CN105079799A - Application of matrix metallo-proteinase inhibitor in resisting nuclear radiation - Google Patents
Application of matrix metallo-proteinase inhibitor in resisting nuclear radiation Download PDFInfo
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Abstract
The invention relates to an application of a matrix metallo-proteinase inhibitor in resisting nuclear radiation, and specifically, the invention relates to biological susceptibility gene mmp9 of nuclear radiation, and further relates to a function for preventing nuclear radiation of a matrix metallo-proteinase inhibitor. The matrix metallo-proteinase inhibitor can be developed into dosage forms including subcutaneous injection, intramuscular injection and intravenous injection, and can be prepared into dosage forms including buccal tablets, nasal sprays, rectum administration agents with transmembrane carriers including transduction peptide and nanometer for prevention and treatment of nuclear radiation, and in addition, the matrix metallo-proteinase inhibitor also can be used for treatment on diseases including tumour.
Description
Technical field
The present invention relates to radioprotective tumor susceptibility gene and matrix metallo-proteinase inhibitor as the development of control radioprotective medicine and the application of matrix metallo-proteinase inhibitor in opposing radioprotective.
Background technology
Peaceful use of nuclear energy facility is extensively promoted in recent years, and nuclear leakage accident frequently occurs.Nuclear weapon exist as military threat always, and the nuclear terrorism utilizing dirty bomb etc. to carry out brings the larger threat of terrorism also to the life of people.While checking severely and being strictly on guard against, preventing trouble before it happens, also must be ready, exploitation antiradiation drug is used for prevention and therapy, loss to be reduced to minimum.
Along with the fast development of Space Science and Technology, manned spaceship goes up to the air in succession, and search plan such as moonfall engineering, mars exploration etc. moves forward steadily, and the outer space medical protection of spaceman has become modern Aviation medical science problem urgently to be resolved hurrily.The outer space, to the mainly complicated ionizing radiation environment of the injury of spaceman, is made up of high flux charged particle and high-octane heavy ion.Heavy ion damage mainly can produce the DNA cluster damage being difficult to repair by inducing cell, has stronger lethal, teratogenesis, mutagenesis and carcinogenesis effect.
Radiation and daily life are also closely related.Along with the develop rapidly of nuclear medicine, doctor contacts radiation chance with patient increases greatly.The safety and Health of the radiation practitioners such as nuclear science worker, radar operator, pilot also attracts people's attention.In fact we move in radiation environment, the environmental radiation such as solar ultraviolet radiation, indoor radon gas, and electromagnetic radiation of mobile phone, TV, computer etc. is ubiquitous.
In sum, along with the develop rapidly of nuclear energy uses, manned space flight and nuclear medicine, the chance that nuclear science worker, spaceman, doctor and patient contact radiation increases greatly, how available protecting and reduce the important topic that radiation damage is medical protection.
Current radiopharmaceutical is all enter the radioelement of human body, scavenging free radicals and short DNA damage reparation etc. based on getting rid of.At present, clinical practice mainly contains WR-2721 (amifostine), superoxide dismutase etc. in the medicine of Antiradiation injury, but toxicity is large, and it is limited that side effect makes it apply more.Find new toxicity low, the antiradiation drug that drug effect is high is problem anxious to be resolved.
Summary of the invention
New there is new mechanism, novel targets, hypotoxic brand-new protective agents to find, inventor utilizes the method for genomics, adopt the technological means such as gene chip, RT-qPCR to find tumor susceptibility gene, and the Antiradiation function of tumor susceptibility gene is verified, and then development antiradiation drug.
The features such as Brachydanio rerio is because of its oophyte and juvenile fish is transparent, be applicable to the high flux screening of medicine, gene and mankind's high conservative have become one of optimal mode biology in life science.Inventor utilizes heavy ion to Brachydanio rerio irradiation, utilizes gene chip and RT-qPCR method to find tumor susceptibility gene mmp9, such as, wherein GELB (MMP9) transcribe or express improve nearly 80 times.The regulator (such as matrix metallo-proteinase inhibitor) that utilization can suppress this tumor susceptibility gene to be expressed carries out radioprotective experiment, finds to have good radiation proof function.
One aspect of the present invention provides the method that whether there is radioprotective in testing environment; it comprises experimental animal is placed in environment certain hour to be detected; then measure transcribing or expression of mmp9 in this experimental animal, and with this experimental animal matrix metalloproteinase in normal circumstances transcribe or compared with expression.
One aspect of the present invention provides and detects animal and whether be subjected to the method for radioprotective, and the method comprises and detects transcribing or expression of mmp9 in this animal, and with the matrix metalloproteinase of this animal under home transcribe or compared with expression.
One aspect of the present invention provides the method for control radioprotective, and it comprises the regulator using transcribing of suppression mmp9 or expression to experimenter.
One aspect of the present invention provides regulator (be preferably matrix metallo-proteinase inhibitor, the be more preferably MMP9 inhibitor) purposes in the medicine for the preparation of control radioprotective of transcribing of suppression mmp9 or expression.
One aspect of the present invention provides the method for the medicine of screening control radioprotective, and whether it comprises mensuration candidate substances can suppress transcribing or expression of mmp9.
One aspect of the present invention provides the preparation of control radioprotective, and it comprises can suppress transcribing or the regulator (be preferably matrix metallo-proteinase inhibitor, be more preferably MMP9 inhibitor) of expression and pharmaceutically acceptable carrier of mmp9.
Accompanying drawing explanation
Fig. 1: carbon ion radiation is to the lethal effect of zebrafish embryo
Note: the embryo of after fertilization 24 hours (24hpf) carries out carbon (weight) ion irradiation of various dose, with dosage increase and the prolongation of time, its mortality rate increases gradually.
Fig. 2: heavy ion irradiation is to the teratogenic effect of Brachydanio rerio
Note: after carbon ion irradiates, at 24h, 48h and 96h equi-time point, observes heavy ion radiation to the morphologic impact of zebrafish embryo.The increasing of irradiation dose, incubation time extends, and deformity is more obvious.There is the phenomenons such as serious rachiocamposis, pericardium and yolk sac edema.
Fig. 3: RT-qPCR screening tumor susceptibility gene
Note: the portion gene RT-qPCR the result of genechip detection difference.
Fig. 4: the determination of matrix metallo-proteinase inhibitor best onset time
Note: during injection Ilomastat 2h, the expression of MMP9 is minimum, is best onset point when 2h is described.
Fig. 5 .1-Fig. 5 .3: the radiation-resisting functional research of matrix metallo-proteinase inhibitor (Ilomastat);
The different exposure dose of Fig. 5 .1 is on the impact of zebrafish embryo abnormal rate.
Note: give Ilomastat 0.5mg/ml (10ng/ fish, administration group) and compare with the matched group not giving Ilomastat, the deformity performance after irradiation obviously alleviates, and abnormal rate obviously reduces.
The different exposure dose of Fig. 5 .2 is on the impact of zebrafish embryo mortality rate
Note: give Ilomastat 0.5mg/ml (3-8ng/ fish, administration group) and compare with the matched group not giving Ilomastat, the mortality rate of administration group is starkly lower than matched group.
The different exposure dose of Fig. 5 .3 is on the impact of zebrafish embryo deformity performance
Note: give Ilomastat 0.5mg/ml (3-8ng/ fish, administration group) and compare with the matched group not giving Ilomastat, the deformity performance of administration group is starkly lower than matched group.
Fig. 6: matrix metallo-proteinase inhibitor (Ilomastat) is to Brachydanio rerio MMP9 Gene Expression
Note: the administration group of 15Gy exposure dose obviously reduces than the expression of non-administration group MMP9, illustrates that the expression of Ilomastat to MMP9 gene is inhibited.
Detailed description of the invention
In the present invention, " environment " wherein comprises any physical space of the mankind or other biological survival activity, and it includes but not limited in the outer space, atmosphere, soil or water.Specifically, environment to be detected can be space station, spacecraft, aerospace detection device, troposphere, stratosphere, intermediate layer, thermosphere and loss layer, building, fresh water or marine park, river, shallow sea or neritic zone, abysmal area, mine, drilling well, farmland, pasture, high mountain, hills, streams etc.
In the present invention, the animal that described " experimental animal " selects its function, metabolism, structure and disease character similar to people as far as possible.As a rule, if the phylogenetic scale residing for selected animal is higher, the result of the test so obtained by it is also more meaningful to we mankind.Such as, the non-human primate such as orangutan, macaque, baboon are similar to the mankind most, has reference especially according to the practice of result to the mankind that the present invention especially obtains.Further, " experimental animal " of the present invention includes but not limited to: mice, rat, Cavia porcellus, Carnis Coturnicis japonicae, pig, horse, cattle, sheep, chicken, duck, goose, Columba livia, rabbit, fish (Carassius auratus, Misgurni anguillicaudati etc.), dog, cat, Rana nigromaculata, Bufo siccus, newt, terrapin, Carnis Mactrae, Eriocheir sinensis class, echinoid, fly class, mosquito class, Blatta seu periplaneta, birds, Rodents (as Cricetulus barabensis), primates (orangutan, macaque, baboon) etc.
In the present invention, time experimental animal being placed in environment to be detected can be 1 minute to 6 months or any time section in 60 middle of the month.Such as, time experimental animal being placed in environment to be detected can be 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60 minute; 11,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60 hours; 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60 days; Or 1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60 months.
In the present invention in determination test animal mmp9 transcribe or the method for expression can adopt various method well known in the prior art, include but not limited to: real-time quantitative PCR (RT-qPCR), Northernblotting, Western blotting or adopt reporter gene or merge the direct measuring method of green fluorescent protein, beta galactosidase or luciferase means.
In determination test animal, mmp9's transcribes or after expression, if the described gene of experimental animal has significant difference with expression in normal circumstances in environment to be measured, then indicates in this environment to be measured and there is radioprotective.If the described gene of experimental animal does not exist significant difference with expression in normal circumstances in environment to be measured, then indicate in this environment to be measured and there is not radioprotective.
In this article, " significant difference " refers to that the expression such as in environment to be measured described in correlation test animal is more than 1.2 times or less than 0.8 times of the expression under home, such as, be 1.2-200 times or 0.8-0.0 times.Exemplarily, described expression can be such as 1.3,1.4,1.5,1.6,1.7,1.8,1.9,2.0,2.1,2.2,2.3,2.4,2.5,2.6,2.7,2.8,2.9,3.0,3.1,3.2,3.3,3.4,3.5,3.6,3.7,3.8,3.9,4.0,4.1,4.2,4.3,4.4,4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8,5.9 or 6.0 times; 7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60,70,80,90,100,110,120,130,140,150,160,170,180,190 or 200 times.Or, described in be expressed as 0.8,0.7,0.75,0.6,0.65,0.5,0.55,0.4,0.45,0.3,0.35,0.2,0.25,0.1,0.15,0.05 or 0.0 times.
In this article, " significant difference " comprises " being significantly higher than ".Such as, " being significantly higher than " refers to, such as in environment to be measured, the expression of the gene (such as mmp9) of correlation test animal is more than 1.2 times of the expression under home, such as, be 1.2-200 times.Exemplarily, described expression can be such as 1.3,1.4,1.5,1.6,1.7,1.8,1.9,2.0,2.1,2.2,2.3,2.4,2.5,2.6,2.7,2.8,2.9,3.0,3.1,3.2,3.3,3.4,3.5,3.6,3.7,3.8,3.9,4.0,4.1,4.2,4.3,4.4,4.5,4.6,4.7,4.8,4.9,5.0,5.1,5.2,5.3,5.4,5.5,5.6,5.7,5.8,5.9 or 6.0 times; 7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59 or 60,70,80,90,100,110,120,130,140,150,160,170,180,190 or 200 times.
One aspect of the present invention provides and detects animal and whether be subjected to the method for radioprotective, and the method comprises and detects transcribing or expression of mmp9 in this animal, and with the matrix metalloproteinase of this animal under home transcribe or compared with expression.
Measure gene described in animal subject transcribe or after expression; if the expression of the described gene of relevant animal subject and mutually homozoic expression in normal circumstances exist significant difference, then indicate this animal subject encountered or meet with radioprotective.If the gene expression dose of relevant animal subject and mutually homozoic expression in normal circumstances do not exist significant difference, then the non-encountered of this animal subject is indicated to cross or do not meet with radioprotective.
In this article, " regulator " can refer to suppress or significantly suppress transcribing or any material of expression of mmp9.
In this article, described " adjustment " nonrestrictive expression comprising " suppression " related gene.Such as can regulate transcribing or expression of (suppression) mmp9 by significance difference different in naturely.More specifically, described regulator energy significance difference suppresses transcribing of mmp9 or expression different in naturely.In this article, described " suppression " can refer to, such as, the expression of related gene after suppressing be before suppressing transcribe or expression be 0.8,0.7,0.75,0.6,0.65,0.5,0.55,0.4,0.45,0.3,0.35,0.2,0.25,0.1,0.15,0.05 or 0.0 times.
Matrix metallo-proteinase inhibitor described in the present invention can be any suppression or any inhibitor eliminating matrix metalloproteinase (MMP, preferred MMP9), and this kind of inhibitor can be known to those skilled in the art.Exemplary MMP inhibitor is as described below.Hill, P.A.etal., BiochemJ (1995) 308:167-175 describes two kinds of MMP inhibitor: CT1166 and RO317467.Gowravaram, M.R.etal., JMedChem (1995) 38:2570-2581 describes and suppresses a series of hydroxamate of MMP and be referred to as the thio-alcohol of MMP inhibitor, phosphonic acid based, phosphonous acid class, phosphoramidic acid esters and N-carbonylic alkyl class.This paper is pointed out, MMP inhibitor packages is containing the part of sequestration zinc and the fragments of peptides in conjunction with MMP specificity pockets subgroup.Hodgson, J., Biotechnology (1995) 13:554-557 this section of summary has addressed the clinical condition of several MMP inhibitor, comprises Galardin, Batimastat and Marimastat.Other MMP inhibitor comprise succinamide (Conway, J.G.etal., JExpMed (1995) 182:449-457), TIMP (MauchC., etal., ArchDermatolRes (1994) 287:107-114) and V-A acidic (Fanjul, A.etal., Nature (1994) 372:107-111; Nicholson, R.C.etal., EMBOJournal (1990) 9 (13) 4443-4454; AndBailly, C.etal., JInvestigDerm (1990) 94 (1): 47-51).Matrix metallo-proteinase inhibitor also comprises succinyl hydroxamic acid class matrix metallo-proteinase inhibitor (such as Yi Luo Maas he).
Another aspect of the invention provides a kind of pharmaceutical composition or pharmaceutical preparation, and it comprises gene expression regulator of the present invention, such as matrix metallo-proteinase inhibitor.Preferably, it is further containing being selected from following adjuvant: binding agent, disintegrating agent, diluent, lubricant, fluidizer, emulsifying-solubilizing agent, sweeting agent, coating material and antibiotic antiseptic, such as Sulfobutyl ether β _ cyclodextrin and Polyethylene Glycol.For pharmaceutical composition of the present invention or pharmaceutical preparation, it can be used for preventing and treating radioprotective.
For pharmaceutical composition of the present invention or pharmaceutical preparation, it is used by injection, sublingual administration, lung suction, nasal mucosa and the various ways such as anus bolt or skin absorption, eye drop.Its application dosage is preferably 20mgMMP/kg body weight-45mgMMP/kg body weight.
Of the present invention comprise gene expression regulator (such as matrix metallo-proteinase inhibitor) pharmaceutical composition or pharmaceutical preparation can be developed into the dosage forms such as subcutaneous injection, intramuscular injection, intravenous injection, and make buccal tablet, the dosage form such as nasal spray, rectum feed.Described pharmaceutical composition or pharmaceutical preparation can be used for the prevention and therapy of radioprotective.
Below will by illustrating in greater detail the present invention by following examples.Following examples are only illustrative, should be understood that the present invention not by the restriction of following embodiment.
Embodiment 1. heavy ion radiation is on the impact of Zebrafish Embryo
Inventor carries out experimentation with conventional heavy ion (carbon particle) in radioprotective experiment, utilizes the heavy ion of various dose to carry out radiation to zebrafish embryo, observes afterwards, calculate mortality rate, abnormal rate and lopsided phenotype.Specific experiment is carried out as follows: join fish in a few days ago evening of experiment, morning pumps dividing plate, and male, raun is knocked into the back, is fertilized.Collect same batch of germ cell.Wash Brachydanio rerio germ cell 3-4 time with E3 culture fluid, wash away the Excreta of Brachydanio rerio, foodstuff and other impurity, afterwards germ cell is put into 28 DEG C of constant incubators and hatch.In the process of hatching, Timeliness coverage is removed the dead ovum of unfertilized egg and albefaction and is changed water.After cultivating 24h, the embryo of same Parent is evenly distributed in 6 orifice plates, every hole 20-30 piece embryo, divides well and carry out irradiation respectively.This experiment uses 12C6+ proton, and close rate is 7.5Gy/min, and irradiation dose is respectively 3Gy, 6Gy, 9Gy, 12Gy and 15Gy.An exposure dose arranges 3 repeating groups, establish blank group (not carrying out irradiation) simultaneously, after heavy ion radiation, timing basis of microscopic observation also calculates the mortality rate (Fig. 1 shown in) of zebrafish embryo, abnormal rate and lopsided phenotype (shown in Fig. 2) etc.Adopt SPSS17.0 software to carry out analysis of experimental data, use GraphPad software to carry out mapping analysis.Found that, the heavy ion radiation of various dose has teratogenesis, the effect such as lethal to zebrafish embryo.
Embodiment 2 heavy ion radiation tumor susceptibility gene finds
The zebrafish embryo coming from same Parent after fertilization 24h is placed in 6 orifice plates, 20-30 piece of embryo is put in every hole, be divided into 12 holes, be divided into 9 radiation groups and 3 matched groups (non-irradiated group), irradiation group is carried out the 12C6+ proton irradiation of 9Gy, 12Gy, 15Gy, and continue to cultivate, hatch, after irradiation 7d, the Brachydanio rerio of irradiation group and Normal group is blotted water be put in cryopreservation tube and freeze in liquid nitrogen, for follow-up genechip detection.
Chip synthesis, signals collecting, quality control and analysis are undertaken by Boao Biological Co., Ltd's Beijing National Engineering Research Center.Data analysis all adopts the MAS3.0 system of Bo Ao company.According to the analysis result of gene chip, find the gene that diversity is larger, design primer according to its transcript number by the online website of VizPrimer (www.biocompute.bmi.ac.cn//CZIab/VizPrimer/ /).Primer is synthesized by match Parkson, Beijing gene technology company limited.
RT-qPCR experimental verification is carried out to the gene differed greatly.The reagent, centrifuge, super-clean bench etc. of extraction RNA all need the DEPC water wiping with 0.1%, and the rifle head used in leaching process is the rifle head without RNA enzyme.RNA extracts daily root total RNA extraction reagent box description to carry out (operating procedure slightly).The multi-functional microplate reader of the RNA extracted measures the OD value at 260nm and 280nm wavelength place, pure RNAOD260/OD280 ≈ 2.0 (this experiment is between 1.8 ~ 2.0).
Reverse transcription uses PromegaRNA Reverse Transcription box, and step by specification carries out.The cDNA obtained dilutes 5 times, is sub-packed in the EP pipe of 1.5mL, and-20 DEG C of preservations are for subsequent use.
RT-qPCR is with β-actin for reference gene, and its design of primers is as follows:
Forward primer: 5 '-CTCAGGATGCGGAAACTGGC-3 '
Downstream primer: 5 '-CATGGACGCCCATTGTGAGG-3 '
Each sample sets 3 repetitions.Above action need in clean super-clean bench lucifuge and need operate on ice.After adding sample, need EP pipe lid to cover tightly, then of short duration centrifugal, be put into the amplification and real-time monitoring of carrying out PCR in fluorescence real-time quantitative PCR instrument.Adopt SPSS17.0 software to carry out analysis of experimental data, use GraphPad software to carry out mapping analysis.Found that, heavy ion radiation can relate to many signal paths, and it is comparatively large that relevant signal path proportion wherein occurs to tumor, such as TGF, Wnt, p53 path etc.Wherein have chosen some genes, carry out RT-qPCR checking, find that expression change in various degree appears in mmp9 gene, as shown in Figure 3.
The antiradiation effect of embodiment 3 matrix metallo-proteinase inhibitor
Zebrafish embryo carries out heavy ion radiation after giving matrix metallo-proteinase inhibitor, calculates its mortality rate, incubation rate and lopsided phenotype, thus draws radioprotective effect.The representational medicine of matrix metallo-proteinase inhibitor most be Yi Luo Maas he (Ilomastat), inventor with him for experimental drug carries out series of studies.Specific experiment step: join fish, morning pumping board first 2 day evening in heavy ion irradiation test, male, raun is knocked into the back, is fertilized.Collect Brachydanio rerio germ cell, clean, be placed in 28 DEG C of incubators and hatch.Inject after 24hpf, the matrix metallo-proteinase inhibitor solution 10nl of every piece of embryo's injection 0.5mg/ml, injecting normal saline is matched group.The determination of best onset time, after drug administration by injection, at 0.5,1,2,4,8 hour etc., each time point extracted embryo's total serum IgE, reverse transcription becomes cDNA, carry out RT-qPCR detection, being detected object with GELB, is the best onset time that matrix metallo-proteinase inhibitor suppresses MMP9 when lowering minimum.This laboratory observation 2 hours is the best onset time of matrix metallo-proteinase inhibitor to administration, shown in Fig. 4.So be all that administration carries out radiation in 2 hours to the radioprotective research of matrix metallo-proteinase inhibitor.Within after drug administration by injection 2 hours, embryo is carried out to the heavy ions of the various dose such as 3Gy, 6Gy, 9Gy, 12Gy, 15Gy, the matched group of injecting normal saline, also the heavy ions of the various dose such as 3Gy, 6Gy, 9Gy, 12Gy, 15Gy is carried out respectively, exposure dose rate is 7.5Gy/min, administration group and non-administration group are often organized and are all established 3 parallel repetitions, have eachly repeated 20-30 zebrafish embryo.Observe and after calculating drug effect radiation to the incubation rate of Brachydanio rerio, mortality rate and phenotype etc.After cultivating 7d, extract Brachydanio rerio RNA, carry out according to the total RNA extraction reagent box description of the biological company limited of sky, Beijing root.Carry out reverse transcription immediately after extraction, the cDNA after reverse transcription be kept at-80 DEG C for subsequent use.Quantitative fluorescent PCR (RT-qPCR), condition system is all identical with embodiment 2.Whether observe medicine can be inhibited to MMP9 gene expression.Adopt SPSS17.0 software to carry out analysis of experimental data, use GraphPad software to carry out mapping analysis.Result display administration group abnormal rate obviously declines (shown in Fig. 5 .1), and mortality rate obviously reduces (shown in Fig. 5 .2), and deformity performance obviously alleviates (shown in Fig. 5 .3).Radiation still can make MMP9 gene expression raise, but by nearly 80 times that not administration is raised, only raises 10 times (shown in Fig. 6) after administration.Matrix metallo-proteinase inhibitor (Yi Luo Maas he) makes the rise ratio of MMP9 gene have dropped about 8 times.
Although with above embodiments describing the present invention, should be understood that, under the prerequisite not deviating from spirit of the present invention, the present invention can further modify and change, and these are modified and variation all belongs within protection scope of the present invention.
Claims (10)
1. in testing environment, whether there is the method for radioprotective; it comprises experimental animal is placed in environment certain hour to be detected; then measure transcribing or expression of this experimental animal matrix metalloproteinase, and with this experimental animal described gene in normal circumstances transcribe or compared with expression.
2. method according to claim 1, wherein said matrix metalloproteinase is GELB.
3. detect animal and whether be subjected to the method for radioprotective, the method comprises and detects transcribing or expression of this animal matrix metalloproteinase, and with the described gene of this animal under home transcribe or compared with expression.
4. method according to claim 3, wherein said matrix metalloproteinase is GELB.
5. the method for the medicine of screening control radioprotective, whether it comprises mensuration candidate substances can suppress transcribing or expression of matrix metalloproteinase.
6. method according to claim 5, wherein said matrix metalloproteinase is GELB.
7. whether method according to claim 5, wherein measure candidate substances and the method for transcribing of matrix metalloproteinase or expression can be suppressed to comprise RT-qPCR, Northernblotting, Western blotting or adopt the direct measuring method of reporter gene or fusion green fluorescent protein, beta galactosidase or luciferase means.
8. can suppress matrix metalloproteinase (preferred mmp9) transcribe or expression regulator (inhibitor of such as matrix metalloproteinase, preferred substrate MMP-9 inhibitor) for the preparation of control radioprotective medicine in purposes.
9. purposes according to claim 8, wherein said matrix metalloproteinase is GELB.
10. the purposes of according to Claim 8 or 9, wherein said matrix metallo-proteinase inhibitor is succinyl hydroxamic acid class (such as he and analog thereof of Yi Luo Maas).
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| CN108113996A (en) * | 2016-11-29 | 2018-06-05 | 中国人民解放军军事医学科学院毒物药物研究所 | A kind of water-soluble skin protective agent protected for chemical poison and its preparation method and application |
| CN113304108A (en) * | 2021-05-24 | 2021-08-27 | 中国人民解放军军事科学院军事医学研究院 | Ilomastat suspension injection and preparation method thereof |
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| CN108113996A (en) * | 2016-11-29 | 2018-06-05 | 中国人民解放军军事医学科学院毒物药物研究所 | A kind of water-soluble skin protective agent protected for chemical poison and its preparation method and application |
| CN113304108A (en) * | 2021-05-24 | 2021-08-27 | 中国人民解放军军事科学院军事医学研究院 | Ilomastat suspension injection and preparation method thereof |
| CN113304108B (en) * | 2021-05-24 | 2022-03-15 | 中国人民解放军军事科学院军事医学研究院 | Ilomastat suspension injection and preparation method thereof |
Also Published As
| Publication number | Publication date |
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| CN111840556B (en) | 2023-06-02 |
| CN111840556A (en) | 2020-10-30 |
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