CN105664179A - Method for building animal model with PHF 14 gene knockout-based acute kidney injury and renal fibrosis after injury - Google Patents
Method for building animal model with PHF 14 gene knockout-based acute kidney injury and renal fibrosis after injury Download PDFInfo
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Abstract
The invention relates to the technical field of animal models and concretely, relates to a method for building an animal model with PHF 14 gene knockout-based acute kidney injury and renal fibrosis after injury. The invention builds a model of renal fibrosis after kidney injury under the genetic background of PHF 14 gene knockout under adult conditions and provides essential conditions for researching a use of a PHF 14 gene in fibrotic diseases. Compared with the existing PHF 14 knockout model, the PHF 14 gene knockout provided by the invention does not cause embryonic lethal. A test on adult-stage tamoxifen induced knockout of PHF 14 proves that the PHF 14 gene knockout does not cause embryonic lethal. Compared with a group without knockout, the group subjected to knockout has no substantial pathology of lung, liver and kidney. The method prevents systemic disease state-caused interference on following-up acute kidney injury model making and realizes individual research on use of the gene defect in renal fibrosis pathogenesis.
Description
Technical field
The present invention relates to animal model technical field, specifically, be the method for building up of renal fibrosis animal model after a kind of PHF14 gene knockout merges acute injury of kidney and damages.
Background technology
The Clinical symptoms of acute injury of kidney (acutekidneyinjury, AKI) shows as the renal function sharply decline within a couple of days. Its Etiological is renal ischemic reperfusion injury, and sepsis, nephrotoxic drugs hit and block after kidney. Under AKI mirror, pathology is usually expressed as renal cells necrosis and comes off, and renal tubules intracavity cast is formed, interstitial cell infiltration. Zoopery and Prospective Clinical research all find, even if the kidney short-term renal function that experience AKI hits can recover, proceed to the risk of chronic kidney disease (chronickidneydiseaseCKD) from now on also greater than AKI person did not occur. By AKI proceed to renal interstitial fibrosis be pathological characters CKD specifically mechanism be still not clear, it is possible to after occurring with AKI, local route repair process chronic unbalance and the interaction complicated with the antagonism factor thereof are relevant. This kind of prosthetic process mainly includes cell cycle retardance after injury, the secretion of hypoxia inducible factor, and the activation etc. of TGF-beta signal path. The research worker such as Kitagawa in 2012 find a kind of albumen PHF14 (NM_014660.3) regulating intercellular substance secretion in platelet derived growth factor B alpha (PDGFR-alpha) dependent mode, and have made PHF14 gene unconditional knock-out mice model. Animal model proof PHF14 unconditional knocks out has lethal, and mice can only survive after birth less than 24 hours. The respiratory failure that the pulmonary fibrosis that the cause of the death is serious causes. Postmortem prompting liver and kidney have obvious fibrosis. Experiment in vitro prove PHF14 can the expression of negative regulation PDGFR-alpha, thus suppressing fibroblastic Extracellular Matrix Secretion. (KitagawaM, TakebeA, OnoY, etal.Phf14, anovelregulatorofmesenchymegrowthviaplatelet-derivedgrow thfactor (PDGF) receptor-alpha.TheJournalofbiologicalchemistry.Aug102012;287 (33): 27983-27996.) thus illustrate that PHF14 gene is formed in balance in each organ cell's epimatrix of Embryonic Stages and play important regulative. But also do not illustrate at present in the relation of adult phase Yu fibrosis-related disease about PHF14 gene. In order to study this gene in manhood role in the pathogenic process of renal interstitial fibrosis, it is necessary to a kind of method for building up of Fibrotic animal model after acute injury of kidney and damage under PHF14 gene knockout background, the method currently also has no report.
Summary of the invention
The existing method making PHF14 knock out mice is all unconditional and knocks out, owing to this gene knockout has lethal, so the mouse model that unconditional knocks out cannot be utilized to study the relation of this gene and manhood renal fibrosis. Though and the expression of observable PHF14 gene changes in existing widely used renal fibrosis disease animal model, its effect in disease incidence mechanism cannot be probed into. Therefore be intended to study this gene in manhood role in the pathogenic process of renal interstitial fibrosis, need to set up a kind of animal model knocking out PHF14 gene in manhood condition, Fibrotic research after carrying out acute injury of kidney under this genetic background and damaging. According to experiment needs, folic acid zest nephropathy model can simulate the Clinical Acute injury of kidney course of disease, and after stimulation, renal fibrosis proportion is higher, and can monitor renal function continuously.
It is an object of the invention to set up a kind of method of renal fibrosis animal model after adult PHF14 knocks out the acute injury of kidney simulating clinical disease course under genetic background completely and damages.
Knock out PHF14 according to existing achievement in research known period of embryo whole body and can cause the obvious fibrosis of each major organs, and the manhood after period of embryo's orga-nogenesis knocks out whether this gene has same effect still unknowable. Present invention demonstrates that and after orga-nogenesis, knock out PHF14 without obvious Pathology, it was demonstrated that this gene is not directly pushed into adults in brephic function conclusion. But after the kidney of manhood animal suffers damaging strike, start and have activated a series of Related to repair gene, including PHF14. PHF14 gene function is beyond expression and can cause that renal fibrosis aggravates in which case.
A first aspect of the present invention, it is provided that the method for building up of renal fibrosis animal model after a kind of PHF14 gene knockout merging acute injury of kidney and damage, comprises the following steps:
A, phf14lox+ /+mice and tamoxifen induce Cre to express mice (Cre-ERTM) hybridization, first filial generation screening phf14lox+/-; Cre+ mice;
B, the phf14lox+ that step A is obtained/-; Cre+ mice carries out selfing, second filial filters out phf14lox+ /+; Cre+ mice;
C, the phf14lox+ that step C is obtained /+; Cre+ Mouse feeder, to 8-12 week, accepts continuous 5 days tamoxifen intraperitoneal injections, dosage 3mg/40g body weight, solvent Semen Maydis oil (sigma), the tamoxifen final concentration of 7.5mg/mL in Semen Maydis oil;
D, within the 6th day, give folic acid solution lumbar injection, dosage 250mg/kg body weight, obtain Tubulointerstitial fibrosis animal model after described acute injury of kidney and damage.
Described step D Folic Acid solution is that folic acid is dissolved in 0.3mol/LNaHCO3In, make the final concentration of 10g/L of folic acid. Folic acid is provided by Dalian Mei Lun company. Adopt NaHCO3Being because folic acid as solvent and be insoluble in water, other solvents cannot dissolve. The injection dosage of employing is too small cannot derive acute injury of kidney, and dosage is excessive easily lethal.
The preparation method of described phf14loxp+ /+mice can refer to: HuangQ, ZhangL, WangY, etal.DepletionofPHF14, anovelhistone-bindingproteingene, causesneonatallethalityinmiceduetorespiratoryfailure.Act abiochimicaetbiophysicaSinica.Aug2013;45 (8): 622-633.
Described tamoxifen induces Cre to express mice (Cre-ERTM) provided (B6.Cg-Tg (Cre/Esr1) 5Amc/Jmice (stock004682)) by U.S.'s JAX laboratory.
Described screening phf14lox+/-; Cre+ mice and phf14lox+ /+; The method of Cre+ mice: PHF14 gene and Cre gene are identified respectively, and PHF14 identifies the primer:
PHF14-loxpFa3 '-ctattttcttgattatagatgcag-5 ' (SEQIDNO.1);
PHF14-loxpRa3 '-gccttctaagttccagctactag-5 ' (SEQIDNO.2);
PCR program 95 degree of 1min, 40* (95 degree of 30s, 58 degree of 30s, 72 degree of 1min), 72 degree of 5min, result judges: wt/wt=356bp, wt/flox=356bp+457bp, flox/flox=457bp.
Cre identifies the primer:
CreF3 '-attgctgtcacttggtcgtggc-5 ' (SEQIDNO.3);
CreR3 '-ggaaaatgcttctgtccgtttgc-5 ' (SEQIDNO.4);
PCR program 94 degree of 5min, 35* (94 degree of 20s, 55 degree of 30s, 72 degree of 30s), 72 degree of 5min, result judges: wt is negative. Cre+=200bp.
The second aspect of the invention, it is provided that renal fibrosis animal model after adopting the PHF14 gene knockout that said method is set up to merge acute injury of kidney and damage.
The third aspect of the invention, the method for building up of renal fibrosis animal model after providing above-mentioned PHF14 gene knockout to merge acute injury of kidney and damage, renal fibrosis animal model after the PHF14 gene knockout merging acute injury of kidney set up with said method and damage, the application in preparation or screening acute injury of kidney or the damage Fibrotic medicine of metanephros.
The fourth aspect of the invention, a kind of method screening acute injury of kidney or damage metanephros fibrosis medicine is provided, comprise the following steps: set up renal fibrosis animal model after PHF14 gene knockout merges acute injury of kidney and damages initially with above-mentioned method, then utilize the animal model prepared screening to have the medicine of therapeutic effect for renal fibrosis after acute injury of kidney or damage.
The invention has the advantages that:
1, the present invention establishes fibrosis model after the injury of kidney under the genetic background of a kind of adult condition PHF14 gene knockout, provides necessary requirement for the effect in fibrotic disease of the research PHF14 gene;
2, compared with the existing making PHF14 model knocked out (such as Kitagawa), the PHF14 condition of the present invention knocks out and does not result in embryonic lethal, knock out PHF14 empirical tests in manhood tamoxifen induction and do not result in lethal effect, compared with not knocking out group, knock out group lung, liver, kidney is without notable pathological changes (Fig. 9, Figure 10 and Figure 11), which ensure that follow-up acute injury of kidney modelling is not by the interference of systemic disease state, it is possible to individually research this genetic flaw effect in renal fibrosis pathogenesis.
3, under the genetic background of PHF14 genetic flaw, select folic acid nephropathy to stimulate reason to be the fibrosis course of disease after this model can be simulated Clinical Acute injury of kidney and be damaged as injury of kidney modeling, and renal function can be monitored in different time points; On the other hand, conventional employing unilateral ostruction model then cannot monitor renal function change, also just cannot verify the impact of modelling situation and this gene pairs renal function in renal function level.
Accompanying drawing explanation
Fig. 1. after folic acid stimulates, different time Mouse Blood creatinine FA0d folic acid stimulates 0 day; FA2d folic acid stimulates 2 days;FA14d folic acid stimulates 14 days; FA28d folic acid stimulates 28 days * with FA0d to compare p < 0.05.
Fig. 2. after folic acid stimulates, different time Mouse Blood blood urea nitrogen FA0d folic acid stimulates 0 day; FA2d folic acid stimulates 2 days; FA14d folic acid stimulates 14 days; FA28d folic acid stimulates 28 days * with FA0d to compare p < 0.05.
Fig. 3. folic acid stimulates 0,2,14,28 day kidney PHF14 of mice to express change.
After Fig. 4 .tamoxifen induces PHF14 to knock out, kidney PHF14 down-regulated expression.
The non-knock-out mice of Fig. 5 .PHF14 and PHF14 knock-out mice are at collagen1, alpha-SMA, the TGF-beta expression giving folic acid (FA) different time points afterwards.
Fig. 6 does not give folic acid group Mouse Kidney (A) and gives folic acid 2 (B), 14 (C), and 28 (D) sky kidney Masson dyes.
Fig. 7 .PHF14 does not knock out group (A) and PHF14 knocks out group mice (B) kidney masson pathology, it is seen that two groups of kidneys are all without obvious pathological change.
Fig. 8 .PHF14 does not knock out group (A) and PHF14 knocks out group (B) liver sirius red stains, it is seen that fibrosis zero difference.
Fig. 9 .PHF14 does not knock out group (A) and PHF14 knocks out group (B) lung masson dyeing, it is seen that fibrosis zero difference.
Figure 10 .PHF14 does not knock out group (A) and PHF14 knocks out group (B) and gives kidney masson dyeing during latter 28 days of folic acid (FA).
Figure 11 .PHF14 does not knock out group (A) and PHF14 knocks out group (B) and gives kidney collagen1 SABC during latter 28 days of folic acid (FA).
Figure 12 .PHF14 does not knock out group (A) and PHF14 knocks out group (B) and gives kidney alpha-SMA SABC during latter 28 days of folic acid (FA).
Detailed description of the invention
Below in conjunction with embodiment, detailed description of the invention provided by the invention is elaborated.
Embodiment 1
One, materials and methods
The preparation of 1.AKI animal model and packet: IVC level is grown up C57BL/6 wild-type mice 8～12 week old, body weight 20-35g, provided by U.S.'s JAX laboratory, raise in The 2nd Army Medical College Experimental Animal Center. By random digits table, 20 animals are divided into 4 groups (first-Ding), often group 5. Second-Ding all the animals of group give folic acid and (are dissolved in 0.3mol/LNaHCO3In, make the final concentration of 10g/L of folic acid) lumbar injection, dosage 250mg/kg body weight, folic acid is provided by Dalian Mei Lun company. First treated animal gives 0.3mol/LNaHCO3Lumbar injection, dosage 25mL/kg body weight. First-B-B fraction does not take blood after intraperitoneal administration when 0,2,14 and 28 day, put to death mice, leave and take renal tissue.
The adult specificity induction of 2.PHF14 knocks out model mice and makes: PHF14loxp+ /+mice is given by Chinese Academy of Sciences Shanghai biochemistry Suo Chenzheng army professor, and manufacture method is delivered separately. (HuangQ, ZhangL, WangY, etal.DepletionofPHF14, anovelhistone-bindingproteingene, causesneonatallethalityinmiceduetorespiratoryfailure.Act abiochimicaetbiophysicaSinica.Aug2013; 45 (8): 622-633.) tamoxifen induces Cre to express mice (Cre-ERTM) by U.S.'s JAX laboratory offer (B6.Cg-Tg (Cre/Esr1) 5Amc/Jmice (stock004682)), phf14lox+ /+mice and Cre-ERTMHybridization, first filial generation screening phf14lox+/-; Cre+ mice selfing, second filial filters out phf14lox+ /+; Cre+ mice, raise to 8-12 week, take 8 and be only used as experimental group, separately take same monthly age phf14lox+ /+; Cre-8 is only used as matched group. Experimental group and matched group all accept continuous 5 days tamoxifen intraperitoneal injections, dosage 3mg/40g body weight, solvent Semen Maydis oil (sigma), the tamoxifen final concentration of 7.5mg/mL in Semen Maydis oil.6th day two groups all give folic acid lumbar injection, dosage 250mg/kg body weight. All animals are normally raised afterwards, take blood when the 33rd day, put to death mice, and leave and take kidney, liver and lung tissue.
3. serum creatinine and blood urea nitrogen detection: eye socket blood taking method takes blood 1mL in EDTA coagulant pipe, stands the centrifugal 10min of 4000r/min room temperature after overnight, takes serum to aseptic Eppendorf pipe, and-80 degrees Celsius frozen. Automatic clinical chemistry analyzer measures serum creatinine (SCr) and blood urea nitrogen (BUN).
4. immune-blotting method PHF14, alpha-SMA, the expression of collagen1, TGF-beta: with Tissue lysates (0.1%SDS, 0.5% NaTDC, 150mmol/LNaCl, 50mmol/LTris-HClpH=8.0,0.5% vanadic acid sodium 1%NP-40,1mmol/LNaF, protease inhibitor) the cracking rear 4 degrees Celsius of centrifuging and taking supernatants of tissue, BCA method measures protein concentration. Taking 20 μ g albumen, 8%SDS-PAGE electrophoresis, semidry method goes to pvdf membrane. Primary antibodie is anti-PHF14 (abcam1:1000), GAPDH (SantaCruz1:500), alpha-SMA (abcam1:1000), collagen1 (SAB1:400), ECL reacts, X-ray film exposes, and full automatic gel imaging system is analyzed.
5. histopathological examination: organize 4% paraformaldehyde to fix, paraffin embedding, section, kidney row Masson dyes, alpha-SMA, collagen1 immunohistochemical staining, and lung row Masson dyes, liver row sirius red stains, basis of microscopic observation.
Two, result
1. each group Mouse Blood creatinine and blood urea nitrogen detection: such as table 1, folic acid is injected latter second day and acute injury of kidney is occurred, when within 14 days, entering the chronic fibrosis phase serum creatinine and blood urea nitrogen still relatively baseline have slight rising, 28 days creatinines and blood urea nitrogen then recover further. Different time points serum creatinine (Fig. 1) and blood urea nitrogen (Fig. 2) meet the acute injury of kidney course of disease, acute injury of kidney and chronic fibrosis phase modelling success.
Table 1 folic acid stimulates different time Mouse Kidney function
| SCr(μmol/L) | p | BUN(mg/dL) | P | |
| Folic acid stimulates 0 day | 21.40±0.92 | 11.76±0.28 | ||
| Folic acid stimulates 2 days | 55.58±14.66 | 0.048 | 38.19±10.49 | 0.035 |
| Folic acid stimulates 14 days | 41.46±4.2 | 0.001 | 23.00±2.2 | <0.001 |
| Folic acid stimulates 28 days | 28.04±1.55 | 0.006 | 15.78±1.1 | 0.047 |
Often organize n=5.
2. each group Mouse Kidney PHF14 expression: western blot result shows, compare with lumbar injection solvent group, namely the mice that folic acid stimulates had notable rise from the 2nd day, and continuing high expressed (Fig. 3) in subsequent fiber forming process, the expression of prompting PHF14 is relevant to fibrosis Signal Transduction Pathway Activation.
3.PHF14 clpp gene deratization folic acid stimulate after 14,28 days TGF-beta, collagen1 and alpha-SMA expressions: gene induced knock out after, PHF14 down-regulated expression (Fig. 4). Western blot result shows, after lumbar injection folic acid when 2,14 and 28 days, compares and does not knock out group, TGF-beta, collagen1 and the alpha-SMA up-regulated (Fig. 5) of PHF14 clpp gene deratization.
4. each group kidney of mouse Pathologic changes and ImmunohistochemistryResults Results: visible first group mouse kidney pathological manifestations is normal, and second group mice fibrosis is still inconspicuous, shows as master, Tubular epithelial cell edema, cell infiltration with acute interstitial nephritis. The third, within after the administration of fourth group folic acid 14-28 days, then in the kidney region fibrosis of constantly progress, the shape fibrosis in many stoves, companion's lymphocyte and monocyte infiltration, renal cells atrophy, degeneration, renal tubules tube chamber expands (Fig. 6).
5.PHF14 knock-out mice liver, lung and renal pathology change: kidney masson dyeing display PHF14 knocks out group with the non-group that knocks out all without obvious pathological change (Fig. 7), liver sirius red stains display PHF14 knocks out group with the non-group that knocks out all without obvious pathological change (Fig. 8), and lung masson dyeing display PHF14 knocks out group with the non-group that knocks out also all without obvious pathological change (Fig. 9).To sum up, pathological section is shown in manhood tamoxifen induction and knocks out PHF14 empirical tests and do not result in lethal effect, compared with not knocking out group, knocks out group lung, liver, and kidney is without notable pathological changes.
6.PHF14 knock-out mice renal pathology changes: PHF14 is non-to be knocked out group mice and knocks out group mice kidney masson dyeing display PHF14 when folic acid stimulates 14 days and 28 days and knock out group fibrosis heavier (Figure 10), collagen1 (Figure 11) and alpha-SMA up-regulated (Figure 12), this and westernblot result meet.
Three, discuss
Renal interstitial fibrosis after AKI is the pathologic basis of chronic renal failure. Probing into kidney suffers the AKI mechanism hitting metanephric tubule interstitial fibrosis that the preventing and treating of CKD is significant. The study find that kidney PHF14 gene raises in the AKI model that folic acid causes, continue high expressed and dependence free with Fibrotic progress in the chronic fibrosis phase. This has pointed out the dependency of PHF14 and fibrosis phenotype. The PHF14 manhood induces and shows, during 28 days after suffering AKI strike of the mice knocked out, the renal fibrosis that the wild-type mice relatively suffering same strike is even more serious. Result above prompting PHF14 gene is subject to short fibrosis to stimulate and raise, but its effect is to suppress progression of fibrosis, and after manhood PHF14 gene knockout can cause AKI, fibrosis is aggravated. Find that it plays negative regulation effect in embry ogenesis stage each organ stroma in being formed period of embryo animal about the research of PHF14 gene action before, and prove that its mechanism is that PHF14 protein binding suppresses it to transcribe in platelet derived growth factor B alpha (PDGFR-alpha) transcriptional start site. And then suppress the interstitial mediated by PDGF signal path to be formed. Therefore, each organ PDGFR-alpha over-expression of mice embryonic of PHF14 gene knockout, PDGF signal path excessive activation, interstitial is formed too much. Pathological levels shows as fibrosis. Owing to pulmonary fibrosis ventilatory function is not enough after birth, mice is dead because of respiratory failure. But the work of forefathers is not formulated to this gene of term suffers whether play a role when short fibrosis is hit, the present invention has affirmed this point at kidney.
In the selection of AKI model, we have employed folic acid zest injury of kidney. This is the common model that nephrotoxicity factor causes acute injury of kidney, folic acid stimulates metanephros changes of function to simulate the clinical AKI course of disease completely, and after modeling, fibrosis proportion is higher, and Confounding Factor is few, repeatability is better between animal individual and between modeling batch, and can repeat monitoring renal function. In the animal model of the present invention, we demonstrate PHF14 and resist Fibrotic effect. But the effect of PHF14 in model of blocking after renal ischaemia model and kidney is still worth inquiring into.
For many years, substantial amounts of research has been had to be devoted to illustrate the mechanism of renal fibrosis, although this process relates to the participation of various kinds of cell system and cytokine, generally believe that TGF-beta signal path plays the role of a nucleus in manhood renal fibrosis at present, be Fibrotic initiating and the maintenance factor. Fibroblast can accept TGF-beta stimulation and activate is myofibroblast, and the latter is the main source of extracellular matrix in renal fibrosis. Multiple AKI model all finding, TGF-beta can destroy the integrity of Tubular epithelial cell and promote that epithelial-mesenchymal converts. Inflammation and the immunoreation of local are also had the adjustment effect of complexity and can aggravate the atresia of blood capillary by TGF-beta.In view of finding that the expression of PHF14 and renal fibrosis process have a dependency in our study, and can the generation of negative regulation extracellular matrix. Reasonably speculating it is that TGF-beta have activated the expression of PHF14 by its signal transduction pathway, the PHF14 albumen of increasing expression suppresses the secretion transcribing and then reducing fibroblastic activation and extracellular matrix of PDGFR-alpha. From macroscopically seeing, alleviate the degree of renal interstitial fibrosis. Such ring-type mechanism is then for preventing TGF-beta from excessively urging Fibrotic self limiting mechanism. Should lack Zi limit mechanism or lower and all can promote that manhood kidney is suffering to expedite the progress after AKI hits fibrosis. It is currently known at period of embryo PHF14 by suppressing transcribing of PDGFR-alpha to play the effect suppressing fibrosis phenotype, studies checking further at the whether identical needs of its model of action of manhood. If TGF-beta is elucidated by the PHF14 mechanism formed from limit in progression of fibrosis, then should the potential action site becoming new antifibrosis therapy of target spot on limited signal path.
Below the preferred embodiment of the invention has been illustrated, but the invention is not limited to described embodiment, those of ordinary skill in the art it may also be made that under the premise without prejudice to the invention spirit all equivalent modification or replacement, these equivalent modification or replacement are all contained in the application claim limited range.
Claims (4)
1. a PHF14 gene knockout merges the method for building up of renal fibrosis animal model after acute injury of kidney and damage, it is characterised in that comprise the following steps:
A, phf14lox+ /+mice and tamoxifen induce Cre to express mouse hybrid, and first filial generation screening phf14lox+/-; Cre+ mice;
B, the phf14lox+ that step A is obtained/-; Cre+ mice carries out selfing, second filial filters out phf14lox+ /+; Cre+ mice;
C, the phf14lox+ that step C is obtained /+; Cre+ Mouse feeder, to 8-12 week, accepts continuous 5 days tamoxifen intraperitoneal injections, dosage 3mg/40g body weight, solvent Semen Maydis oil, the tamoxifen final concentration of 7.5mg/mL in Semen Maydis oil;
D, within the 6th day, give folic acid solution lumbar injection, dosage 250mg/kg body weight, obtain Tubulointerstitial fibrosis animal model after described acute injury of kidney and damage.
2. PHF14 gene knockout according to claim 1 merges the method for building up of renal fibrosis animal model after acute injury of kidney and damage, it is characterised in that described step D Folic Acid solution is that folic acid is dissolved in 0.3mol/LNaHCO3In, make the final concentration of 10g/L of folic acid.
3. the method for building up of renal fibrosis animal model after merging acute injury of kidney according to the arbitrary described PHF14 gene knockout of claim 1 or 2 and damage, renal fibrosis animal model after merging acute injury of kidney with the PHF14 gene knockout adopting the arbitrary described method of claim 1 or 2 to set up and damage, the application in preparation or screening acute injury of kidney or the damage Fibrotic medicine of metanephros.
4. the method screening acute injury of kidney or damage metanephros fibrosis medicine, comprise the following steps: set up renal fibrosis animal model after PHF14 gene knockout merges acute injury of kidney and damages initially with the arbitrary described method of claim 1 or 2, then utilize the animal model prepared screening to have the medicine of therapeutic effect for renal fibrosis after acute injury of kidney or damage.
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| CN106689030A (en) * | 2016-11-04 | 2017-05-24 | 南方医科大学 | Application of gene knockout animal as skin barrier function disorder animal model |
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| CN113057142A (en) * | 2021-03-30 | 2021-07-02 | 四川大学华西医院 | Method for constructing intraretinal and/or subretinal fibrosis animal model |
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| CN112136763A (en) * | 2020-09-25 | 2020-12-29 | 四川大学华西医院 | Manba gene knockout renal fibrosis animal model and application thereof |
| WO2022169279A1 (en) * | 2021-02-05 | 2022-08-11 | 연세대학교 산학협력단 | Composition for prevention or treatment of kidney disease |
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