CN107693784B - Pedf基因在治疗糖尿病性心肌损伤中的应用 - Google Patents
Pedf基因在治疗糖尿病性心肌损伤中的应用 Download PDFInfo
- Publication number
- CN107693784B CN107693784B CN201710819105.3A CN201710819105A CN107693784B CN 107693784 B CN107693784 B CN 107693784B CN 201710819105 A CN201710819105 A CN 201710819105A CN 107693784 B CN107693784 B CN 107693784B
- Authority
- CN
- China
- Prior art keywords
- pedf
- gene
- myocardial
- diabetic
- mice
- 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
- 108090000102 pigment epithelium-derived factor Proteins 0.000 title claims abstract description 72
- 206010012601 diabetes mellitus Diseases 0.000 title claims abstract description 57
- 208000037891 myocardial injury Diseases 0.000 title claims abstract description 27
- 238000011282 treatment Methods 0.000 title description 7
- 102100035846 Pigment epithelium-derived factor Human genes 0.000 claims abstract description 57
- 239000003814 drug Substances 0.000 claims abstract description 18
- 229940079593 drug Drugs 0.000 claims abstract description 6
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 6
- 230000004064 dysfunction Effects 0.000 claims abstract description 5
- 238000001415 gene therapy Methods 0.000 claims abstract description 5
- 239000003596 drug target Substances 0.000 claims abstract description 4
- 238000012216 screening Methods 0.000 claims abstract description 4
- 206010007572 Cardiac hypertrophy Diseases 0.000 claims description 9
- 230000000747 cardiac effect Effects 0.000 claims description 9
- 206010028594 Myocardial fibrosis Diseases 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 3
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 2
- 239000012190 activator Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims 1
- 230000003680 myocardial damage Effects 0.000 abstract description 18
- 230000006870 function Effects 0.000 abstract description 12
- 210000004369 blood Anatomy 0.000 abstract description 11
- 239000008280 blood Substances 0.000 abstract description 11
- 230000002107 myocardial effect Effects 0.000 abstract description 7
- 230000004217 heart function Effects 0.000 abstract description 6
- 206010016654 Fibrosis Diseases 0.000 abstract description 4
- 230000004761 fibrosis Effects 0.000 abstract description 4
- 206010020880 Hypertrophy Diseases 0.000 abstract description 3
- 239000003124 biologic agent Substances 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 3
- 238000003209 gene knockout Methods 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 41
- 241000699670 Mus sp. Species 0.000 description 31
- 241000699666 Mus <mouse, genus> Species 0.000 description 21
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000008096 xylene Substances 0.000 description 10
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 8
- 238000011813 knockout mouse model Methods 0.000 description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 206010019280 Heart failures Diseases 0.000 description 6
- 230000017531 blood circulation Effects 0.000 description 6
- 239000012153 distilled water Substances 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 239000012188 paraffin wax Substances 0.000 description 6
- 230000001575 pathological effect Effects 0.000 description 6
- 208000008589 Obesity Diseases 0.000 description 5
- 210000000577 adipose tissue Anatomy 0.000 description 5
- 230000037396 body weight Effects 0.000 description 5
- 230000003205 diastolic effect Effects 0.000 description 5
- 235000020824 obesity Nutrition 0.000 description 5
- 238000004904 shortening Methods 0.000 description 5
- 208000032781 Diabetic cardiomyopathy Diseases 0.000 description 4
- 206010052337 Diastolic dysfunction Diseases 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 201000010063 epididymitis Diseases 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 235000021588 free fatty acids Nutrition 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 210000002303 tibia Anatomy 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 208000006029 Cardiomegaly Diseases 0.000 description 3
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 210000004413 cardiac myocyte Anatomy 0.000 description 3
- 210000000038 chest Anatomy 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 230000001447 compensatory effect Effects 0.000 description 3
- 210000000918 epididymis Anatomy 0.000 description 3
- 238000012224 gene deletion Methods 0.000 description 3
- 230000000512 lipotoxic effect Effects 0.000 description 3
- 210000004115 mitral valve Anatomy 0.000 description 3
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 206010007559 Cardiac failure congestive Diseases 0.000 description 2
- 206010020772 Hypertension Diseases 0.000 description 2
- 206010022489 Insulin Resistance Diseases 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 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
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 2
- 206010071436 Systolic dysfunction Diseases 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000001746 atrial effect Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000002526 effect on cardiovascular system Effects 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 229960002725 isoflurane Drugs 0.000 description 2
- 210000005240 left ventricle Anatomy 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 238000007634 remodeling Methods 0.000 description 2
- 239000003001 serine protease inhibitor Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 230000002861 ventricular Effects 0.000 description 2
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- 208000031229 Cardiomyopathies Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010056370 Congestive cardiomyopathy Diseases 0.000 description 1
- 201000010046 Dilated cardiomyopathy Diseases 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000015779 HDL Lipoproteins Human genes 0.000 description 1
- 108010010234 HDL Lipoproteins Proteins 0.000 description 1
- 206010060378 Hyperinsulinaemia Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 208000017170 Lipid metabolism disease Diseases 0.000 description 1
- 241001417092 Macrouridae Species 0.000 description 1
- 208000001145 Metabolic Syndrome Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 229940122055 Serine protease inhibitor Drugs 0.000 description 1
- 101710102218 Serine protease inhibitor Proteins 0.000 description 1
- 102000008847 Serpin Human genes 0.000 description 1
- 108050000761 Serpin Proteins 0.000 description 1
- 235000009233 Stachytarpheta cayennensis Nutrition 0.000 description 1
- 206010042434 Sudden death Diseases 0.000 description 1
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000006793 arrhythmia Effects 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 210000002565 arteriole Anatomy 0.000 description 1
- 230000002567 autonomic effect Effects 0.000 description 1
- 230000008721 basement membrane thickening Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000004094 calcium homeostasis Effects 0.000 description 1
- 206010007625 cardiogenic shock Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000009091 contractile dysfunction Effects 0.000 description 1
- 230000008828 contractile function Effects 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 230000005713 exacerbation Effects 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 230000004129 fatty acid metabolism Effects 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 208000018914 glucose metabolism disease Diseases 0.000 description 1
- 235000009200 high fat diet Nutrition 0.000 description 1
- 230000003451 hyperinsulinaemic effect Effects 0.000 description 1
- 201000008980 hyperinsulinism Diseases 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 210000005246 left atrium Anatomy 0.000 description 1
- 235000019626 lipase activity Nutrition 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 230000004130 lipolysis Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 230000004065 mitochondrial dysfunction Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 208000031225 myocardial ischemia Diseases 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000001991 pathophysiological effect Effects 0.000 description 1
- 230000008289 pathophysiological mechanism Effects 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- DHRLEVQXOMLTIM-UHFFFAOYSA-N phosphoric acid;trioxomolybdenum Chemical compound O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.OP(O)(O)=O DHRLEVQXOMLTIM-UHFFFAOYSA-N 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- XOSXWYQMOYSSKB-LDKJGXKFSA-L water blue Chemical compound CC1=CC(/C(\C(C=C2)=CC=C2NC(C=C2)=CC=C2S([O-])(=O)=O)=C(\C=C2)/C=C/C\2=N\C(C=C2)=CC=C2S([O-])(=O)=O)=CC(S(O)(=O)=O)=C1N.[Na+].[Na+] XOSXWYQMOYSSKB-LDKJGXKFSA-L 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
- A61K38/57—Protease inhibitors from animals; from humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Veterinary Medicine (AREA)
- Immunology (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明公开了PEDF基因在治疗糖尿病性心肌损伤中的应用。本发明首次确定了PEDF基因和糖尿病性心肌损伤的关系，PEDF基因敲除可以导致体重和血糖升高、心肌细胞肥厚、心肌纤维化及心脏功能障碍。这表明PEDF基因在糖尿病性心肌损伤中的功能，主要体现在PEDF具有改善心肌细胞肥厚和纤维化、改善心脏功能的作用。针对PEDF基因的上述功能，PEDF可作为药物，用于预防、缓解或/和治疗糖尿病性心肌损伤；PEDF可作为药物靶点，用于筛选预防、缓解或/和治疗糖尿病性心肌损伤的药物；PEDF也可以作为基因治疗中的靶基因，用于设计和制备预防、缓解或/和治疗糖尿病性心肌损伤的药物和/或生物学制剂。
Description
技术领域
本发明涉及已知基因的功能与应用研究技术领域，具体地，涉及PEDF基因在治疗糖尿病性心肌损伤中的应用。
背景技术
心血管并发症是糖尿病患者病情加重和死亡的主要原因，约有 50～80%的糖尿病患者死于心血管疾病。心肌损害是最主要的糖尿病心血管并发症之一，糖尿病患者发生心力衰竭的风险是非糖尿病患者的2～4倍。自1972年美国心脏科医生Shirley Rubler等首次提出糖尿病心肌病（Diabetic cardiomyopathy，DCM）的概念以来，因糖尿病患病人数急剧增加可能造成心肌病发病率居高不下的现状，已引起学者和临床医生的极大关注。
糖尿病心肌病被认为是发生于糖尿病患者且独立于冠状动脉疾病及高血压的原发性心肌损害。目前研究认为，在无心肌缺血及高血压时，糖尿病患者的心脏结构及功能改变明显，表现出心肌细胞肥大、间质纤维化、PAS阳性物质浸润、冠状小动脉基底膜增厚、心肌内微血管病变等病理特征。临床表现为左室舒张和/或收缩功能障碍，或伴有心力衰竭、心源性休克，甚至猝死。糖尿病心肌病会造成患者心脏功能下降，并最终导致严重心律失常和心衰，显著降低患者的生活质量和寿命。因此，糖尿病心肌损伤的基础研究已成为“糖尿病心脏病理学”领域的前沿和热点，有效预防和治疗糖尿病心肌损伤也是临床医疗的重要目标之一。然而，直到现在，研究者对糖尿病心肌损伤病理生理机制的了解还不是很充分，临床上也缺乏糖尿病心肌损伤防治的有效策略和措施。已有研究显示，糖尿病心肌损害主要与高胰岛素血症、糖代谢障碍、脂毒性、线粒体功能障碍与氧化应激、钙稳态失衡、心肌微循环障碍、自主神经功能紊乱等因素有关。上述多重机制形成复杂的病理生理网络，共同推动糖尿病心肌损伤发生与发展。然而，越来越多的研究表明，以高游离脂肪酸为特征的脂毒性是糖尿病心肌损伤病理网络中的关键环节，能启动和诱导其他相关病理机制，进而引发心脏结构和功能的改变。
PEDF又称Serpin f1，是分泌性糖蛋白，属丝氨酸蛋白酶抑制剂超家族成员，具有明确的抑制血管新生、保护血管内皮和营养神经的作用与功能。不仅如此，PEDF还被证实是非常重要的脂代谢调节分子。临床研究数据显示，与正常人群相比，肥胖、代谢综合征和糖尿病患者外周血PEDF明显增高，而且其浓度还与甘油三酯呈正相关，与高密度脂蛋白呈负相关。在高脂饮食诱导肥胖并高脂血症小鼠模型上，我们发现PEDF能够双向调控甘油三酯脂酶的表达与活性。在肥胖早期，脂肪细胞PEDF通过增强甘油三酯脂酶活性而促进脂肪分解，使循环游离脂肪酸水平显著升高。此外，PEDF还能促进血浆游离脂肪酸在多组织器官异位沉积，诱导糖脂代谢紊乱和全身性胰岛素抵抗，这被认为是肥胖引发Ⅱ型糖尿病新的分子机制。但是，PEDF是否参与心肌细胞游离脂肪酸代谢与心脏脂毒性，是否是糖尿病心肌损伤病理过程中的关键分子，目前还不清楚。
发明内容
本发明的目的在于确定PEDF表达与糖尿病性心肌损伤的关系，提供PEDF在制备预防、缓解或/和治疗糖尿病性心肌损伤药物中的应用。
本发明的另一个目的是提供PEDF基因在作为筛选预防、缓解或/和治疗糖尿病性心肌损伤药物的药物靶标中的应用。
本发明的再一个目的是提供PEDF作为基因治疗中的靶基因，用于设计和制备预防、缓解或/和治疗糖尿病性心肌损伤药物和/或生物学制剂中的应用。
为了实现上述目的，本发明是通过以下技术方案予以实现的：
本发明通过研究首次确定了PEDF基因的表达和糖尿病性心肌损伤的关系。具体关系如下：1、PEDF基因缺失显著增加小鼠附睾周围脂肪堆积、体重和血糖；2、PEDF基因缺失显著增加小鼠心脏重量；3、PEDF基因缺失导致小鼠心肌肥厚；4、PEDF基因缺失导致小鼠心肌纤维化；5、PEDF基因缺失显著降低小鼠心脏功能。
由以上结果可知，PEDF敲除可导致小鼠体重和血糖显著升高，增加脂肪重量，增加心脏重量，并导致以心肌肥厚和纤维化为主要特征的心脏重塑，同时导致心脏功能下降。因此，PEDF具有改善小鼠糖尿病性心肌损伤的作用，为研究防治糖尿病性心肌损伤的新靶点和新策略提供了理论依据和临床基础。
所以，本发明要求保护PEDF在制备预防、缓解或/和治疗糖尿病性心肌损伤药物中的应用。所述的药物包括有效量的PEDF重组蛋白，以及药学上可接受的辅料。
本发明还要求保护PEDF基因在作为筛选预防、缓解或/和治疗糖尿病性心肌损伤药物的药物靶标中的应用。
本发明进一步要求保护PEDF基因作为基因治疗中的靶基因，用于设计和制备预防、缓解或/和治疗糖尿病性心肌损伤药物和/或生物学制剂中的应用。
所述的药物和/或生物制剂包含PEDF激活剂。
所述的糖尿病性心肌损伤为糖尿病引发的心肌肥大、心肌纤维化以及心脏功能障碍。
与现有技术相比，本发明具有如下有益效果：
本发明首次确定了PEDF与糖尿病性心肌损伤的关系，因此，PEDF可作为药物、药物靶点或基因治疗中的靶基因，应用于糖尿病性心肌损伤的预防、缓解或/和治疗，能够为糖尿病性心肌损伤的防治提供新的策略。此外，由于心肌损伤的发病机制并不明确，PEDF和糖尿病性心肌损伤关系的确定，将为进一步的研究糖尿病性心肌损伤的病因病机和相应的防治策略提供新的方向。
附图说明
图1是PEDF敲除小鼠及其同窝野生型小鼠56周龄的大体图片和体重统计图，结果显示PEDF敲除显著增加了小鼠体重；（***，p<0.001，和野生型组比较）。
图2是PEDF敲除小鼠及其同窝野生型小鼠56周龄的附睾脂肪垫重量和脂体比统计图，结果显示PEDF敲除显著增加了小鼠的脂肪堆积；（*，0.01<p<0.05，和野生型组比较）。
图3是PEDF敲除小鼠及其同窝野生型小鼠56周龄的血糖统计图，结果显示PEDF敲除显著增加了小鼠血糖；（**，0.001<p<0.01，和野生型组比较）。
图4是PEDF敲除小鼠及其同窝野生型小鼠56周龄的心脏大体图片和心脏重量、心胫比统计图，结果显示PEDF敲除显著增加了心脏重量；（*，0.01<p<0.05，和野生型组比较）。
图5是PEDF敲除小鼠及其同窝野生型小鼠56周龄的HE染色图片（上图标尺=2000μm；下图标尺=50μm）和心肌细胞大小倍数变化统计图，结果显示PEDF敲除明显导致小鼠心肌肥厚；（***，p<0.001，和野生型组比较）。
图6是PEDF敲除小鼠及其同窝野生型小鼠56周龄的Masson染色图片（标尺=50μm）和胶原面积比例统计图，结果显示PEDF敲除明显导致小鼠心肌纤维化；（***，p<0.001，和野生型组比较）。
图7是PEDF敲除小鼠及其同窝野生型小鼠56周龄的脉冲多普勒图片和E/A比值统计图，结果显示PEDF敲除显著增加了小鼠的E/A比值；（**，0.001<p<0.01，和野生型组比较）。
图8是PEDF敲除小鼠及其同窝野生型小鼠56周龄的M型超声图片和射血分数、缩短分数统计图，结果显示，PEDF敲除显著降低了小鼠的射血分数和缩短分数；（*，p<0.05，和野生型组比较）。
具体实施方式
下面结合说明书附图和具体实施例对本发明作出进一步地详细阐述，所述实施例只用于解释本发明，并非用于限定本发明的范围。下述实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，为可从商业途径得到的试剂和材料。
实验用动物及饲养：PEDF(-/-)小鼠为C57BL/6J背景，由美国OklahomaUniversity的马建兴教授课题组构建，并由中山大学中山医学院高国全教授课题组友情赠送。本发明所用实验动物均在中山大学疾病模式动物中心进行扩繁，扩繁方式为雄性PEDF(+/-)和雌性PEDF(+/-)交配扩繁。本发明实验动物选用雄性PEDF(-/-)小鼠及其同窝野生型雄性对照小鼠，各8～12只。于3周时剪鼠尾鉴定并分笼，一直观察到56周。期间自由饮水摄食，每12小时交替照明，温度21±2℃，湿度40%～70%。
小鼠终末组织取材：实验结束前禁食不禁水12小时。实验结束时，小鼠称重，剪尾测定血糖。腹腔注射1%戊巴比妥钠，80mg/kg，待小鼠麻醉后，将小鼠仰卧位固定于解剖板上，75%消毒酒精表面消毒。用手术剪剪开小鼠腹腔至胸腔，暴露心脏和脂肪。剪开右心耳，于左心室进针灌注PBS至血液流净。剪下心脏和附睾周围脂肪组织，于PBS中漂洗至无血渍后称重。剪下胫骨，测胫骨长度。
病理组织处理：取小鼠心脏，于PBS中漂洗至无血渍，4%多聚甲醛中固定过夜；PBS洗3次，每次30分钟；25%乙醇、50%乙醇、75%乙醇、90%乙醇、无水乙醇（Ⅰ）、无水乙醇（Ⅱ）梯度脱水，各30分钟；二甲苯（Ⅰ）、二甲苯（Ⅱ）各15分钟；石蜡（Ⅰ）、石蜡（Ⅱ）各60分钟；脱水后，心脏平放于石蜡中包埋；徕卡石蜡切片机切片，片厚5μm。
实施例1
小鼠体重、血糖和组织重量测定
实验结束前禁食不禁水12小时。实验结束时，小鼠称重，强生稳豪型血糖仪剪尾测血糖。腹腔注射1%戊巴比妥钠，80mg/kg，待小鼠麻醉后，进行大体拍照。随后将小鼠仰卧位固定于解剖板上，75%消毒酒精表面消毒。用手术剪剪开小鼠腹腔至胸腔，暴露心脏和脂肪。剪开右心耳，于左心室进针灌注PBS至血液流净。剪下心脏和附睾周围脂肪组织，于PBS中漂洗至无血渍后称重。对心脏进行拍照。剪下胫骨，测胫骨长度。计算脂体比和心胫比。
肥胖是导致胰岛素抵抗的主要因素，而胰岛素抵抗是糖尿病及脂代谢紊乱发生的基础。图1、图2结果表明，PEDF敲除可以显著增加小鼠体重和脂肪堆积，图3结果表明PEDF敲除可以显著升高小鼠血糖。
糖尿病性心肌损伤并发充血性心力衰竭时，会有心脏扩大等扩张型心肌病表现，图4结果表明，PEDF敲除明显导致心脏扩大。
实施例2
小鼠心肌肥厚测定
心脏石蜡切片HE染色，主要步骤为：60℃烘烤30分钟→二甲苯（Ⅰ）10分钟→二甲苯（Ⅱ）10分钟→无水乙醇（Ⅰ）3分钟→无水乙醇（Ⅱ）3分钟→95%乙醇（Ⅰ）1分钟→70%乙醇1分钟→蒸馏水2分钟→苏木素液5-10分钟→流水洗去苏木素1-3秒钟→1%盐酸酒精1-2秒钟→流水冲洗20分钟→蒸馏水过洗1-2秒钟→0.5%伊红2分钟→蒸馏水稍洗1-2秒钟→95%乙醇（Ⅱ）2-3秒钟→无水乙醇（Ⅲ）3-5秒钟→无水乙醇（Ⅳ）5-10秒钟→二甲苯（Ⅰ）2分钟→二甲苯（Ⅱ）2分钟→中性树胶封片观察→Image J软件分析细胞大小。
心肌肥厚是糖尿病性心肌损伤的主要病理特征，被认为是心肌细胞在长期高负荷状况下的一种代偿反应。心肌肥厚是导致心力衰竭的独立危险因素。图5结果显示，PEDF敲除明显导致心肌肥厚。
实施例3
小鼠心肌纤维化测定
心脏石蜡切片Masson染色，主要步骤为：60℃烘烤30分钟→二甲苯（Ⅰ）10分钟→二甲苯（Ⅱ）10分钟→无水乙醇（Ⅰ）3分钟→无水乙醇（Ⅱ）3分钟→95%乙醇（Ⅰ）1分钟→70%乙醇1分钟→蒸馏水2分钟→Weigert氏铁苏木素染5分钟→流水洗去苏木素1-3秒钟→1%盐酸酒精1-2秒钟→流水冲洗20分钟→蒸馏水过洗1-2秒钟→丽春红酸性品红液染5-10分钟→蒸馏水快速漂洗数秒→磷钼酸水溶液处理约3-5分钟→苯胺蓝液复染分钟→1%冰醋酸处理分钟→95%乙醇（Ⅱ）2-3秒钟→无水乙醇（Ⅲ）3-5秒钟→无水乙醇（Ⅳ）5-10秒钟→二甲苯（Ⅰ）2分钟→二甲苯（Ⅱ）2分钟→中性树胶封片观察→Image J软件分析胶原面积比例。
心肌纤维化是以心脏间质成纤维细胞过度增殖、胶原过度沉积及异常分布为特征的心脏间质重构，是糖尿病性心肌损伤的主要病理特征。图6结果表明，PEDF敲除明显导致心肌纤维化。
实施例4
小鼠心脏功能测定：利用Vevo 2100小动物超声仪检测小鼠心脏功能。小鼠异氟烷吸入麻醉，在麻醉状态下用脱毛膏对小鼠胸部脱毛，仰卧位置于恒温加热板上，持续性吸入异氟烷。小鼠四肢与心电图电极相连，用于检测心率并记录心电图。30MHz探头，于小鼠左胸骨部位探测。采用脉冲多普勒测定二尖瓣舒张早期峰值血流速度E和舒张晚期峰值血流速度A，计算E/A比值，以评价心脏舒张功能。短轴，行M型超声测量射血分数和缩短分数，以评价心脏收缩功能。
心功能障碍是糖尿病性心肌损伤的主要特征，在糖尿病性心肌损伤的早期，以舒张性功能障碍为主，在后期，以收缩性功能障碍为主。
二尖瓣舒张早期峰值血流速度E和舒张晚期峰值血流速度A的比值是评价心脏舒张功能的重要指标。在舒张功能障碍早期，由于舒张期左室压力增高引起E峰降低，左房容量和压力升高，而舒张晚期时左房代偿性收缩增强，使得A峰代偿性升高，导致E/A比值降低。在舒张功能障碍晚期，左房压力显著增加，舒张早期跨二尖瓣压力阶差增加，导致E峰幅度超正常化，使得E/A比值增加。图7结果表明，PEDF敲除显著降低了小鼠的心脏舒张功能。
射血分数和缩短分数是评价心脏收缩功能的重要指标，糖尿病性心肌损伤并发充血性心力衰竭时，会有收缩功能障碍的超声心动图表现，主要表现为射血分数和缩短分数下降。图8结果表明，PEDF敲除显著降低了小鼠的心脏收缩功能。
Claims (6)
1.PEDF在制备预防、缓解或/和治疗糖尿病性心肌损伤药物中的应用。
2.根据权利要求1所述的应用，其特征在于，所述的药物包括有效量的PEDF重组蛋白以及药学上可接受的辅料。
3.PEDF基因在作为筛选预防、缓解或/和治疗糖尿病性心肌损伤药物的药物靶标中的应用。
4.PEDF基因作为基因治疗中的靶基因用于设计和制备预防、缓解或/和治疗糖尿病性心肌损伤药物中的应用。
5.根据权利要求4所述的应用，其特征在于，所述的药物包含PEDF激活剂。
6.根据权利要求1至5任一项所述的应用，其特征在于，所述的糖尿病性心肌损伤为糖尿病引发的心肌肥大、心肌纤维化以及心脏功能障碍。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710819105.3A CN107693784B (zh) | 2017-09-12 | 2017-09-12 | Pedf基因在治疗糖尿病性心肌损伤中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710819105.3A CN107693784B (zh) | 2017-09-12 | 2017-09-12 | Pedf基因在治疗糖尿病性心肌损伤中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107693784A CN107693784A (zh) | 2018-02-16 |
| CN107693784B true CN107693784B (zh) | 2020-08-04 |
Family
ID=61172613
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710819105.3A Active CN107693784B (zh) | 2017-09-12 | 2017-09-12 | Pedf基因在治疗糖尿病性心肌损伤中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107693784B (zh) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112826823B (zh) * | 2021-01-11 | 2022-03-08 | 中国人民解放军军事科学院军事医学研究院 | 多西环素在制备治疗和/或预防和/或缓解和/或改善心肌病药物中的应用 |
| CN114452378B (zh) * | 2022-01-20 | 2023-06-16 | 中山大学 | Rdh10和/或其前药在制备治疗和/或预防糖尿病心肌损伤的药物中的应用 |
| CN114651785A (zh) * | 2022-03-28 | 2022-06-24 | 中山大学 | 一种基于PEDF/ApoE双基因敲除的非酒精性脂肪性肝炎小鼠模型构建方法及应用 |
| CN115089716B (zh) * | 2022-07-12 | 2023-04-21 | 齐齐哈尔医学院 | Bmp4作为作用靶点在制备治疗糖尿病心肌病药物中的应用 |
-
2017
- 2017-09-12 CN CN201710819105.3A patent/CN107693784B/zh active Active
Non-Patent Citations (2)
| Title |
|---|
| Pigment Epithelium-Derived Factor Mediates Autophagy and Apoptosis in Myocardial Hypoxia/Reoxygenation Injury;Hsuan-Fu Kuo,et al;《PLOS ONE》;20160524;1-14页 * |
| 糖尿病冠状动脉硬化性心脏病中心肌损伤与氧化应激的关系;孙洋;《中国循环杂志》;20141130;第29卷(第11期);952-954页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107693784A (zh) | 2018-02-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107693784B (zh) | Pedf基因在治疗糖尿病性心肌损伤中的应用 | |
| Tao et al. | Collagen XIV is important for growth and structural integrity of the myocardium | |
| Nemoto et al. | Escherichia coli LPS-induced LV dysfunction: role of toll-like receptor-4 in the adult heart | |
| Hwang et al. | Neutralization of interleukin-1β in the acute phase of myocardial infarction promotes the progression of left ventricular remodeling | |
| Horgan et al. | Murine models of diastolic dysfunction and heart failure with preserved ejection fraction | |
| Heather et al. | Isoproterenol induces in vivo functional and metabolic abnormalities; similar to those found in the infarcted rat heart | |
| JP7182162B2 (ja) | 心筋症、陳旧性心筋梗塞および慢性心不全の治療薬 | |
| KR101934328B1 (ko) | 아모디아퀸 및 항당뇨 약물을 유효성분으로 함유하는 당뇨병의 예방 또는 치료용 약학적 조성물 | |
| Lu et al. | Abrogation of lectin-like oxidized LDL receptor-1 attenuates acute myocardial ischemia-induced renal dysfunction by modulating systemic and local inflammation | |
| Li et al. | Syringin prevents cardiac hypertrophy induced by pressure overload through the attenuation of autophagy | |
| CN103948575A (zh) | 肉桂醛在制备促血管新生药物中的应用 | |
| US20240016769A1 (en) | Method for preparing compound or biological drug enhancing CNPase activity for treating heart diseases | |
| Zheng et al. | Krüppel-like transcription factor 11 (KLF11) overexpression inhibits cardiac hypertrophy and fibrosis in mice | |
| Tu et al. | Empagliflozin inhibits coronary microvascular dysfunction and reduces cardiac pericyte loss in db/db mice | |
| WO2021073249A1 (zh) | β-NMN在制备脓毒症器官损伤的治疗、预防药物中的应用 | |
| CN113207799B (zh) | 一种二型糖尿病小鼠快速心衰模型的构建方法 | |
| CN108704136A (zh) | 蛋白酶体抑制剂在制备预防、治疗心血管疾病的药物中的用途 | |
| CN108159399B (zh) | 一种凝血蛋白酶aPC在防治糖尿病心肌病药物中的应用 | |
| Tai et al. | CREG improves cardiac function by regulating cardiomyocytes’ autophagy in diabetic myocardial infarction rats. | |
| Li et al. | The effect of YiQiFuMai on ischemic heart failure by improve myocardial microcirculation and increase eNOS and VEGF expression | |
| CN114288301B (zh) | Dtq在制备急性心肌梗死治疗药物中的用途及相关产品 | |
| CN114632144B (zh) | Il-3在制备治疗或评估心脏损伤的产品中的应用 | |
| CN112245582B (zh) | Rab22a基因作为靶标在制备心肌梗死治疗产品中的用途及相关产品 | |
| Jun et al. | A4888 An Imaging-based Renal Denervation Strategy in Hypertensive Canines Using Integrated Ultrasound Catheter | |
| Lester | THE EFFECT OF METABOLIC FACTORS ON THE REGENERATION OF ZEBRAFISH TAIL FINS |
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 |