CN105597085A - Application of fibroblast growth factor-21 in treatment of renal fibrosis - Google Patents

Abstract

The present invention discloses application of fibroblast growth factor-21 in treatment and / or prevention of renal fibrosis. The present invention also protects a medicine for treatment and / or prevention of renal fibrosis, and an active ingredient of the medicine is fibroblast growth factor-21. The fibroblast growth factor-21 can effectively inhibit renal fibrosis, can be used as a medicine for treatment and / or prevention of renal fibrosis, and has great value in treatment and prevention of renal fibrosis.

Description

The application of fibroblast growth factor-21 in treatment kidney fibrosis

Technical field

The new purposes of one that the present invention relates to fibroblast growth factor-21, fibroblast growth factor-21 existsApplication in treatment kidney fibrosis.

Background technology

Kidney fibrosis is that various chronic renal diseases advance to jointly lapsing to of end stage renal failure and main pathology basePlinth is one of important diseases of serious harm human health. Kidney region fibrosis is with extracellular matrix (ECM), as collagen I,II, III, IV type, fibronectin, laminin, kidney interstitial too much gather and kidney Stromal fibroblasts hyperplasia isPrincipal character. The progress of prevention and reverse kidney region fibrosis is the key for the treatment of CKD, control renal failure. Kidney interstitial fibreThe pathogenesis of dimensionization is very complicated, comprises inflammatory reaction, response to oxidative stress, Renal Tubular Epithelial Cells Transdifferentiationin etc.

In recent years academia to oxidative stress, the effect in kidney region fibrosis has launched deep research. Oxidative stressReaction is the key link that development occurs kidney region fibrosis, in the overall process that occurs to develop, all rises emphatically at kidney region fibrosisAct on. The fibrillatable initiating stage: the various initiating agents of kidney region fibrosis that can cause are as inflammation, shear stress, hypoxemia, cellThe factor all can affect the intrinsic cell of kidney (renal cells, mesangial cell etc.) and Inflammatory infiltrating cell (huge bitingCell etc.) redox state, stimulate above-mentioned cell to produce a large amount of ROS. Dish fat, nucleic acid etc. on ROS and cell membrane divides greatlySub-material forms lipid peroxidation product (LPO), and the intrinsic cell of coup injury kidney causes kidney cell apoptosis, hypertrophy or turns pointChange, promote kidney region fibrosis to occur; Fibrillatable developing stage: upstream stimulus causes various fibrosis signals to produceAfter, need ROS as important second messenger, transmit and amplify various fibrosis signals. Research shows: ROS can mediateChange the factor (as MCP-1, ICMA-1 etc.) and discharge in a large number, and promote inflammatory cell to assemble at kidney; It is tight that ROS also can induction of vascularPlain II (Angiotenin II, Ang II), transforming growth factor-beta 1 (TransformingGrowthFactor-β,TGF-β) etc. generation the further fibrosis effect in its downstream of mediation of fibrosis key cytokines.

The medicine that lacks clinically at present effective treatment kidney region fibrosis, the measure for the treatment of is mainly to control former diseaseDisease and degradation factors, as hypoglycemic, hypotensive, infection control, tune blood fat, take in high-quality protein diet and improve kidney partMicrocirculation etc. But these modes all can only very limitedly be alleviated advancing of disease, when kidney injury develops into kidney fibrosis can notWhen the reverse stage, patient can only carry out kidney replacement therapy by blood, peritoneal dialysis or kidney transplant and sustain life. ClinicallyMost of CKD patients have occurred seeking medicine treatment after clinical symptoms again, and now kidney has existed fibrillatable in various degreePathology. Therefore inquire into medicine the therapeutic action of established fibrosis lesion is had more to clinical meaning.

Summary of the invention

The object of this invention is to provide the application of fibroblast growth factor-21 in treatment kidney fibrosis.

The present invention also protects a kind of medicine that treats and/or prevents kidney fibrosis, and its active component is that fibroblast is rawLong factor-21. Described medicine also comprises other pharmaceutically acceptable carrier or auxiliary material. In described medicine, except described activityOutside composition, can add the excipient, filler, sorbefacient, surfactant, absorption carrier, the synergy that pharmaceutically allowAgent and additive etc. The administration form of described medicine can be injection (as pulvis, aqua, finish). Described preparation all can adopt thisThose skilled in the art know conventional preparation method and obtain. The method of administration of described medicine can be hypodermic injection, intravenous injectionOr intramuscular injection.

Arbitrary described fibroblast growth factor-21 is following (a) or (b) above:

(a) protein being formed by the amino acid sequence shown in sequence in sequence table 1;

(b) by the amino acid sequence of sequence 1 through the replacement of one or several amino acid residue and/or disappearance and/or interpolation andThere is the protein being derived by sequence 1 of the function that treats and/or prevents kidney fibrosis.

Described fibroblast growth factor-21 is presented as in (c1) to (c7) at least the therapeutic action of kidney fibrosisA kind of:

(c1) suppress kidney fibrosis degree;

(c2) suppressing kidney fibrosis occurs;

(c3) reduce collagenous fibres and/or myofibrillar level in kidney;

(c4) reduce twenty-four-hour urine albumin in kidney, the level of serum creatinine and serum urea nitrogen;

(c5) level of type i collagen albumen and/or α-smooth muscle actin in reduction kidney;

(c6) level of Col IV and/or FN in reduction kidney;

(c7) level of TGF-β and/or phosphorylation Smad2/3 in reduction kidney;

Arbitrary described fibroblast growth factor-21 specifically can adopt the method preparation comprising the steps above:

(1) express the fusion of described fibroblast growth factor-21 and molecular chaperones;

(2) collect described fusion, obtain described fibroblast growth factor-21 after excising described molecular chaperones.

Described molecular chaperones specifically can be molecule ubiquitin sample modified protein (being molecular chaperones SUMO).

The implementation method of described " expressing the fusion of described fibroblast growth factor-21 and molecular chaperones " asUnder: the encoding gene of described fibroblast growth factor-21 is inserted to prokaryotic expression carrier pSUMO, obtain recombinant plasmid; WillDescribed recombinant plasmid imports Escherichia coli Rosetta (DE3), obtains recombinant bacterium; Described recombinant bacterium adopt IPTG to carry out fermentsInduction, obtains described fusion.

The implementation of described " excising described molecular chaperones " is as follows: get described fusion, with SUMO protease IDescribed in (mol ratio of SUMO protease I and fusion specifically can be 1:50) and DTT(, the initial concentration of DTT can be 2mmol/L) cutting (cutting condition specifically can be 4 DEG C of cuttings and spends the night).

The encoding gene of described fibroblast growth factor-21 specifically can be following 1) or 2) or 3) DNA molecular:

1) DNA molecular shown in sequence 2 in sequence table;

2) under stringent condition with 1) the DNA sequence dna hybridization that limits and coding have the DNA molecular of the albumen of identical function;

3) with 1) DNA sequence dna that limits has 90% above homology and coding and has the DNA molecular of the albumen of identical function.

Described stringent condition is at 0.1 × SSPE(or 0.1 × SSC), in the solution of 0.1%SDS, assorted under 65 DEG C of conditionsHand over and wash film.

The present invention finds that fibroblast growth factor-21 can effectively suppress the generation of kidney fibrosis, can be used as medicineTreat and/or prevent kidney fibrosis, there is great value for treatment and the prevention of kidney fibrosis.

Brief description of the drawings

Fig. 1 is the polyacrylate hydrogel electrophoretogram of FGF-21 solution.

Fig. 2 is HE dyeing and Masson coloration result.

Fig. 3 is the superficial density of Masson dyeing.

Fig. 4 is twenty-four-hour urine albumen, serum creatinine and serum urea nitrogen result.

Fig. 5 is Col I gene, a-SMA gene, the table of type i collagen GFP and α-smooth muscle actin gene proteinReach level, the protein level of type i collagen albumen and the relative β-actin of α-smooth muscle actin.

Fig. 6 is, expression and the content of Col IV gene and FN gene.

Fig. 7 is the expression of TGF-β gene, protein level and the phosphorylation of the relative β-actin of TGF-β albumen albumenThe protein level of the relative Smad2/3 albumen of Smad2/3 albumen.

Fig. 8 is the protein level of the relative β-actin of HIPK2.

Detailed description of the invention

Following embodiment is convenient to understand better the present invention, but does not limit the present invention. Experiment in following embodimentMethod, if no special instructions, is conventional method. Test material used in following embodiment, if no special instructions, be fromRoutine biochemistry reagent shop is purchased available. Quantitative test in following examples, all arranges and repeats experiment for three times, and result is made evenAverage.

FGF-21 albumen, be called for short FGF-21, as shown in the sequence 1 of sequence table. The encoding gene of FGF-21 is as sequence tableSequence 2 shown in.

Prokaryotic expression carrier pSUMO(is called as " pHisSUMO " in the literature): bibliography: Jiang Yuanyuan, Yin Chengkai,Li Jinnan, Ren Guiping, Zhang Wei, Li Deshan. utilize the research of the high efficient expression soluble recombinant protein of SUMO emerging system.Northeast Agricultural University's journal, 2008,39 (10): 57-62; Li Lu, Yin Chengkai, Li Deshan. high efficient expression solubility weightHistone expression vector---pHisSUMO. biotechnology, 2009,19 (3): 11-14..

Escherichia coli Rosetta (DE3): Beijing Quanshijin Biotechnology Co., Ltd, catalog number (Cat.No.) CD801.

Male db/db mouse: Shanghai Slac Experimental Animal Co., Ltd., animal quality quality certification SCXK (Shanghai)2015-0005。

PBS buffer solution (pH7.2): solvent is water, and solute and concentration thereof are as follows: NaCl137mmol/L, KCl2.7mmol/L，Na₂HPO₄10mmol/L，KH₂PO₄2mmol/L。

Hematoxylin eosin staining method (hematoxylin-eosinstaining), is called for short HE decoration method: haematine dye liquorFor alkalescence, mainly make endonuclear chromatin and intracytoplasmic ribosomes hyacinthine; Yihong is acid dyes, mainly makes thinComposition red coloration in kytoplasm and extracellular matrix.

Masson dyes (being mainly used in collagenous fibres and myofibrillar differential staining): collagenous fibres are blue (by anilineLan Suoran) or green (being dyed by BG), muscle fibre take on a red color (being dyed by acid fuchsin and Ponceaux).

The preparation of embodiment 1, FGF-21

One, the structure of recombinant plasmid

1, extract total RNA of in vitro people's kidney.

2, the total RNA reverse transcription that adopts M-MLV reverse transcriptase that step 1 is obtained is cDNA.

3, the cDNA obtaining taking step 2 is template, adopts the primer pair of P1 and P2 composition, adopts rTaq enzyme, carries out PCR expansionIncrease, obtain pcr amplification product. Through order-checking, the part between pcr amplification product BsaI and BamHI restriction enzyme site is as sequence tableShown in sequence 2.

P1(underscore mark BsaI recognition site): 5 '-GGGTCTCTAGGTCACCCCATCCCTGACTCCAGT-3′；

P2(underscore mark BamHI recognition site): 5 '-CGCGGATCCTCAGGAAGCGTAGCTGGGGCTTCGG-3。

Pcr amplification program: 95 DEG C of denaturation 5min; 95 DEG C of 30s, 56 DEG C of 30s, 72 DEG C of 45s, 25 circulations; 72 DEG C are prolongedStretch 10min.

4, the pcr amplification product obtaining by restriction enzyme BsaI and BamHI double digestion step 3, reclaims enzyme and cuts product.

5,, with restriction enzyme BsaI and BamHI double digestion prokaryotic expression carrier pSUMO, reclaim the carrier of about 5700bpSkeleton.

6, the carrier framework of the enzyme of step 4 being cut to product and step 5 is connected, and obtains recombinant plasmid pSUMO-FGF-21. HeavyIn group plasmid pSUMO-FGF-21, the coded sequence of the molecular chaperones SUMO on encoding gene and the carrier framework of FGF-21, withAnd the coded sequence (be positioned at the upstream of the coded sequence of SUMO, formed by 6 histidine residues) of His label on carrier frameworkMerge, form fusion, expressed fusion protein (fusion from N hold to C end be followed successively by His label, molecular chaperones SUMO andFGF-21）。

The expection molecular weight of FGF-21 is that 21KD(calculates that molecular weight is 21KD, and SDS-PAGE electrophoresis showed molecular weight is25KD left and right), the expection molecular weight of molecular chaperones SUMO is that 12KD(calculates that molecular weight is 12KD, SDS-PAGE electrophoresis showedMolecular weight is 18KD left and right).

Two, the preparation of FGF-21 and purifying

1, recombinant plasmid pSUMO-FGF-21 is imported to Escherichia coli Rossetta (DE3), obtain recombinant bacterium.

Single colony inoculation of the recombinant bacterium 2, step 1 being obtained to 5mLLB culture medium, the training of vibrating of 37 DEG C, 120rpmSupport 10h, then get bacterium liquid, be inoculated in 500mL containing in the LB culture medium of 50mg/mL penicillin with the volume ratio of 1:100,37 DEG C,120rpm shaken cultivation 2h, now OD_600nmIt is 0.5 left and right.

In the bacterium liquid 3, obtaining to step 2, to add IPTG and make its concentration be 0.25mmol/L inducing (25 DEG C,60rpm shaken cultivation 10h), then 4 DEG C, the centrifugal 30min of 4000rpm, collect thalline.

4, get the thalline that step 3 obtains, carry out ultrasonication (work 1s stops 1s for AMP35%, 8-12min), 4 DEG C,12000rpm is centrifugal, collects respectively supernatant and precipitation.

Respectively supernatant and precipitation are carried out to 12%SDS-PAGE electrophoretic analysis. Result shows, the object egg of solubility expressionAccount in vain the more than 70% of the total destination protein of thalline.

5, get the supernatant that step 4 obtains, carry out HisTrapTMFFcrudecolum affinity chromatography.

Pillar model is: column length 0.7cm, the high 2.5cm of post.

Applied sample amount is 10ml.

Elution process: first use 5 times of column volumes foreign protein eluent (solvent is water, each solute that contains following concentration:40mmol/L imidazoles, 500mmol/LNaCl and 50mmol/LNa₃PO₄; PH7.4) wash-out is to remove foreign protein, and flow velocity is 1ml/Min; Then (solvent is water, each solute that contains following concentration: 500mmol/L to use the destination protein eluent of 3 times of column volumesImidazoles, 500mmol/LNaCl and 50mmol/LNa₃PO₄; PH7.4) wash-out, flow velocity is 1ml/min, 280nm wavelength monitoring is receivedCollection target peak (being the peak of peak value higher than 80mAU), is fusion solution.

6, the fusion solution that adopts HiPrepTM26/10Desalting that step 5 is obtained carries out desalination.

7, get the solution that step 6 obtains, with the mol ratio of SUMO protease I(SUMO protease I and fusion be 1:50) and final concentration be 2mmol/L DTT4 DEG C of cutting spent the night.

8, get the solution that step 7 obtains, carry out HisTrapTMFFcrudecolum affinity chromatography.

Pillar model is: column length 0.7cm, the high 2.5cm of post.

Applied sample amount is 15ml, and 280nm wavelength monitoring is collected target peak (being the peak of peak value higher than 30mAU), is FGF-21Solution.

The polyacrylate hydrogel electrophoretogram of FGF-21 solution is shown in that (swimming lane 1 is molecular weight marker to Fig. 1, and swimming lane 2 is that FGF-21 is moltenLiquid). Reclaim object band and also carry out the order-checking of N end, result shows, N holds front 15 amino acid residues if the sequence 1 of sequence table is from N endHold shown in the 1st to 15 amino acids residues.

Embodiment 2, the FGF-21 therapeutic action to kidney fibrosis.

Animal used as test is the male db/db mouse of cleaning agent in six week age.

The FGF-21 solution that adopts embodiment 1 to prepare is tested.

Animal used as test is divided into 4 groups at random, 6 every group, carries out respectively following parallel processing:

Model control group: high fat high-carbonhydrate diet induction diabetes, the each injection one in every morning 8:00 and abdominal cavity, 20:00 in evening left and right, 2 times/d, totally 9 days, after observation drug withdrawal, within 30 days, there is kidney fibrosis in inferior injection gentamicin 40mg/kg. Test 1-35 days, every10:00 in it morning left and right hypodermic injection PBS buffer solution, single injection volume is 0.2mL;

PFD group: high fat high-carbonhydrate diet induction diabetes, once note of the each injection in every morning 8:00 and abdominal cavity, 20:00 in evening left and rightPenetrate gentamicin 40mg/kg, 2 times/d, totally 9 days, after observation drug withdrawal, within 30 days, occur kidney fibrosis. Test 1-35 days, every the skyNoon 8:00 left and right gavage PFD solution (with PBS buffer solution adjustment medicine), single injection dosage is 500mg/kg body weight, single is filled withStomach injection volume is 0.2mL;

FGF21 group: high fat high-carbonhydrate diet induction diabetes, the each injection in every morning 8:00 and abdominal cavity, 20:00 in evening left and right once, 2 times/d, totally 9 days, after observation drug withdrawal, within 30 days, there is kidney fibrosis in injection gentamicin 40mg/kg. Test 1-35 days, every day8:00 in morning left and right hypodermic injection FGF-21 solution (with PBS buffer solution adjustment protein concentration), single injection dosage is 1.0mgFGF-21(is in total protein)/kg body weight, single injection volume is 0.2mL;

Normal group: test 1-35 days, every morning 8:00 left and right hypodermic injection PBS buffer solution, single injection volume is200ul。

Test the 36th day, mouse is put to death in anesthesia, gets respectively mouse urine, serum and renal tissue.

Collagenous fibres level and muscle fibre level have represented kidney fibrosis degree. Get mouse kidney, make pathological section, pointDo not carry out HE dyeing and Masson dyeing, the results are shown in Figure 2(A is the photo after HE dyeing, and B is the photo after Masson dyeing).Superficial density (superficial density of blue-green collagenous fibres and the red myofibrillar surface after Masson dyeing of Masson dyeingDensity sum) see Fig. 3. Can be seen by Fig. 3: the superficial density of model control group mouse Masson dyeing is very high, i.e. kidneyFibrosis is very high; Compared with model control group, the superficial density of pirfenidone group and FGF21 group mouse Masson dyeingSignificantly reduce, reached the level close with Normal group mouse. Result shows, FGF-21 can significantly suppress kidney fibrosisGeneration.

Classical viewpoint thinks, diabetes cause that extracellular matrix expansion, sertoli cell projection reduce, ceasma diaphragm disappears, endotheliumThe glomerulus pathology such as layer pathology and basement membrane thickened change, with albuminuria close relation. Recently there are some researches show, kidney is littlePipe dysfunction also participates in albuminuria and forms. Regardless of the mechanism of albuminuria, urinary albumin excretion ratio is by extensiveOne of diagnosis index that accreditation is diabetic nephropathy is also the common counter of evaluating result for the treatment of.

Get mouse urine and serum, detect twenty-four-hour urine albumin, kreatinin and urea nitrogen content (adopt urinary albumin,Kreatinin and determination of urea nitrogen kit by specification operation). The unit that the results are shown in Figure the ordinate of 4(Fig. 4 is: every liter of urineIn liquid or serum, contain urinary albumin, the quality of kreatinin and urea nitrogen). Can be seen by Fig. 4: model control group mouse retention is whiteAlbumen, kreatinin and urea nitrogen content are very high, and renal fibrosis degree is very high; Compared with model control group, PFD group andFGF21 group mouse retention albumin, kreatinin and urea nitrogen content significantly reduce. Result shows, FGF-21 can significantly suppress kidneyFibrotic generation.

Type i collagen albumen and α-smooth muscle actin are the marker protein of kidney fibrosis. Get mouse kidney, extract totalRNA reverse transcription are cDNA, detect type i collagen albumen (CollagenI) gene and α-smooth muscle by Real-timePCRThe expression (adopting β-actin gene is reference gene) of actin (α-SMA) gene. For detection of type i collagen albumenThe primer pair of gene is: 5'-GACTGGAAGAGCGGAGAGTACTG-3'; 5'-CCTTGATGGCGTCCAGGTT-3'. For inspectionThe primer pair of surveying α-smooth muscle actin gene is: 5'-CCGAGATCTCACCGAC-3'; 5'-TCCAGAGCTACATGACACAG-3'. Primer pair for detection of reference gene is: 5'-ACATCTGCTGGAAGGTGGAC-3'; 5'-GGTACCACCATGTACCCAGG-3'. Get mouse kidney, extract total protein, detect I type by WesternblotCollagen, α-smooth muscle actin and β-actin protein level (are sheep anti mouse I for detection of the primary antibodie of type i collagen albumenCollagen Type VI antibody, purchased from SantaCruz company; Be the anti-mouse α-smooth muscle of rabbit for detection of the primary antibodie of α-smooth muscle actinActin antibody, purchased from Abcam company). Real-timePCR detects the 5A that the results are shown in Figure of type i collagen GFP.Real-timePCR detects the 5B that the results are shown in Figure of α-smooth muscle actin gene. Fig. 5 C is shown in by the photo of Westernblot. ByFig. 5 can see: model control group mouse type i collagen protein content and α-smooth muscle actin content are very high, i.e. kidneyFibrosis is very high; Compared with model control group, PFD group and FGF21 group mouse hydroxyl type i collagen protein content and α-level and smoothFlesh actin content significantly reduces. Result shows, FGF-21 can significantly suppress the generation of kidney fibrosis.

Get mouse kidney, extracting total RNA reverse transcription is cDNA, detects Col I gene, Col by Real-timePCRIV gene, the expression (adopting β-actin gene is reference gene) of FN gene and a-SMA gene. For detection of Col I baseThe primer pair of cause is: 5'-CGCCATCAAGGTCTACTGC-3'; 5'-GAATCCATCGGTCATGCTCT-3'. For detection ofThe primer pair of Col IV gene is: 5'-AGAAGCGAGATGTTCAAGAAG-3'; 5'-GTTGTGACGGTGGCAGAG-3'. Be used forThe primer pair that detects FN gene is: 5'-CTTTGGCAGTGGTCATTTCAG-3'; 5'-TGGTAGGTCTTCCCATCGTCA-3'.Primer pair for detection of a-SMA gene is: 5'-GACCCAGATTATGTTTGAGACC-3'; 5'-CAGAGTCCAGCACAATACCAG-3', the results are shown in Figure 6.

Get mouse kidney, extracting total RNA reverse transcription is cDNA, detects the table of TGF-β gene by Real-timePCRReach level (adopting β-actin gene is reference gene). Primer pair for detection of TGF-β gene is: 5'-ATGCTAAAGAGGTCACCCGC-3'; 5'-TGCTTCCCGAATGTCTGACG-3'. Get mouse kidney, extract total protein, logicalCross the level that Westernblot detects TGF-β albumen, Smad2/3 albumen, phosphorylation Smad2/3 albumen and β-actin albumen(being rabbit anti-TGF-beta for detection of the primary antibodie of TGF-β albumen, is the anti-Smad2/3 of rabbit for detection of the primary antibodie of Smad2/3 albumen, usesBe the anti-phosphorylation Smad2/3 of rabbit in the primary antibodie that detects phosphorylation Smad2/3 albumen, be all purchased from CellSignalingTechnology company). The results are shown in Figure 7. Real-timePCR detects the 7A that the results are shown in Figure of TGF-β gene. WesternblotPhoto see Fig. 7 B. The photo of Westernblot is carried out to gray scale scanning, obtain the relative β-actin of TGF-β albumen albumenThe protein level of protein level and the relative Smad2/3 albumen of phosphorylation Smad2/3 albumen, the results are shown in Figure 7C. Can be seen by Fig. 7Arrive: model control group mouse TGF-β protein content and phosphorylation Smad2/3 protein content are very high, i.e. renal fibrosis degreeVery high; Compared with model control group, PFD group and FGF21 group mouse TGF-β protein content and phosphorylation Smad2/3 albumen containAmount significantly reduces. Result shows, FGF-21 can significantly suppress the generation of kidney fibrosis.

HIPK2 is conservative serine/threonine core kinases, mainly by transcription factor and the constituent of transcribing original paperPhosphorylation is carried out the expression of regulatory gene. The signal path of HIPK2 mediation TGF-β, and TGF-β is the main of mediation renal fibrosisGene, there are some researches show that HIPK2 is the crucial regulatory factor that promotes renal fibrosis, and therefore HIPK2 is treatment renal fibrosisPotential treatment target spot. Get mouse kidney, extract respectively total protein and nucleoprotein, detect in total protein by WesternblotThe level of HIPK2, detects the level of HIPK2 and β-actin by Westernblot. Figure is shown in by the photo of Westernblot8A. The photo of Westernblot is carried out to gray scale scanning, obtain the protein level of the relative β-actin of HIPK2, the results are shown in Figure8B. Can be seen by Fig. 8: the HIPK2 content in model control group mouse total protein is very high, and renal fibrosis degree veryHigh; Compared with model control group, the HIPK2 content in PFD group and FGF21 group mouse total protein significantly reduces. Result shows,FGF-21 can significantly suppress the generation of kidney fibrosis.

Sequence table

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<120>application of fibroblast growth factor-21 in treatment kidney fibrosis

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Claims (6)

1. fibroblast growth factor-21 treats and/or prevents the application in kidney fibrosis in preparation.

2. application as claimed in claim 1, is characterized in that: described fibroblast growth factor-21 for following (a) or（b）：

(a) protein being formed by the amino acid sequence shown in sequence in sequence table 1;

(b) by the amino acid sequence of sequence 1 through the replacement of one or several amino acid residue and/or disappearance and/or interpolation andThere is the protein being derived by sequence 1 of the function that treats and/or prevents kidney fibrosis.

3. application as claimed in claim 1 or 2, is characterized in that: described fibroblast growth factor-21 is to kidney fibrosisTherapeutic action be presented as at least one in (c1) to (c7):

(c1) suppress kidney fibrosis degree;

(c2) suppressing kidney fibrosis occurs;

(c3) reduce collagenous fibres and/or myofibrillar level in kidney;

(c4) reduce twenty-four-hour urine albumin in kidney, the level of serum creatinine and serum urea nitrogen;

(c5) level of type i collagen albumen and/or α-smooth muscle actin in reduction kidney;

(c6) level of Col IV and/or FN in reduction kidney;

(c7) level of TGF-β and/or phosphorylation Smad2/3 in reduction kidney.

4. treat and/or prevent a medicine for kidney fibrosis, its active component is fibroblast growth factor-21.

5. medicine as claimed in claim 4, is characterized in that: described fibroblast growth factor-21 for following (a) or（b）：

(a) protein being formed by the amino acid sequence shown in sequence in sequence table 1;

(b) by the amino acid sequence of sequence 1 through the replacement of one or several amino acid residue and/or disappearance and/or interpolation andThere is the protein being derived by sequence 1 of the function that treats and/or prevents kidney fibrosis.

6. the medicine as described in claim 4 or 5, is characterized in that: described fibroblast growth factor-21 is to kidney fibrosisTherapeutic action be presented as at least one in (c1) to (c7):

(c1) suppress kidney fibrosis degree;

(c2) suppressing kidney fibrosis occurs;

(c3) reduce collagenous fibres and/or myofibrillar level in kidney;

(c4) reduce twenty-four-hour urine albumin in kidney, the level of serum creatinine and serum urea nitrogen;

(c5) level of type i collagen albumen and/or α-smooth muscle actin in reduction kidney;

(c6) level of Col IV and/or FN in reduction kidney;

(c7) level of TGF-β and/or phosphorylation Smad2/3 in reduction kidney.