CN117106028B - Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury - Google Patents
Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury Download PDFInfo
- Publication number
- CN117106028B CN117106028B CN202311198238.5A CN202311198238A CN117106028B CN 117106028 B CN117106028 B CN 117106028B CN 202311198238 A CN202311198238 A CN 202311198238A CN 117106028 B CN117106028 B CN 117106028B
- Authority
- CN
- China
- Prior art keywords
- myocardial ischemia
- protease
- reperfusion injury
- myocardial
- peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 208000031225 myocardial ischemia Diseases 0.000 title claims abstract description 31
- 239000003814 drug Substances 0.000 title claims abstract description 24
- 108091005804 Peptidases Proteins 0.000 title claims abstract description 22
- 239000004365 Protease Substances 0.000 title claims abstract description 22
- 208000007201 Myocardial reperfusion injury Diseases 0.000 title claims abstract description 19
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 15
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title claims abstract 10
- 238000002360 preparation method Methods 0.000 title claims description 5
- 229940079593 drug Drugs 0.000 title abstract description 7
- 210000004027 cell Anatomy 0.000 claims abstract description 31
- 230000002107 myocardial effect Effects 0.000 claims abstract description 30
- 206010021143 Hypoxia Diseases 0.000 claims abstract description 16
- 230000007954 hypoxia Effects 0.000 claims abstract description 16
- 230000036542 oxidative stress Effects 0.000 claims abstract description 16
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 claims abstract description 14
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 claims abstract description 14
- 230000006907 apoptotic process Effects 0.000 claims abstract description 14
- 230000000694 effects Effects 0.000 claims abstract description 13
- 230000010410 reperfusion Effects 0.000 claims abstract description 13
- 102400000069 Activation peptide Human genes 0.000 claims abstract description 9
- 101800001401 Activation peptide Proteins 0.000 claims abstract description 9
- 230000004898 mitochondrial function Effects 0.000 claims abstract description 5
- 210000004413 cardiac myocyte Anatomy 0.000 claims description 18
- 206010019280 Heart failures Diseases 0.000 claims description 7
- 206010048858 Ischaemic cardiomyopathy Diseases 0.000 claims description 7
- 230000000302 ischemic effect Effects 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
- 230000002438 mitochondrial effect Effects 0.000 abstract description 5
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 2
- 239000012528 membrane Substances 0.000 abstract description 2
- 210000001700 mitochondrial membrane Anatomy 0.000 abstract description 2
- 230000009466 transformation Effects 0.000 abstract 1
- 102100039377 28 kDa heat- and acid-stable phosphoprotein Human genes 0.000 description 44
- 101710176122 28 kDa heat- and acid-stable phosphoprotein Proteins 0.000 description 39
- 241000699670 Mus sp. Species 0.000 description 36
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 239000000243 solution Substances 0.000 description 17
- 210000001519 tissue Anatomy 0.000 description 16
- 102000035195 Peptidases Human genes 0.000 description 12
- 230000002861 ventricular Effects 0.000 description 12
- 238000001514 detection method Methods 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 239000000975 dye Substances 0.000 description 9
- 241000700159 Rattus Species 0.000 description 8
- 238000012757 fluorescence staining Methods 0.000 description 8
- 238000007789 sealing Methods 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 238000010186 staining Methods 0.000 description 7
- PLRACCBDVIHHLZ-UHFFFAOYSA-N 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine Chemical compound C1N(C)CCC(C=2C=CC=CC=2)=C1 PLRACCBDVIHHLZ-UHFFFAOYSA-N 0.000 description 5
- 101001135571 Mus musculus Tyrosine-protein phosphatase non-receptor type 2 Proteins 0.000 description 5
- 210000005003 heart tissue Anatomy 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 206010002091 Anaesthesia Diseases 0.000 description 4
- 206010063837 Reperfusion injury Diseases 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 230000037005 anaesthesia Effects 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
- 230000004217 heart function Effects 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000001000 micrograph Methods 0.000 description 4
- 239000011148 porous material Substances 0.000 description 4
- 238000010791 quenching Methods 0.000 description 4
- 230000000171 quenching effect Effects 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 102100025566 Chymotrypsin-like protease CTRL-1 Human genes 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 101000856199 Homo sapiens Chymotrypsin-like protease CTRL-1 Proteins 0.000 description 3
- 206010061216 Infarction Diseases 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 229920004890 Triton X-100 Polymers 0.000 description 3
- 239000013504 Triton X-100 Substances 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000007574 infarction Effects 0.000 description 3
- 210000005240 left ventricle Anatomy 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 208000010125 myocardial infarction Diseases 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 238000010422 painting Methods 0.000 description 3
- 230000035699 permeability Effects 0.000 description 3
- 238000004062 sedimentation Methods 0.000 description 3
- 230000001810 trypsinlike Effects 0.000 description 3
- LNOBZXNCABUBKK-UHFFFAOYSA-N 2,3,5-triphenyltetrazolium Chemical compound C1=CC=CC=C1C(N=[N+]1C=2C=CC=CC=2)=NN1C1=CC=CC=C1 LNOBZXNCABUBKK-UHFFFAOYSA-N 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 102000010825 Actinin Human genes 0.000 description 2
- 108010063503 Actinin Proteins 0.000 description 2
- 238000011740 C57BL/6 mouse Methods 0.000 description 2
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 230000000747 cardiac effect Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000028709 inflammatory response Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 208000028867 ischemia Diseases 0.000 description 2
- 229960002725 isoflurane Drugs 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 210000004165 myocardium Anatomy 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000004080 punching Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000004904 shortening Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000013042 tunel staining Methods 0.000 description 2
- VRYALKFFQXWPIH-PBXRRBTRSA-N (3r,4s,5r)-3,4,5,6-tetrahydroxyhexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)CC=O VRYALKFFQXWPIH-PBXRRBTRSA-N 0.000 description 1
- ZDUOUNIIAGIPSD-UHFFFAOYSA-N 1,1,1-tribromoethane Chemical compound CC(Br)(Br)Br ZDUOUNIIAGIPSD-UHFFFAOYSA-N 0.000 description 1
- CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 description 1
- 230000002407 ATP formation Effects 0.000 description 1
- COXVTLYNGOIATD-HVMBLDELSA-N CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O Chemical compound CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O COXVTLYNGOIATD-HVMBLDELSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 108010067028 Mitochondrial Permeability Transition Pore Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 206010058156 Reperfusion arrhythmia Diseases 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- PMMURAAUARKVCB-UHFFFAOYSA-N alpha-D-ara-dHexp Natural products OCC1OC(O)CC(O)C1O PMMURAAUARKVCB-UHFFFAOYSA-N 0.000 description 1
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000001045 blue dye Substances 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- 230000001964 calcium overload Effects 0.000 description 1
- 238000013155 cardiography Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000009091 contractile dysfunction Effects 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000008694 endothelial dysfunction Effects 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229960003699 evans blue Drugs 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000003601 intercostal effect Effects 0.000 description 1
- 208000037906 ischaemic injury Diseases 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010060 microvascular dysfunction Effects 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 230000004065 mitochondrial dysfunction Effects 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 210000003540 papillary muscle Anatomy 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000037050 permeability transition Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000004845 protein aggregation Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 229940005581 sodium lactate Drugs 0.000 description 1
- 235000011088 sodium lactate Nutrition 0.000 description 1
- 239000001540 sodium lactate Substances 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Cardiology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Hospice & Palliative Care (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a protease activated peptide and application thereof in preparing a medicament for preventing and/or treating myocardial ischemia/reperfusion injury, and relates to the technical field of biological medicines. The amino acid sequence of the protease activation peptide is shown as SEQ ID NO. 1. The invention discovers that the protease activation peptide can reduce the apoptosis after myocardial ischemia/reperfusion and myocardial cell hypoxia/reoxygenation and reduce the oxidative stress level. In addition, the protease activation peptide can also improve the activity of a proteasome, improve the mitochondrial function and state of myocardial cells after hypoxia/reoxygenation, increase the mitochondrial membrane potential, increase ATP generation, reduce membrane permeation and transformation holes and reduce mitochondrial division. Therefore, the protease activation peptide can be used for preparing medicines for preventing and/or treating myocardial ischemia/reperfusion injury.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a protease activated peptide and application thereof in preparing a medicament for preventing and/or treating myocardial ischemia/reperfusion injury.
Background
Ischemic cardiomyopathy refers to left ventricular contractile dysfunction in the case of coronary artery lesions (CAD), and is the most common cause of Heart Failure (HF) worldwide, and ischemic cardiomyopathy patients all have Heart Failure symptoms to some extent. Myocardial ischemia is the most common form of cardiovascular disease with the highest morbidity and mortality. To improve patient prognosis, it is essential to restore timely ischemic myocardium blood flow (reperfusion). However, such reperfusion may lead to further myocardial ischemia/reperfusion injury (MI/RI), resulting in myocardial dysfunction such as myocardial convulsions, reperfusion arrhythmias, muscle cell death, and endothelial and microvascular dysfunction, including no reflux phenomenon, inflammatory response and other myocardial tissue damage, which is more terrible than that caused by the original ischemic injury. Research reports show that the lethal reperfusion injury accounts for 50% of the final myocardial infarction cases. MI/RI pathology involves complex system networks such as oxidative stress, inflammatory response, calcium overload, and mitochondrial dysfunction.
Protease-activated peptides are divided into a closed conformation and an open conformation: PAP and PAP1, the protease-activated peptide significantly increases the chymotrypsin-like (ChT-L) catalytic activity of the proteasome, thereby increasing the rate of proteolysis. In addition, PAP1 culture and H in amyotrophic lateral sclerosis cell model 2 O 2 The stimulated fibroblasts can protect them from death, and PAP1 can prevent protein aggregation. In both cell models, oxidized protein pools decreased after oxidative stress when cells were pre-incubated with PAP 1. PAP1 facilitates the opening of the 20SPT chamber, but has no significant effect on the intracellular polyubiquitinated protein pool, the primary cellular target of PAP1 is the empty 20SPT pool.
However, no report has been made at present on the role of PAP1 in ischemic cardiomyopathy and heart failure.
Disclosure of Invention
The invention aims to provide a protease-activated peptide and application thereof in preparing medicaments for preventing and/or treating myocardial ischemia/reperfusion injury, so as to solve the problems of the prior art, wherein the protease-activated peptide can inhibit myocardial ischemia/reperfusion and apoptosis after myocardial cell hypoxia/reoxygenation, reduce oxidative stress level, improve proteasome activity and improve mitochondrial function and state after myocardial cell hypoxia/reoxygenation, and can be used for preparing medicaments for preventing and/or treating myocardial ischemia/reperfusion injury.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a protease activation peptide, the amino acid sequence of which is shown as SEQ ID NO. 1.
The invention also provides application of the protease activated peptide in preparing medicaments for preventing and/or treating myocardial ischemia/reperfusion injury.
Further, the medicament plays a role in preventing and/or treating myocardial ischemia/reperfusion injury by inhibiting apoptosis after myocardial ischemia/reperfusion and myocardial cell hypoxia/reoxygenation and reducing the level of oxidative stress.
Further, the medicament plays a role in preventing and/or treating myocardial ischemia/reperfusion injury by improving proteasome activity and improving mitochondrial function and state after hypoxia/oxygenation of myocardial cells.
The invention also provides a medicament for preventing and/or treating myocardial ischemia/reperfusion injury, and the active ingredients comprise the protease activated peptide.
Further, the medicament also comprises pharmaceutically acceptable auxiliary materials.
The invention also provides application of the protease activation peptide in preparing a medicament for adjuvant therapy of ischemic cardiomyopathy.
The invention also provides application of the protease activation peptide in preparing medicaments for assisting in treating heart failure caused by ischemic cardiomyopathy.
The invention discloses the following technical effects:
the invention develops a new application of the protease activated peptide PAP1, and researches show that the PAP1 can reduce myocardial ischemia/reperfusion and apoptosis after myocardial cell hypoxia/reoxygenation, and reduce the level of oxidative stress. In addition, PAP1 can also improve proteasome activity, improve mitochondrial function and state after myocardial cell hypoxia/reoxygenation, raise mitochondrial membrane potential, increase ATP production, decrease membrane permeability conversion pores, and decrease mitochondrial division. From this, PAP1 can be used for the preparation of a medicament for preventing and/or treating myocardial ischemia/reperfusion injury. The invention proves that PAP1 can be used for preparing the medicines for auxiliary treatment of ischemic cardiomyopathy and heart failure, and has important clinical guidance significance.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the results of the detection of proteasome activity of rat cardiomyocytes in each experimental group of example 1; wherein A-C are respectively Caspase-like (Caspase-like), trypsin-like (Trypsin-like) and Chymotrypsin-like catalytic activity (Chymotorypsin-like) detection results;
FIG. 2 is a TUNEL fluorescence staining micrograph (A) of rat cardiomyocytes, comparative statistical plot (B) of percent TUNEL positive nuclei for each experimental group of example 1;
FIG. 3 is a comparative chart of DHE fluorescence staining micrographs (A) and relative fluorescence intensities for cardiomyocytes in rats of each experimental group of example 1;
FIG. 4 is a MPTP fluorescent staining micrograph (A) and a fluorescent intensity statistic (B) of cardiomyocytes in rats of each experimental group of example 1;
FIG. 5 shows the results of the heart function test of mice in each experimental group of example 2; wherein A is a heart-comfort moving image; b and C are graphs comparing left ventricular Ejection Fraction (EF) and left ventricular short axis shortening (FS), respectively;
FIG. 6 shows the results of the detection of the heart infarct size in mice of each experimental group of example 2; wherein A is a cross-sectional view of cardiac TTC staining; b and C are the proportion of myocardial dangerous area and infarct area to left ventricular myocardial area respectively;
FIG. 7 is a TUNEL fluorescence staining micrograph (A) of heart tissue of mice of each experimental group of example 2 and a comparative statistical plot (B) of percentage of TUNEL positive nuclei;
FIG. 8 is a microscopic image (A) of DHE fluorescence staining and a statistical image (B) of relative fluorescence intensity of heart tissue of mice of each experimental group of example 2.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Description of the terminology:
the PAP1 is a proteasome activator, is a polypeptide, can be synthesized, and has the structure as follows: ile-Pro-Arg-Cys-Arg-Lys-Met-Pro-Gly-Val-Lys-Met-Cys-NH 2 (SEQ ID NO.1)。
Example 1
1. Neonatal Rat Cardiomyocytes (NRCM) were extracted from 1 day old SD rats and sterilized with 75% alcohol. The left ventricle was removed rapidly with an autoclave-advanced ophthalmic scissors, cut into small pieces, and digested with 0.08% trypsin. The isolated cardiomyocytes were incubated in DMEM/F12 supplemented with 15% fbs for 24 hours and then in serum-free DMEM/F12 for subsequent in vitro studies.
2. Constructing a rat primary myocardial cell hypoxia/reoxygenation model.
NRCM cells were divided into 4 experimental groups, and each experimental group was subjected to the procedure shown in table 1, followed by measurement of proteasome activity and MPTP staining.
TABLE 1 grouping of cell experiments
Experimental group | Operation of |
Vehicle group | NRCM does not perform any operation |
PAP1 group | NRCM was cultured in a medium containing 10. Mu. MPAP1 for 4h |
H/R group | NRCM for in vitro hypoxia/reoxygenation |
PAP1+H/R group | NRCM was cultured in a medium containing 10. Mu. MPAP1 for 4 hours and then subjected to in vitro hypoxia/reoxygenation |
In vitro hypoxia/reoxygenation (H/R) procedure: NRCM containing NaCl and NaHCO 3 、NaH 2 PO 4 ·2H 2 O, anhydrous CaCl 2 、MgCl 2 ·6H 2 Under anoxic conditions in an anoxic buffer of O, sodium lactate, KCl, 2-D-ribose and 2-deoxyglucose (1%O) 2 ) Incubation was performed for 6 hours, and then the cells were incubated in the presence of 10% FBS and 1% penicillin/streptavidinThe plain DMEM/F12 medium is incubated under normal oxygen conditions (95% O 2 ) Reoxygenation was carried out for 24 hours.
3. Proteasome Activity assay
The proteasome activity levels of the 4 groups of primary cardiomyocyte cell suspensions were measured using the protease-GloTM zymo-Trypsin-like, trypsin-like and Caspase-like cell-basedastays (promega, cat No. G8661, G8760. G8861) kit, each of the 4 groups of SD rat milk mouse primary cardiomyocyte (n=6) cell suspensions.
As shown in FIG. 1, the activity of the Chymotrysin-like proteasome of H/R-derived primary cardiomyocytes was decreased as compared to the vector group, and the pretreatment with PAP1 was reversed to increase the proteasome.
TUNEL staining
4 groups of mouse cells were immersed in 20% sucrose in a sink, OCTC-embedded, frozen at-20℃for 20min, baked at 50℃for 10min, fixed with methanol, PBS-washed and stained with tissue on the plates with a tissue brush, perforated with 0.1% Triton X-100 solution at 4℃for 2min, PBS-washed, TUNEL dye incubated at 37℃for 60min, PBS-washed, DAPI-stained for 5min, PBS-washed, α -actinin dye diluted 100-fold with PBS and 30 μl of each tissue overnight at 4 ℃. After rewarming for 30min the next day, diluting 488 fluorescence secondary antibodies with PBS for 100 times, incubating each tissue at 30 mu L and 37 ℃ for 30min, cleaning with PBS solution after the end, sealing the tablet with anti-fluorescence quenching sealing tablet, and performing light-shielding operation in the whole course; fluorescent brightness was photographed with a fluorescent microscope and analyzed using Image J software.
The detection results are as follows: FIG. 2 is a TUNEL fluorescence staining microscopic photograph of the cardiomyocytes of group 4 and a comparative statistical plot of the percentage of nuclei of TUNEL positive cells of group 4, showing increased apoptosis of the cardiomyocytes after I/R; and after the PAP1 pretreatment, the myocardial cell apoptosis is obviously reduced, which indicates that the PAP1 pretreatment can reduce the myocardial cell apoptosis after the myocardial ischemia reperfusion injury of the mice.
Dhe staining
Soaking 4 groups of cells into a sedimentation pond by 20% sucrose, embedding OCTC, freezing and slicing at-20 ℃, baking a slice at 50 ℃ for 20min, fixing the slice for 10min by using methanol, washing by using PBS, circling tissues on the slice by using a painting brush, punching by using 0.5% TritonX-100 solution for 15min, washing by using PBS, diluting DHE dye by 100 times, incubating each tissue for 30min at 30 mu L at 37 ℃, washing by using PBS solution, sealing the slice by using anti-fluorescence quenching sealing tablet after finishing the washing, and performing light-shielding operation in the whole course; fluorescent intensity was photographed with a fluorescent microscope and analyzed using ImageJ software. Primary cardiomyocytes were seeded in 24-well plates, washed with PBS, and fixed in methanol for 10min, the remainder of the procedure was as above.
The detection results are as follows: FIG. 3 is a photograph of a DHE fluorescence staining microscope of myocardial cells of group 4 and a comparative statistical chart of the relative fluorescence intensities of group 4, showing that the myocardial cells have increased oxidative stress levels and increased active oxygen content after I/R; however, the level of oxidative stress in cardiomyocytes was significantly reduced after PAP1 pretreatment, indicating that PAP1 pretreatment was able to reduce oxidative stress after myocardial ischemia reperfusion injury in mice.
MPTP staining
Each group of NRCM cells was MPTP stained as follows: cells were placed in 24 well plates, the culture medium was aspirated, the cells were washed 1-2 times with PBS, caleinAM staining solution was added, and the cells were covered with dye evenly by gentle shaking, and incubated at 37℃for 30min in the absence of light. After the incubation, the culture solution is replaced by fresh culture solution preheated at 37 ℃, and the culture solution is incubated at 37 ℃ for 30 minutes in a dark place, so that intracellular esterase is ensured to fully hydrolyze CalceinAM to generate green fluorescent Calcein. The culture solution was aspirated, washed 3 times with PBS, and then added with detection buffer for observation under a fluorescence microscope.
The detection results are as follows: FIG. 4 is an MPTP fluorescent staining microscope photograph of 4 sets of primary cardiomyocytes and 4 sets of fluorescent intensity statistics, showing that the degree of mitochondrial permeability transition pore opening of H/R post-primary cardiomyocytes is increased; and after the PAP1 pretreatment, the opening degree of the mitochondrial permeability conversion pores of the primary myocardial cells is reduced, which indicates that the PAP1 pretreatment can reduce the opening degree of the mitochondrial permeability conversion pores of the primary myocardial cells after hypoxia reoxygenation.
Example 2
1. Experimental animals and raising
C57BL/6 male mice, 8 weeks old, purchased from Beijing Vitolihua laboratory animal technologies Co., ltd; raising in separate cages, maintaining constant temperature (23-25 deg.c) and constant humidity (55-70%).
2. Experimental grouping
Animal experiments were divided into 4 groups: healthy control group (Vehicle group), normal drug group (PAP 1 group), model group (I/R group) and treatment group (pap1+i/R group), 6 each; as shown in Table 2, the mice in Vehicle group and I/R group were C57BL/6 mice were intraperitoneally injected with 0.2 mL/mouse physiological saline; the PAP1 group and the PAP1+I/R group were each experimental mice pretreated with PAP1 (PAP 110mg/kg, PAP1 was diluted with physiological saline and injected intraperitoneally at an injection amount of 0.2 mL/mouse), and then myocardial ischemia/reperfusion procedures were performed on the I/R group and the PAP1+I/R group, respectively, to evaluate changes in apoptosis and oxidative stress of myocardial tissue, energy metabolism, and related signaling molecules of the mice.
Specific methods for myocardial ischemia/reperfusion procedures are referred to in the references "Y.L.Zhang, P.B.Li, X.Han, B.Zhang, H.H.Li, blockage of fibronectin 1ameliorates myocardial ischemia/reperfusion injury in association with activation of AMP LKB1-AMPK signaling pathway, oxid. Med. Cell. Longev.2022 (2022), 6196173 (1942-0994 (Electronic))".
TABLE 2 grouping of animal experiments
3. Cardiac ultrasound cardiography evaluation
24 hours after reperfusion of the ischemia/reperfusion injury model of the mice, 4 groups of mice are respectively put into an anesthesia box with 1.5% isoflurane for anesthesia, and then are fixed on an operation table in a supine position, and the anesthesia is maintained by 1% isoflurane; firstly, finding a parasternal long axis section, saving B-mode data, rotating a probe for 90 degrees to obtain a parasternal short axis section of a left ventricle of a heart of a mouse, recording left ventricular Ejection Fraction (EF), left ventricular short axis shortening rate (FS), left ventricular end systole inner diameter (ESD), left ventricular end diastole inner diameter (EDD), left ventricular End Systole Volume (ESV) and left ventricular End Diastole Volume (EDV) by using a section of left ventricular papillary muscle level as a mark point on the parasternal short axis section by using an M-type ultrasonic mode; the parasternal long axis section uses the left ventricular outflow tract horizontal section as a mark point, and the B-type ultrasonic cardiogram is used for recording the wall motion image.
The detection results are as follows: the heart-relaxing moving images of the 4 groups of mice recorded by the M-type echocardiogram are shown as A in fig. 5, and the result shows that the heart-relaxing function of the mice after I/R is obviously reduced, and the heart-relaxing function of the model mice pretreated by PAP1 is obviously improved; comparison graphs of EF and FS of 4 groups of mice in sequence in the graphs of B and C in fig. 5 show that EF and FS of model mice are obviously increased after PAP1 pretreatment, and heart function reduction caused by I/R injury can be effectively relieved. The above results show that advanced PAP1 (10 mg/kg) injection into myocardial ischemia reperfusion mice improved cardiac function in the mice.
4.2,3,5-triphenyltetrazolium (TTC) staining
Injecting 0.2mL of tribromoethane supersaturated solution into the abdominal cavity of 4 groups of mice, fixing the mice after anesthesia, ligating the left anterior descending branch of the two groups of mice (Vehicle, PAP 1) which are not molded as in I/R molding, extruding the heart again along the intercostal opening of myocardial molding by the two groups of mice (I/R, PAP1 +I/R) which are molded by the mold, ligating the left anterior descending branch of the heart again at the ligation position of the last time, injecting 0.9mL of 1% Evan blue dye at the apex of the heart, and allowing the dye to flow to the whole body along the blood flow, so that the whole body of the mice can be observed to turn blue; taking down the heart of a mouse, washing the heart with PBS, taking out the wires in the tissue, pouring the glue, putting the heart into a refrigerator at the temperature of minus 20 ℃ for freezing and shaping for 30min, putting the heart into a mould after finishing, cutting the heart into 4 slices with equal thickness, putting the slices into a 1% TTC dye, incubating for 30min in an incubator at the temperature of 37 ℃, putting the heart slices into a 4% tissue fixing solution for fixing for one night after finishing, taking out the heart for photographing the next day.
The detection results are as follows: the cross-sectional view of TTC staining of hearts of 4 mice is shown in fig. 6 a (where blue is a normal region, red is an ischemic region, and white is an infarcted region), the infarcted area of group I/R is significantly increased, and the myocardial infarcted area of pap1+i/R mice is significantly decreased, compared to the Vehicle group; in fig. 6, B and C are the myocardial risk and infarct areas of the 4 groups of mice, respectively, as a proportion of the myocardial area of the left ventricle. The results show that PAP1 pretreatment can partially save myocardial infarction area caused by I/R injury.
TUNEL staining
The hearts of 4 groups of mice are obtained, soaked in 20% sucrose into a sedimentation pond, OCTC embedded, frozen at-20 ℃ for slicing, baked at 50 ℃ for 20min, fixed with methanol for 10min, PBS cleaned and circled out tissues on the slices by a painting brush, perforated with 0.1% Triton X-100 solution at 4 ℃ for 2min, PBS cleaned, TUNEL dye incubated with incubator at 37 ℃ for 60min, PBS cleaned, DAPI dyed for 5min, PBS cleaned, and alpha-actinin dye diluted 100 times with PBS at 30 mu L of each tissue overnight at 4 ℃. After rewarming for 30min the next day, diluting 488 fluorescence secondary antibodies with PBS for 100 times, incubating each tissue at 30 mu L and 37 ℃ for 30min, cleaning with PBS solution after the end, sealing the tablet with anti-fluorescence quenching sealing tablet, and performing light-shielding operation in the whole course; fluorescent brightness was photographed with a fluorescent microscope and analyzed using Image J software.
The detection results are as follows: FIG. 7 is a TUNEL fluorescence staining microscopic photograph of heart tissue of 4 mice and a comparative statistical plot of percentage of nuclei of TUNEL positive cells of 4 mice, showing increased myocardial apoptosis of mice after I/R; and after the PAP1 pretreatment, the apoptosis of the myocardial tissue cells of the mice is obviously reduced, which indicates that the PAP1 pretreatment can reduce the apoptosis of the myocardial cells of the mice after myocardial ischemia/reperfusion injury.
Dhe staining
Drawing the heart of 4 groups of mice, soaking 20% sucrose into a sedimentation pond, embedding OCTC, freezing at-20 ℃ for slicing, baking at 50 ℃ for 20min, fixing with methanol for 10min, washing with PBS, circling tissues on the slices by using a painting brush, punching with 0.5% Triton X-100 solution for 15min, washing with PBS, diluting DHE dye 100 times with PBS, incubating each tissue for 30 mu L at 37 ℃ for 30min, washing with PBS solution, sealing the slices with anti-fluorescence quenching sealing tablets after finishing, and performing whole-course light-shielding operation; fluorescent intensity was photographed with a fluorescent microscope and analyzed using ImageJ software. Primary cardiomyocytes were seeded in 24-well plates, washed with PBS, and fixed in methanol for 10min, the remainder of the procedure was as above.
The detection results are as follows: FIG. 8 is a photograph of a DHE fluorescence staining microscope of heart tissue of 4 mice and a comparative statistical plot of relative fluorescence intensity of 4 mice, showing that the level of oxidative stress in heart tissue of mice after I/R is increased and the active oxygen content is increased; however, the level of oxidative stress in the mouse myocardium decreased significantly after the PAP1 pretreatment, indicating that the PAP1 pretreatment was able to reduce oxidative stress after myocardial ischemia/reperfusion injury in mice.
In summary, the present invention has found that PAP1 increases proteasome activity, reduces apoptosis, reduces oxidative stress level, and increases mitochondrial permeability transition pores after hypoxia/reoxygenation of cardiomyocytes cultured with PAP1 (50. Mu.M). In addition, the invention also establishes a myocardial ischemia/reperfusion model of the mouse, and the PAP1 (10 mg/Kg) is injected in the abdominal cavity for intervention, so that the PAP1 is found to improve the heart function of the mouse after ischemia/reperfusion, reduce the myocardial infarction area of the mouse and reduce the oxidative stress level and apoptosis of myocardial tissues.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (5)
1. The use of a protease-activated peptide, such as Ile-Pro-Arg-Cys-Arg-Lys-Met-Pro-Gly-Val-Lys-Met-Cys-NH, for the preparation of a medicament for the prevention of myocardial ischemia/reperfusion injury 2 As shown.
2. The use according to claim 1, wherein the medicament acts to prevent myocardial ischemia/reperfusion injury by inhibiting apoptosis after myocardial ischemia/reperfusion and myocardial cell hypoxia/reoxygenation and reducing oxidative stress levels.
3. The use according to claim 1, wherein the medicament acts to prevent myocardial ischemia/reperfusion injury by increasing proteasome activity, improving mitochondrial function and status after hypoxia/reoxygenation of cardiomyocytes.
4. Application of protease activated peptide in preparation of medicine for preventing ischemic cardiomyopathyCharacterized in that the protease-activating peptide is, for example, ile-Pro-Arg-Cys-Arg-Lys-Met-Pro-Gly-Val-Lys-Met-Cys-NH 2 As shown.
5. The application of protease activation peptide in preparing medicine for preventing heart failure caused by ischemic cardiomyopathy is characterized in that the protease activation peptide is Ile-Pro-Arg-Cys-Arg-Lys-Met-Pro-Gly-Val-Lys-Met-Cys-NH 2 As shown.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311198238.5A CN117106028B (en) | 2023-09-18 | 2023-09-18 | Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311198238.5A CN117106028B (en) | 2023-09-18 | 2023-09-18 | Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury |
Publications (2)
Publication Number | Publication Date |
---|---|
CN117106028A CN117106028A (en) | 2023-11-24 |
CN117106028B true CN117106028B (en) | 2024-02-13 |
Family
ID=88798373
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311198238.5A Active CN117106028B (en) | 2023-09-18 | 2023-09-18 | Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117106028B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117243950A (en) * | 2023-10-31 | 2023-12-19 | 首都医科大学附属北京朝阳医院 | Application of MK-3903 in preparation of medicines for preventing and/or treating ischemic heart disease |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101511861A (en) * | 2006-07-21 | 2009-08-19 | 克里斯泰利亚化学药物产品有限公司 | Anti-inflammatory and antiallergic cyclic peptides |
-
2023
- 2023-09-18 CN CN202311198238.5A patent/CN117106028B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101511861A (en) * | 2006-07-21 | 2009-08-19 | 克里斯泰利亚化学药物产品有限公司 | Anti-inflammatory and antiallergic cyclic peptides |
Non-Patent Citations (2)
Title |
---|
ACCESSION NO.Q66S08.1.GenBank.2022,全文. * |
Francisco H. Dal Vechio et al..Free Radical Biology and Medicine.2014,第67卷304-313. * |
Also Published As
Publication number | Publication date |
---|---|
CN117106028A (en) | 2023-11-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN117106028B (en) | Protease activated peptide and application thereof in preparation of medicines for preventing and/or treating myocardial ischemia/reperfusion injury | |
US20130236431A1 (en) | Compositions and Methods for Tissue Engineering, Tissue Regeneration and Wound Healing | |
CN106730013A (en) | For preventing Asherman's syndrom and the cell preparation of endometrial impairment reparation and preparation method thereof | |
JP6129418B2 (en) | A composition for preventing and treating liver fibrosis or cirrhosis containing mesenchymal stem cells derived from human embryonic stem cells as an active ingredient | |
JP2021525107A (en) | Cardiomyocyte preparation, its manufacturing method, and application | |
CN109331014A (en) | A kind of purposes of benzimidizole derivatives in preparation treatment heart failure medications | |
WO2021037292A2 (en) | Polypeptide and use thereof | |
CN111298104A (en) | Application of exenatide injection in medicine for treating and preventing intrauterine adhesion | |
CN110483648A (en) | A kind of fused polypeptide and its application | |
US20220347346A1 (en) | Mesenchymal stem cell sheet and use thereof | |
JP5531348B2 (en) | Lower urinary tract disease therapeutic agent and lower urinary tract symptom improving agent | |
CN110934911A (en) | Application of roxburgh rose or extract thereof in preparation of medicine or health-care product for preventing and treating pulmonary fibrosis | |
CN111053800A (en) | Application of blueberries or extracts thereof in preparation of medicines or health-care products for preventing and treating pulmonary fibrosis | |
CN1939453B (en) | Gambir plant extract and its use in preparation of target organ protective medicine | |
TWI590830B (en) | Active ingredients for enhancing stem cell differentiation and used in health care after recovery from myocardial infarction | |
Rosenblatt et al. | Healed subacute bacterial endocarditis | |
CN114107179A (en) | Application of chicken bile exosome in injection medicine for treating chicken liver injury and medicine | |
CN114288301B (en) | Application of DTQ in preparation of medicines for treating acute myocardial infarction and related products | |
CN112426439A (en) | Application of cat adipose-derived mesenchymal stem cells in preparation of preparation for treating cat acute kidney injury | |
CN107184585B (en) | Application of HC-067047 in preparing anti-glioma medicine | |
CN111184743A (en) | Application of multi-type mixed cells in myocardial infarction cell treatment | |
CN100448442C (en) | Use of elastic protease inhibitor for preparing medicine for protecting cerebral hemorrhage | |
CN115227700B (en) | Application of esculentoside A in preparation of myocardial infarction protection medicine | |
CN116270661B (en) | Mitochondrial Na+/Ca2+Application of exchanger blocker in preparation of medicine for improving preeclampsia spiral arterial remodeling failure | |
CN1657101A (en) | Application of liver cells growth factor gene modified myeloid interstitial dry cell in treating mgocardial ischemia |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |