CN108159051A - Application of the 3-MA in the drug for preparing treatment Subretinal Fibrosis - Google Patents
Application of the 3-MA in the drug for preparing treatment Subretinal Fibrosis Download PDFInfo
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- 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
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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Abstract
The present invention relates to application of 3 methyl adenines in the drug for preparing treatment Subretinal Fibrosis (SRF).Subretinal Fibrosis animal model is initially set up, verifies being successfully established for the model；Key effect of the macrophage in SRF is proved secondly by systemic injection macrophage scavenger chlorine disodium hydrogen phosphate liposome；Macrophage is navigated to as a result, autophagy is activated then in conjunction with finding that eyeball tissue autophagy activity is raised early period, reapplies 3 MA intraperitoneal injections, influences of 3 MA of observation analysis to macrophage autophagy activity, quantity, the parting that polarizes；Finally explore the associated signal paths that 3 MA play a role to macrophage and fibrosis.Experimental result shows that 3 MA intraperitoneal injections can significantly inhibit Subretinal Fibrosis, it is by inhibiting macrophage autophagy active, macrophage quantity is reduced, adjusts macrophage subsets, the activation that macrophage promotees fibrosis signal path is inhibited to play anti-fibrosis effect.
Description
Technical field
The present invention relates to biomedicine technical field, specifically, being that 3-MA is treated in preparation under retina
Application in the drug of fibrosis.
Background technology
Age-related macular degeneration (age-related macular degeneration, AMD), also known as age-related macular
One of the main reason for denaturation is 50 years old or more adult's blinding.In recent years, AMD incidence is in the gesture increased year by year.Developing
Middle country, the incidence of AMD is about 7%, in developed country, it is contemplated that will increase by 50% to the year two thousand twenty AMD patient, and suffer from close to 23%
Person's eyesight completely loses.It is clinically dryness (dry AMD) and moist (wet AMD, wAMD) by AMD point, the former accounts for AMD patient
80%-90%, the latter accounts for 10%-15%.Although moist AMD is accounted for, patient's toatl proportion is less, and about 90% AMD is relevant to be regarded
Power is lost related with moist AMD.The Visual outcome of wAMD patient is poor, with choroidal neovascularization (choroidal
Neovascularization, CNV) and subsequent Subretinal Fibrosis (subretinal fibrosis, SRF) formation,
Progression of the disease is rapid.Although common anti-vascular endothelial growth factor (anti-vascular endothelial growth
Factor, anti-VEGF) drug therapy can improve the eyesight of wAMD patient, but research shows that about half of in 2 years
The eyes of anti-vegf treatment can form retina fibrous scar.SRF leads to retinal light injury photoreceptor, retinal color
Plain epithelium (retinal pigment epithelium, RPE) and the irreversible damage of choroidal artery, so as to cause eyesight
Permanent loss, therefore there is an urgent need to strengthen the research to SRF, research and development are a kind of effectively to be prevented and intervene the drug of SRF to protect
Hinder anti-vegf curative effect, improve wAMD patient's vision prognosis.
At present, clinically there is no the means for for SRF effectively prevent and intervene.Basic research shows that SRF may
It is due to caused by excessive wound healing reaction, and lasting damage or inflammatory conditions may be to cause excessive damage healing anti-
The premise answered.Studies have found that visible a large amount of macrophages infiltrations in tunica fibrosa under the retina of AMD patient's corpse eye, and
Macrophages infiltration is had also discovered in animal model in CNV regions, illustrates that immunization inflammatory reaction that macrophage participates in may be with
The formation of Subretinal Fibrosis is related.The fibrosis research of liver, lung tissue in recent years be also directed to the morbidity of fibrosis with
Participation, Epithelial and stromal conversion, oxidative stress and the associated signal paths activation of inflammatory reaction etc. are related.Therefore, from the morbidity of SRF
Mechanism is set out, and it is critical issue urgently to be resolved hurrily to find the crucial target spot of anti-fibrosis generation and carry out effective prevention.
Autophagy is the highly conserved process that degradation recycling is carried out to intracellular matter during eucaryote is evolved, a variety of
It plays a significant role in physiology and pathologic process, numerous studies show that autophagy participates in the occurrence and development of fibrosis in recent years.This
Invention carries out that common autophagy inhibitor 3-MA (3-Methyladenine, 3-MA) is injected intraperitoneally in animal body,
It was found that 3-MA significantly inhibits Subretinal Fibrosis.We further study 3-MA by vitro and in vivo experiments and inhibit
The mechanism of SRF.
Invention content
First purpose of the present invention is for deficiency of the prior art, provides 3-MA and is regarded in preparation treatment
Application under nethike embrane in the drug of fiber.
Second object of the present invention is for deficiency of the prior art, provides 3-MA and is preparing treatment always
Application in the drug of year macular degeneration.
Third object of the present invention is for deficiency of the prior art, provides 3-MA in reagent preparation
Application.
Fourth object of the present invention is for deficiency of the prior art, provides a kind of medicine of anti-Subretinal Fibrosis
Compositions.
To realize above-mentioned first purpose, the technical solution adopted by the present invention is that：
The application of 3-MA in medicine preparation, the drug are used to treat Subretinal Fibrosis.
As the preferred embodiment of the present invention, the Subretinal Fibrosis refers to that age-related macular degeneration is relevant
Subretinal Fibrosis.
As the preferred embodiment of the present invention, the formation of the Drug inhibition Subretinal Fibrosis.
As the preferred embodiment of the present invention, the drug is reduced huge by inhibiting macrophage autophagy active
Phagocyte quantity adjusts macrophage subsets, and the activation that macrophage promotees fibrosis signal path is inhibited to play anti-fibrosis work
With.
As the preferred embodiment of the present invention, the administering mode of the drug is drug administration by injection.
To realize above-mentioned second purpose, the technical solution adopted by the present invention is that：
Application of the 3-MA in the drug for preparing treatment age-related macular degeneration, under the Drug inhibition retina
The formation of fibrosis.
As the preferred embodiment of the present invention, the age-related macular degeneration refers to moist AMD.
To realize above-mentioned third purpose, the technical solution adopted by the present invention is that：
Application of the 3-MA in reagent preparation, the reagent are used to adjust macrophage phenotype.
To realize above-mentioned 4th purpose, the technical solution adopted by the present invention is that：
A kind of pharmaceutical composition of anti-Subretinal Fibrosis, described pharmaceutical composition include 3-MA and
Pharmaceutically acceptable carrier or auxiliary material, the Subretinal Fibrosis refer to fiber under the relevant retina of age-related macular degeneration
Change, described pharmaceutical composition inhibits the formation of Subretinal Fibrosis.
The present invention initially sets up Subretinal Fibrosis animal model, and that verifies the model is successfully established (Fig. 1), secondly logical
Crossing systemic injection macrophage scavenger chlorine disodium hydrogen phosphate liposome proves key effect (Fig. 2) of the macrophage in SRF, so
It combines afterwards and finds that eyeball tissue autophagy activity is raised early period as a result, autophagy activation is navigated to macrophage (Fig. 3) first, then
It is injected intraperitoneally using 3-MA, observation analysis 3-MA is to macrophage autophagy active (Fig. 3), quantity (Fig. 4), polarization parting (Fig. 5)
Influence, finally explore the associated signal paths (Fig. 6) that play a role to macrophage and fibrosis of 3-MA.
The invention has the advantages that：
1st, present invention firstly discovers that 3-MA intraperitoneal injections can significantly inhibit Subretinal Fibrosis.
2nd, 3-MA reduces macrophage quantity, adjusts macrophage subsets, inhibit by inhibiting macrophage autophagy active
The activation that macrophage promotees fibrosis signal path plays anti-fibrosis effect.
3rd, find whole body application 3-MA without obvious toxic-side effects.
Description of the drawings
Attached drawing 1 is the time trend of damage from laser MODEL C NV and SRF as a result, 35 days choroids are new after damage from laser
Angiogenic subsides and Subretinal Fibrosis generation is notable.
Attached drawing 2 removes experimental result for macrophage, and macrophage removes the formation for reducing SRF.
The influence to macrophage activity is injected intraperitoneally for 3-MA for attached drawing 3, and display 3-MA can significantly inhibit macrophage certainly
Bite activity.
The influence to macrophage quantity is injected intraperitoneally for 3-MA for attached drawing 4, and display 3-MA considerably reduces SRF and surrounding is huge
Phagocyte quantity does not influence Macrophage in Spleen quantity but.
The influence for the parting that polarizes to macrophage is injected intraperitoneally for 3-MA for attached drawing 5, shows around SRF with M2 type macrophages
Based on recruitment, 3-MA regulation and control macrophage polarization partings.
Attached drawing 6 is the associated signal paths that 3-MA plays a role to macrophage and fibrosis, and display 3-MA passes through inhibition
Macrophage STAT3/TGF- β2Signal path inhibits Subretinal Fibrosis.
Specific embodiment
The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair
It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after the content of the invention recorded has been read, art technology
Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Fixed range.
Pharmaceutical composition
Method according to the present invention, 3-MA can prepare the pharmaceutical composition that anti-Subretinal Fibrosis is formed
Object, the pharmaceutical composition can be made of active constituents of medicine 3-MA and pharmaceutically acceptable carrier.
The method that medical domain routine can be used in the pharmaceutical composition, using 3-MA as active constituent,
Various dosage forms are made with pharmaceutically acceptable auxiliary material.When for taking orally, conventional solid pharmaceutical preparation such as piece can be prepared into
Agent, pulvis or capsule etc.；During for injecting, injection can be prepared into.In various preparations, the weight of active constituent contains
Measure is 0.01%~99.9%.
The pharmaceutical composition can be used for treating the disease of Subretinal Fibrosis.Such as mouth can be passed through by dosage form
The approach such as clothes, intraperitoneal injection, hypodermic injection, intravenous injection, intramuscular injection, mucous membrane medication are applied to the individual for the treatment of, institute
It can be human or animal to state individual.
In the present invention, " pharmaceutically acceptable " herein refers to a kind of substance, such as carrier or dilution, will not make 3- methyl
The bioactivity or property of adenine disappear, and relative nontoxic.Such as something is added in, undesired biotic influence will not be caused
Or it interacts in harmful manner with 3-MA.
In the present invention, the treatment, symptom or situation including mitigating, inhibiting or improving disease；Inhibit disease or symptom
Generation, such as control the development of disease or situation；Mitigate disease or symptom；Disease or symptom is made to decline.It is described to control in this hair
It treats, espespecially inhibits the formation of Subretinal Fibrosis.
Embodiment
In the present embodiment, the fibrosis model in the case where eyeball of mouse Posterior pole implementation 532nm laser establishes Mouse Retina is visited
Beg for therapeutic effects of the purine compound 3-MA to Subretinal Fibrosis.Specific experiment process is as described below.
1.1 experimental animal
6-8 week old C57BL/6J healthy male mices 50,100, weight is 20~25g.
1.2 reagents and instrument
3-MA compound, chlorine disodium hydrogen phosphate liposome, Sodium fluorescein, primary antibody (F4/80, IB4-
Lectin, Fibronectin, LC3, Atg5, Becline-1, YM-1, Arg-1, transforming growth factor TGF-β2, signal transduction
With activating transcription factor STAT3 etc.), fluorescence secondary antibody etc..Coherent laser therapeutic apparantus, Heidelberg fluoroscopic visualization fundus imaging
Instrument, Olympus fluorescence microscopes, Leica Laser Scanning Confocal Microscopes, freezing microtome etc..
1.3 experimental method
1.3.1 the foundation of Subretinal Fibrosis model
In order to induce SRF, we carry out Fundus laser damage to male C 57 BL/6 J mouse using 532nm laser and observe
By 35 days.In brief, we have carried out laser photocoagulation with transparent cover slides effect contact lense at the 0th day to mouse eyeground
(wavelength 532nm, energy 120mW, interval time 100ms, 50 μm of spot diameter)；At the 7th day, the 21st day and the 35th day respectively into
Row observation and materials.The Mouse Retina and tela chorioidea for receiving the damage of 8-10 laser point around the optic nerve of eyeground are used for
Western Blots experimental analyses, the Mouse Retina and choroid of 4 laser points damages of receiving are used for Immunofluorescence test.
1.3.2 macrophage removes experiment
In order to remove macrophage, the different time points clodronate liposome Clodrosome after laser photocoagulation
(SKU：8901, Encapsula Nanosciences, 50mgkg-1) it is injected intraperitoneally.The mouse that terminal is 7 days is observed,
Receive liposome injection within 1,3 and 6 days after laser photocoagulation；The mouse that terminal is 35 days is observed, it is every after being injected three times before receiving
Week receives a shot until observing 35 days again.Control group injects the liposome Encapsome for not wrapping up chlorine disodium hydrogen phosphate.
1.3.3 3-MA is injected intraperitoneally
3-MA powder (M9281, Sigma-Aldrich, Saint Louis, USA) is dissolved in 65 DEG C of deionized waters, concentration
For 30mgmL-1, dilute 10 times with 0.9% brine and carry out mouse peritoneal injection, dosage 15mgkg-1, frequency is every two days
Injection is primary.Isometric 0.9% physiological saline is injected intraperitoneally in control group.
1.3.4 fluorescence blood light radiography
7 days after modeling, fluorescein angiography (FA) is carried out within 21 days, 35 days, to observe the severity of CNV leakages.
Every mouse peritoneal injects 10% fluorescein sodium (fluorescein) 0.05mL；Use digital fundus photography machine (Heidelberg retinal blood
Pipe radiography, Vista, CA) Fundus angiography photo is captured, and calculate the average signal strength of each CNV lesions.By view
The signal strength of film blood vessel is defined as 1, and avascular area domain signal strength is defined as 0, using Image softwares to being oozed at damage from laser
Leakage signal slightly measures, signal strength 0 (most dark) to the digital representation of 1 (most bright) of each pixel.
1.3.5 choroid tile
Respectively the 7th day after damage from laser, the 21st day, the 35th day, cardiac perfusion is carried out to mouse with 4% paraformaldehyde.
2 hours are fixed after winning eyeball of mouse, is placed in PBS.Anterior ocular segment and retinal tissue are removed under microscope, by remaining cup-shaped
RPE train of thought film composites are paved after cutting 4-6 radial incision.
1.3.6 immunofluorescence dyeing and quantization
Tile to be contaminated or slice confining liquid (lowlenthal serum of 0.3%Triton+5%) are closed one hour, Ran Houyong
Confining liquid prepares primary antibody, and 4 spend night.Next day discards primary antibody, and PBS is cleaned three times, and fluorescence secondary antibody, incubation at room temperature are prepared with confining liquid
1 hour, secondary antibody is discarded, PBS carries out DAPI core dyes after cleaning one time, and mounting is simultaneously seen under fluorescence microscope or Laser Scanning Confocal Microscope
It examines and takes pictures.In order to quantify macrophage, we mark nucleus using DAPI (blue), and macrophage is marked using F4/80 (red)
Cell.It is considered as just macrophage that red and blue cell is only marked simultaneously.For quantitative and target macrophages
In autophagy activity, we using F4/80 (red) mark macrophage, use DAPI (blue) label nucleus, use
LC3B (green) shows autophagy activation, and the dotted particles of LC3B (green) are considered as the LC3 forms of activation.In order to quantify CNV and
SRF areas, we mark CNV using IB4, and SRF, the survey carried using fluorescence microscope system are marked using Fibronectin
Amount software measures.Every photo at least measures three times, then for statistical analysis with average value.
1.3.7 immunoblot experiment
The 7th day after modeling, the 21st day, the 35th day excessive anaesthesia execution mouse 5, bilateral eyeball, microscope are extractd
It is lower to cut off cornea along corneoscleral junction, crystalline lens and vitreum are removed, detaches retina choroid tissues, view is extracted with lysate
Membrane tissue total protein carries out albuminous degeneration, then electrophoresis, transferring film, adds corresponding 4 DEG C of overnight incubations of primary antibody.Next day, after TBST is washed
It is incubated 1 hour with corresponding secondary antibody room temperature, TBST is imaged after cleaning three times using electrochemical luminescence kit.
1.3.8 cell culture
Mouse macrophage RAW 264.7 is purchased from ATCC (Manassas, VA, USA), and is containing 10%FBS and 1%
It is cultivated in the DMEM of dual anti-(penicillin, streptomysin).In order to detect whether 3-MA influences protein expression and macrophage polarization,
RAW cells are divided into IL-4 (404-ML, 100ng/ml, RD) individually processing group, IL-4 joint 3-MA (5mM) processing groups, processing
Then it collects cell within 48 hours and is used for Western blotting.In order to detect the effect of STAT3 signal paths, by RAW cells JAK2
Tyrosine kinase inhibitor AG490 (S1143,30 μM, Selleck) STAT3 to be blocked to activate, then collection is thin within 24 hours for processing
Born of the same parents carry out Western Blots experiments.
1.4 statistical procedures
Data statistic analysis is carried out using SPSS20.0 statistics softwares, measurement data is examined through Shapiro-Wilk in just
State is distributed, and is represented with mean ± standard deviation；Then it is examined through Levene and checks homogeneity of variance.To CNV bled signals intensity, CNV
And the correlated results of SRF areas, macrophage quantity etc., when meeting homogeneity of variance, each group is compared using one-way analysis of variance
Whole difference；During heterogeneity of variance, examined using Dunnet t.P<0.05 to be considered difference statistically significant.
The time trend of 2.1 damage from laser MODEL C NV and SRF
The results are shown in Figure 1.Our data show that the 7th day after damage from laser, Fluorescein Leakage signal strength reaches peak
It is worth (0.91 ± 0.05), then started to weaken (0.42 ± 0.10) at the 21st day, be only capable of seeing within the 35th day after laser and ooze on a small quantity
Leakage signal (0.23 ± 0.05) (Figure 1A, 1B), therewith it is consistent be CNV in choroid tile area change：The 7th after modeling
Its CNV areas maximum (0.034 ± 0.003mm2), (0.021 ± 0.002mm is obviously reduced in CNV areas within the 21st day2), by the 35th day
It significantly reduces but still there is part to retain (0.011 ± 0.001mm2), and SRF areas after modeling the 7th day to the 35th day in gradual
Increase trend (Figure 1A, 1C).The phenomenon that this CNV gradually subsides and gradually increased with SRF and the CNV late periods of nAMD patient
Natural history is very similar.The above results illustrate damage from laser mouse eyeground model can fibrosis under induced retinal well,
The 35th day time point that pathomechanism can occur as research Subretinal Fibrosis after damage from laser.
2.2 macrophages remove experimental result
The results are shown in Figure 2.Mouse spleen immunofluorescence results show relative to Encapsome groups, Clodrosome tools
Have an apparent effect for removing whole body macrophage, the 7th day clearance rate up to more than 90%, the 35th day clearance rate up to 80% with
Upper (Fig. 2A).The 7th day after modeling, relative to Encapsome groups, the number of macrophages around Clodrosome group fibrosed tissues
Amount significantly reduces (be respectively 40.8 ± 6.3,4.2 ± 3.4), CNV areas be also obviously reduced (be respectively 0.037 ±
0.0085mm2,0.003 ± 0.011mm2), difference has statistical significance (P<0.05) (Fig. 2 B, 2D).The 35th day after modeling,
Chlorine disodium hydrogen phosphate liposome group damage around macrophage quantity be considerably less than control group (be respectively 34.4 ± 4.1,3.4
± 3.0), SRF areas significantly reduce (being respectively 0.045 ± 0.0095mm2,0.004 ± 0.010mm2), and difference has statistics
Learn meaning (P<0.05) (Fig. 2 B, 2E).The above results prompt macrophage to have important work in Subretinal Fibrosis is formed
With.
2.3 3-MA can significantly inhibit macrophage autophagy
The results are shown in Figure 3.Western Blots the result shows that, behind damage from laser mouse eyeground, as time went on,
Autophagy activity gradually rises (Fig. 3 A), and immunofluorescence results further demonstrate the macrophage that autophagy activity is positioned at around damage
In cell (Fig. 3 B).This illustrates that the macrophage around damage is in autophagy state of activation.And then by the way that 3-MA is injected intraperitoneally,
Autophagy GAP-associated protein GAP such as LC3, Atg5, Becline-1 expression is substantially reduced, and LC3 in macrophage relative to control group
The content of particle also significantly reduces (being respectively 37.3 ± 4.3,3.0 ± 2.1) (P<0.05) (Fig. 3 C).The above results are prompted
3-MA intraperitoneal injections can significantly inhibit macrophage autophagy activity.
2.4 3-MA can obviously reduce damage peripheral macrophage quantity
The results are shown in Figure 4.Immunofluorescence results are shown, the 35th day after modeling, around 3-MA processing group fibrotic scars
The quantity of F4/80 positive cells significantly reduces (being respectively 41.7 ± 9.8,8.8 ± 5.5), while fibrosis area is also bright
It is aobvious to reduce (being respectively 0.048 ± 0.016mm2,0.003 ± 0.001mm2), the statistically significant (P of difference<0.05) (figure
4A, 4B, 4C).In addition, in order to inquire into the molecular mechanism of 3-MA effects, we determine GAP-associated protein GAP with Western Blots
Expression.First, we demonstrate that tables of the 3-MA to CNV GAP-associated protein GAPs VEGFA and fibrosis GAP-associated protein GAP CollagenI albumen
What is reached is inhibited.Secondly, we, which have detected, promotees fibrosis factor TGF-β2Protein expression situation, find 3-MA can press down
Raised TGF-β during fibrosis processed2Horizontal (Fig. 4 D).Meanwhile we have detected 3-MA processing to macrophage in spleen
Quantity and the influence of distribution do not find there is significant difference (4E) with Normal group, illustrate that 3-MA whole bodies are applied to normal
Organ has no obvious toxic-side effects.These, can be by inhibiting correlation factor the result shows that long term systemic application 3-MA is comparatively safe
Expression inhibiting macrophage recruitment, so as to effectively inhibit CNV and fibrosis.
The controllable macrophage subsets of 2.5 3-MA
In order to determine to be gathered in the hypotype of fibrosis region peripheral macrophage, we carry out train of thought in the 35th day after laser
Film tile marks M2 type macrophage marker protein Y M-1, macrophage is marked using F4/80, the experimental results showed that fibrosis
Most of macrophages are YM-1 positive (Fig. 5 A) around tissue.In addition, in order to study whether 3-MA influences macrophage Asia
Type, we carry out immunofluorescence in the 35th day after laser to retinal slice.YM- can be observed at damage from laser position in control group
1 signal, and be mainly distributed in F4/80 positive cells, and 3-MA processing group YM-1 signals significantly reduce (Fig. 5 B).In order into one
Step confirms influences of the 3-MA to macrophage subsets, we have detected in vivo and M2 macrophage specific genes in vitro
The expression of (YM-1 and Arg-1).Compared with the control group, two M2 specific genes especially YM-1,35 the skys after laser
It adjusts, and significantly (Fig. 5 C) is lowered after 3-MA processing.In RAW cells, we handle 24 using IL-4 induction M2 polarization in IL-4
Hour after YM-1 and Arg-1 expression increase, but 3-MA handle 24 hours after its expression be substantially reduced (Fig. 5 D).The above result shows that
3-MA can be by inhibiting M2 Phenotypic Changes to inhibit the rush Fibrosis parameters of M2 type macrophages.
2.6 3-MA, which can inhibit, promotees the activation of fibrosis signal path
Research shows that in wAMD patient in circulating monocytic cell there are the phenomenon that STAT3 activation, therefore we have detected
The level of STAT3 signal paths in macrophage.We have found that control group pSTAT3 (p-STAT3) expression is in fibre
It is apparent during dimensionization to increase, and p-STAT3 expresses apparent lower after 3-MA processing.Total STAT3 protein expression levels are at two groups
Between without significant change (Fig. 6 A).In addition, immunofluorescence analysis is shown in after modeling the 35th day, it is visible bright in control group macrophage
Aobvious STAT3 activation phenomenons, and p-STAT3 signals significantly reduce (Fig. 6 B) after 3-MA processing, illustrate that 3-MA also significantly inhibits huge
The activation of phagocyte STAT3.Finally, we have carried out further verification in RAW cells：3-MA processing can significantly lower by
P-STAT3, total STAT3 and the TGF-β of IL-4 inductions2Expression (Fig. 6 C).Simultaneously we have found that blocking the activation of STAT3
It can inhibit the expression of TGF-β 2, it is meant that TGF-β2Expression may be (Fig. 6 D) of p-STAT3 dependences.
In short, these are the result shows that 3-MA may be by inhibiting the activation of STAT3 in macrophage and TGF-β downstream2
Expression inhibiting fibrosis formation.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, under the premise of the method for the present invention is not departed from, can also make several improvement and supplement, these are improved and supplement also should be regarded as
Protection scope of the present invention.
Claims (9)
- The application of 1.3- methyl adenines in medicine preparation, which is characterized in that the drug is fine under retina for treating Dimensionization.
- 2. application according to claim 1, which is characterized in that the Subretinal Fibrosis refers to age-related macular degeneration phase The Subretinal Fibrosis of pass.
- 3. application according to claim 1, which is characterized in that the formation of the Drug inhibition Subretinal Fibrosis.
- 4. application according to claim 1, which is characterized in that the drug is subtracted by inhibiting macrophage autophagy active Few macrophage quantity, adjusts macrophage subsets, and the activation that macrophage promotees fibrosis signal path is inhibited to play anti-fiber Change acts on.
- 5. application according to claim 1, which is characterized in that the administering mode of the drug is drug administration by injection.
- Application of the 6.3- methyl adenines in the drug for preparing treatment age-related macular degeneration, which is characterized in that the drug suppression The formation of Subretinal Fibrosis processed.
- 7. according to any application of claim 2 or claim 6, which is characterized in that the age-related macular degeneration refers to Moist AMD.
- Application of the 8.3- methyl adenines in reagent preparation, which is characterized in that the reagent is used to adjust macrophage phenotype.
- 9. a kind of pharmaceutical composition of anti-Subretinal Fibrosis, which is characterized in that described pharmaceutical composition includes 3- methyl glands Purine and pharmaceutically acceptable auxiliary material, the Subretinal Fibrosis refer under the relevant retina of age-related macular degeneration Fibrosis, described pharmaceutical composition inhibit the formation of Subretinal Fibrosis.
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| CN114246938A (en) * | 2022-01-25 | 2022-03-29 | 中山大学中山眼科中心 | Use of IL-4 in the manufacture of a medicament for the treatment of retinal degenerative diseases |
| CN114246159A (en) * | 2022-01-04 | 2022-03-29 | 河南省精神病医院(新乡医学院第二附属医院) | Method for preparing neurodevelopmental defect rat model by treating rat with 3-MA (maleic anhydride-maleic anhydride) in weaning period |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114246159A (en) * | 2022-01-04 | 2022-03-29 | 河南省精神病医院(新乡医学院第二附属医院) | Method for preparing neurodevelopmental defect rat model by treating rat with 3-MA (maleic anhydride-maleic anhydride) in weaning period |
| CN114246159B (en) * | 2022-01-04 | 2022-12-06 | 河南省精神病医院(新乡医学院第二附属医院) | Method for preparing neurodevelopmental defect rat model by treating rat with 3-MA (maleic anhydride-maleic anhydride) in weaning period |
| CN114224895A (en) * | 2022-01-06 | 2022-03-25 | 山东农业大学 | Application of 3-methyladenine in removing streptococcus agalactiae in bovine mammary epithelial cells |
| CN114246938A (en) * | 2022-01-25 | 2022-03-29 | 中山大学中山眼科中心 | Use of IL-4 in the manufacture of a medicament for the treatment of retinal degenerative diseases |
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