitative PCR measures cell mechanical hardness and regulates and controls relevant albumen small
It is obvious after molecule processing to lower.RNA is transcribed using fluorouracil nucleosides label ribosomes, as Fig. 1 F shows that control group turns
Record active region is focused primarily upon close to nuclear centers, and experimental group then shows that fluorouracil nucleosides label active transcription area exists
It is also distributed close to nucleus periphery, biology repeats three times.Influence of the immunofluorescence dyeing Ble to epigenetic, as a result such as
Fig. 1 G shows that Ble significantly reduces the tri-methylated modification of H3K27 on day 4, further verifies the tri-methylated modification of H3K27
Complex component EZH2 has found the obvious nuclear location of control group EZH2, and Ble processing EZH2 protein level, which is lowered and is shown as cytoplasm, to be determined
Position, as shown in fig. 1H.
Embodiment two: cell machinery systematic steady state destroys the survival rate for improving fibrosis mouse
Experiment using the purchase of ICR mouse in Si Beifu (Beijing) Bioisystech Co., Ltd, elbow tweezers, finger tweezers,
Scissors, needle holder, sewing needle, suture are purchased from Ya Suwang commerce and trade Co., Ltd, and antibiotic is purchased from Gibco (15240-062).
Select 8 week old female mices, 24 hours fasting for solids and liquids before experiment, using 5% chloral hydrate anesthesia mouse, 8 milliliters/thousand
Gram intraperitoneal injection, mouse is fixed after anesthesia, notch in abdomen exposes internal organs, finds the duodenum for getting close to stomach end, gently leads
It draws duodenum to find out bile duct, carefully separates bile duct with tweezers, ligature bile duct with suture, antibiotic is added dropwise, rear suture is closed
Close abdominal cavity.12 hours fasting for solids and liquids after operation.
Postoperative 14-20 days, it can be observed that mouse skin is in yellow green, it is administered treatment.It is seeped using implanted capsule
Saturating press pump, is sustained two weeks (ALZEN, 1002), drug concentration by 0.25 micro- l/h.Using 5% chloral hydrate anesthesia mouse, 8
Ml/kg intraperitoneal injection, fixes mouse, notch in abdomen exposes internal organs, carefully pushes liver and intestines open, will permeate after anesthesia
Press pump is implanted into abdominal cavity, and antibiotic is added dropwise, and abdominal cavity is closed in rear suture.
Result figure 2A shows that bile duct ligation Liver Fibrosis Model control group dmso treatment mouse is dispirited, movable energy
Power weakens, and it is more active to significantly inhibit processing group mouse compared to Myosin inhibitor (-)-Blebbistatin, full of essence
Power.Fig. 2 B shows that Myosin inhibitor (-)-Blebbistatin significantly inhibits processing group and has better fibre compared to control group
Macula lutea block is tieed up, while Myosin inhibitor (-)-Blebbistatin significantly inhibits processing group can improve depositing for fibrosis mouse
Motility rate, as shown in Figure 2 C, the result as a result from 5 mouse.Sirius red stains Myosin inhibitor (-)-as the result is shown
Blebbistatin significantly inhibit but tubal ligation caused by fiber stockpile, as shown in Fig. 2 D-E.
Embodiment three: cell machinery systematic steady state destroys relevant stress send out of regeneration induction and answers and regenerate starting and tissue
Correlated response occurs
(-)-Blebbistatin handles the fibroblast of people, collects processing 0 hour, 6 hours, 1 day, 2 days samples respectively
Product carry out transcript profile sequencing, and data analysis as shown in Figure 3A, is inducing 6 hours a large amount of HSP70 families and HSP90 etc. one of early stage
As the relevant gene upregulation of response.Start to lower 1 day and 2 days these gene expressions mRNA.Under nerve-inducing system
Induction 22 days, regeneration starting related gene such as FST and nerve to occur related gene obviously raise, as shown in Fig. 3 B and Fig. 3 C.
Example IV: cell machinery systematic steady state destroys application of the relevant reaction of regeneration induction in treatment chorionitis
Chorionitis is that a kind of hardened in turn with limitation or the fibrosis of diffusivity skin and internal organs with atrophy is special
The connective tissue disease of sign.This disease can cause Multisystem damage, its definite cause of disease and pathogenesis are still not known at present, also
Without effective treatment means.The present invention using bleomycin (bleomycin, BLM) locally injecting in ICR mouse back at
Function induces mouse skin hardening, and intraperitoneal administration Myosin inhibitor (-)-Blebbistatin is treated, and experiment flow is as schemed
Shown in 4A.
Bleomycin inject 1 week after, back of mice injection site skin i.e. occur pachyderma be hardened, poor flexibility, hair
It has no the change of growth, terminates up to injecting, while scleroma, incrustation occur in injection site, it is adjoint superficial ulcer occur.It is raw
Manage the injection zone hair continued growth of saline control group back of mice shaving, the performance for having no obvious sclerosis of the skin, thickening,
As shown in Figure 4 B.Compared to control group intraperitoneal administration DMSO, intraperitoneal administration Myosin inhibitor (-)-Blebbistatin (1-
3mg/kg/ days, it is dissolved in 2%DMSO+25%PEG400+2%Tween 80+ddH2O scleroma, incrustation journey can) be substantially reduced within 3 weeks
Degree, with the superficial ulcer of smaller extent, as shown in Figure 4 B.Further experimental verification Myosin inhibitor (-)-
Blebbistatin promotes scleroma area's hair growth such as Fig. 4 C.Significant decrease epidermis and dermis thickness, promotion hair follicle and body of gland
Number, such as Fig. 4 D-F.(-)-Blebbistatin significantly reduces stockpiling for interstitial fibers as the result is shown for stock-dye, such as Fig. 4 G
Arrow instruction.Ki-67 coloration result shows that (-)-Blebbistatin promotes hair follicle cell proliferation, so that hair follicle be promoted to be formed
Such as Fig. 4 H.Make us feeling surprised, (-)-Blebbistatin can promote area's nerve destiny appearance of hardening, some cell tables
Up to Marker NeuN such as Fig. 4 I of the mature neuron of nuclear location, implies whether the induction of neural destiny has and help damage
Wound is repaired and regeneration is up for further studying.
Embodiment five: cell machinery systematic steady state destroys the relevant reaction of regeneration induction and is promoting hepatic tissue Regeneration and Repair
Using
CCl is established in induction4Liver injury model
Experiment is bought using ICR mouse in Si Beifu (Beijing) Bioisystech Co., Ltd, and carbon tetrachloride is purchased from Aladdin
Reagent (Shanghai) Co., Ltd. (C131583-1L), corn oil are purchased from Sigma (C8267).Experiment flow is as shown in Figure 5A, choosing
With 8 week old female mices, is raised one week in barrier, be randomly divided into two groups, inject CCl respectively4/ corn oil (dose volume 2:5) and
Corn oil, 2 ml/kg dosage, twice a week, continuous 8 weeks.5% chloral hydrate anesthesia mouse is used after 8 weeks, takes out part
Liver page, is fixed with 4% paraformaldehyde, is dehydrated waxdip, is carried out using Picro-Sirius red (Y-Y-R20384-100ML) kit fine
Dimensionization dyeing, wherein collagenous fibres take on a red color, as shown in Fig. 5 B, C.After identifying modeling success, it is administered treatment.Using abdomen
The mode of chamber injection and tail vein injection carries out.Small molecule is dissolved in dimethyl sulfoxide, and with normal saline dilution, small molecule is dense
1 milligrams per kilogram of degree daily, or sequentially adds that (> 1mg/kg sequentially adds 5%DMSO+30% propylene glycol+3%Tween
80+ddH2O or 2%DMSO+25%PEG400+2%Tween 80+ddH2O), 1 milliliter of syringe draws drug, with subcutaneous note
Emitter punctures skin and stomach wall flesh, and liquid injection is careful not to hurt diaphragm and other organs, stops one to intraperitoneal
Needle can be pulled out again, and fluid seepage is avoided to come out.Mouse is fixed in fixator when tail vein injection, tail is straightened and is tightened, is infused
Stopped blooding after penetrating with cotton.Continuous injection 7 days, stops injection.Sirius red stains identification is carried out again, as a result such as Fig. 5 D, E institute
Show, Myosin inhibitor (-)-Blebbistatin significantly inhibits fibrosis.
The identification of liver function Regeneration and Repair
Further hybridize mouse using Alb-cre × mTmG, mature hepatocytes express GFP, and CCl4 carries out fibrosis and makes
Mould, after drug therapy processing, most of cell for expressing GFP expresses Alb, and the cell of mono- a part expression GFP of DMSO not table
Up to Alb, illustrate that Myosin inhibitor (-)-Blebbistatin can prevent liver function in course of liver damage from losing, such as Fig. 6 A institute
Show.Blood biochemistry analysis (empty stomach 12-16 hours before sampling) further confirms that Myosin inhibitor (-)-Blebbistatin can drop
The content of low hepatic injury index such as ALT, AST and GGT etc., as shown in Figure 6B.Further dyeing identification discovery (-)-
Blebbistatin remarkably promotes cell Proliferation after hepatic injury, and as presented in figs. 6 C-D, and the cell reduced in course of liver damage withers
It dies, as shown in Fig. 6 E-F.In conclusion Myosin inhibitor (-)-Blebbistatin is by inhibiting hepatic injury fibrosis, promoting
Into hepatocyte growth and Apoptosis promotion liver regeneration is reduced, to maintain liver function.
Embodiment six: (-)-Blebbistatin excitation stress send out the high genetic damage reparation hair that stress be lived and answer, and improve
The gene upregulation of ontology homologous recombination repair
As shown in Figure 7 A, 5-50 micromole (-)-Blebbistatin handles the fibroblast of people, 6 hours transcript profiles
Analysis, DNA replication dna correlation, homologous recombination related gene, mispairing Related to repair gene, Nucleotide Sequence Analysis and base excision
Revision points significantly raise.As shown in Figure 7 B, further analysis finds nearly all participation homologous recombination correlation important gene all
Significant up-regulation.
Embodiment seven: the high genetic damage reparation hair of (-)-Blebbistatin excitation stress reaction activation is answered, and is being mentioned
Application during the gene of high ontology homologous recombination repair
Experiment flow is as shown in Figure 8 A, the reporter plasmid system of homologous recombination repair is constructed first, only when external source is homologous
Cell fluoresced green when homologous recombination occurs for sequence and cell-isogenic segment.It is transferred to mRuby red fluorescent protein simultaneously, is used
Come mark transfection cell efficiency, judged by the ratio of flow cytometer showed green fluorescent protein and mRuby occur homologous recombination
Efficiency.Control group after liquid is changed in transfection 18h with DMSO handle 4h, experimental group respectively after liquid is changed in transfection 18h, for 24 hours, 36h use
(-)-Blebbistatin handles 4h, 72h flow cytometer showed HDR efficiency after transfection.(-)-Blebbistatin is significant as the result is shown
Background homologous recombination efficiency is improved, as shown in 8B-C.It is that oxidation is answered since Nrf2 is that response to oxidative stress significantly raises gene
Sharp effect protein, homologous recombination can be improved by further verifying stress reaction by small molecule Nrf activator simulation oxidative stress
Ability, as shown in Fig. 8 D-E.Experiment flow is as follows, and control group is handled after liquid is changed in transfection with DMSO, and experimental group changes liquid in transfection
It is handled afterwards with 250nM RTA408,72h flow cytometer showed HDR efficiency after transfection.
Foregoing merely illustrate the principle of the present invention, it should be appreciated that the scope of the present invention is not intended to as described herein
Illustrative aspect, and should include the equivalent of all currently known and following exploitation.Further, it is noted that not departing from this
Under the premise of inventive technique principle, it can also make several improvements and modify, these are improved and modification should also be considered as the present invention
Range.
Claims (12)
1.Myosin inhibitor is in excitation stress reaction, excitation regenerative response and/or improves answering in cytogenetics repair ability
With.
2.Myosin inhibitor is in preparation for exciting stress reaction, excitation regenerative response and/or improving cytogenetics reparation energy
The drug of power or the application in reagent.
3. such as application of any of claims 1-2, which is characterized in that the excitation stress reaction, excitation regeneration are anti-
Answering and/or improve cytogenetics repair ability is realized by destroying cell machinery stress system stable state.
4. application as claimed in any one of claims 1-3, which is characterized in that the excitation stress send out should cause to activate it is high
Genetic damage reparation hair is answered, and then improves the up-regulation of ontology homologous recombination repair gene, improves recombination ability.
Application of the 5.Myosin inhibitor in the Regeneration and Repair for promoting histoorgan.
6.Myosin inhibitor is preparing the application in drug or reagent for promoting the Regeneration and Repair of histoorgan.
7. the application as described in any one of claim 5-6, which is characterized in that it is described promote histoorgan Regeneration and Repair be
It is realized by destroying cell machinery stress system stable state.
8.Myosin inhibitor is related to stress reaction, regenerative response and/or cytogenetics repair ability for treating in preparation
Disease drug or the application in reagent.
9.Myosin inhibitor is in the drug or reagent that preparation is used to treat disease relevant to the Regeneration and Repair of histoorgan
Application.
10. the application as described in any one of claim 8-9, which is characterized in that the disease is chorionitis.
11. such as application of any of claims 1-10, which is characterized in that the Myosin inhibitor is (-)-
Blebbistatin。
12. the application as described in any one of claim 5-7,9-11, which is characterized in that the organ is liver.
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| CN201810082885.2A CN110090300A (en) | 2018-01-29 | 2018-01-29 | A kind of method and its application for the Regeneration and Repair promoting histoorgan |
| EP19743774.2A EP3747440A4 (en) | 2018-01-29 | 2019-01-29 | Method for destroying cellular mechanical homeostasis and promoting regeneration and repair of tissues and organs, and use thereof |
| JP2020562820A JP2021512173A (en) | 2018-01-29 | 2019-01-29 | Methods and uses that disrupt the mechanical homeostasis of cells and promote the regeneration and repair of tissue organs |
| CN201980005648.9A CN111405898B (en) | 2018-01-29 | 2019-01-29 | Method for destroying cell mechanical homeostasis and promoting tissue and organ regeneration and repair and application thereof |
| PCT/CN2019/073622 WO2019144967A1 (en) | 2018-01-29 | 2019-01-29 | Method for destroying cellular mechanical homeostasis and promoting regeneration and repair of tissues and organs, and use thereof |
| US16/965,401 US11622964B2 (en) | 2018-01-29 | 2019-01-29 | Method for destroying cellular mechanical homeostasis and promoting regeneration and repair of tissues and organs, and use thereof |
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| CN110090301A (en) * | 2018-01-29 | 2019-08-06 | 中国科学院动物研究所 | A kind of method and its application for destroying cell mechanical steady state |
| US11622964B2 (en) | 2018-01-29 | 2023-04-11 | Institute Of Zoology, Chinese Academy Of Sciences | Method for destroying cellular mechanical homeostasis and promoting regeneration and repair of tissues and organs, and use thereof |
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Cited By (2)
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| CN110090301A (en) * | 2018-01-29 | 2019-08-06 | 中国科学院动物研究所 | A kind of method and its application for destroying cell mechanical steady state |
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