CN113876929A - 肽MOTS-c在制备治疗帕金森药物中的应用 - Google Patents
肽MOTS-c在制备治疗帕金森药物中的应用 Download PDFInfo
- Publication number
- CN113876929A CN113876929A CN202111389409.3A CN202111389409A CN113876929A CN 113876929 A CN113876929 A CN 113876929A CN 202111389409 A CN202111389409 A CN 202111389409A CN 113876929 A CN113876929 A CN 113876929A
- Authority
- CN
- China
- Prior art keywords
- mots
- peptide
- parkinson
- rat
- rats
- 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.)
- Granted
Links
- 101001028702 Homo sapiens Mitochondrial-derived peptide MOTS-c Proteins 0.000 title claims abstract description 40
- 102100037173 Mitochondrial-derived peptide MOTS-c Human genes 0.000 title claims abstract description 40
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 39
- 239000003814 drug Substances 0.000 title claims abstract description 23
- 229940079593 drug Drugs 0.000 title claims abstract description 13
- 208000018737 Parkinson disease Diseases 0.000 title claims description 24
- 238000002360 preparation method Methods 0.000 title abstract description 7
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 claims abstract description 46
- 229960003638 dopamine Drugs 0.000 claims abstract description 23
- 229940080817 rotenone Drugs 0.000 claims abstract description 19
- JUVIOZPCNVVQFO-UHFFFAOYSA-N rotenone Natural products O1C2=C3CC(C(C)=C)OC3=CC=C2C(=O)C2C1COC1=C2C=C(OC)C(OC)=C1 JUVIOZPCNVVQFO-UHFFFAOYSA-N 0.000 claims abstract description 19
- 210000005064 dopaminergic neuron Anatomy 0.000 claims abstract description 8
- 239000007928 intraperitoneal injection Substances 0.000 claims description 2
- 239000003223 protective agent Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 241000700159 Rattus Species 0.000 abstract description 56
- 210000001577 neostriatum Anatomy 0.000 abstract description 18
- 238000010171 animal model Methods 0.000 abstract description 6
- 210000004556 brain Anatomy 0.000 abstract description 6
- 208000024891 symptom Diseases 0.000 abstract description 6
- 230000006378 damage Effects 0.000 abstract description 5
- 210000001259 mesencephalon Anatomy 0.000 abstract description 5
- 230000003188 neurobehavioral effect Effects 0.000 abstract description 5
- 230000036542 oxidative stress Effects 0.000 abstract description 5
- 208000027418 Wounds and injury Diseases 0.000 abstract description 4
- 208000014674 injury Diseases 0.000 abstract description 4
- 229940035678 anti-parkinson drug Drugs 0.000 abstract 1
- 239000000939 antiparkinson agent Substances 0.000 abstract 1
- 210000003523 substantia nigra Anatomy 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 12
- 210000002569 neuron Anatomy 0.000 description 11
- 241001465754 Metazoa Species 0.000 description 10
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 8
- 230000033001 locomotion Effects 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 239000007924 injection Substances 0.000 description 7
- 238000002347 injection Methods 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 5
- 102000019197 Superoxide Dismutase Human genes 0.000 description 5
- 108010012715 Superoxide dismutase Proteins 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 229960003180 glutathione Drugs 0.000 description 5
- 229940118019 malondialdehyde Drugs 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000012071 phase Substances 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 230000008506 pathogenesis Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 229930040373 Paraformaldehyde Natural products 0.000 description 3
- 230000006399 behavior Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 210000003141 lower extremity Anatomy 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 210000003470 mitochondria Anatomy 0.000 description 3
- 230000004973 motor coordination Effects 0.000 description 3
- 230000004792 oxidative damage Effects 0.000 description 3
- 229920002866 paraformaldehyde Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- WYTHCOXVWRKRAH-LOKRTKBUSA-N CC[C@H](C)[C@H](NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)CNC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)CNC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](Cc1c[nH]c2ccccc12)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WYTHCOXVWRKRAH-LOKRTKBUSA-N 0.000 description 2
- 102000047174 Disks Large Homolog 4 Human genes 0.000 description 2
- 108700019745 Disks Large Homolog 4 Proteins 0.000 description 2
- 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 2
- 108020005196 Mitochondrial DNA Proteins 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000003542 behavioural effect Effects 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 101150069842 dlg4 gene Proteins 0.000 description 2
- 230000021824 exploration behavior Effects 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 238000011532 immunohistochemical staining Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 210000004558 lewy body Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 238000011552 rat model Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 239000000243 solution Substances 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
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010006100 Bradykinesia Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 206010017577 Gait disturbance Diseases 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 208000006083 Hypokinesia Diseases 0.000 description 1
- 206010022489 Insulin Resistance Diseases 0.000 description 1
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 1
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 1
- 208000002740 Muscle Rigidity Diseases 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 208000027089 Parkinsonian disease Diseases 0.000 description 1
- 206010034010 Parkinsonism Diseases 0.000 description 1
- 206010071390 Resting tremor Diseases 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 108091000117 Tyrosine 3-Monooxygenase Proteins 0.000 description 1
- 102000048218 Tyrosine 3-monooxygenases Human genes 0.000 description 1
- 208000028752 abnormal posture Diseases 0.000 description 1
- XBJFCYDKBDVADW-UHFFFAOYSA-N acetonitrile;formic acid Chemical compound CC#N.OC=O XBJFCYDKBDVADW-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004097 bone metabolism Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000007613 environmental 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
- DZGCGKFAPXFTNM-UHFFFAOYSA-N ethanol;hydron;chloride Chemical compound Cl.CCO DZGCGKFAPXFTNM-UHFFFAOYSA-N 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000005021 gait Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229960004502 levodopa Drugs 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000006609 metabolic stress Effects 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 231100000683 possible toxicity Toxicity 0.000 description 1
- 230000001242 postsynaptic effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000000946 synaptic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 238000001262 western blot Methods 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
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/10—Peptides having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Neurosurgery (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Psychology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Dermatology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明涉及一种肽MOTS‑c在制备治疗帕金森药物中的应用，本发明所述肽MOTS‑c能够降低鱼藤酮诱导的帕金森大鼠动物模型中脑氧化应激损伤水平，保护大鼠中脑黑质纹状体多巴胺能神经元，并减少多巴胺能神经元的丢失，显著提高纹状体的多巴胺水平，有效改善帕金森大鼠神经行为学症状。因此，肽MOTS‑c在制备抗帕金森药物中具有重要的应用意义。
Description
技术领域
本发明涉及医药技术领域，特别涉及一种肽MOTS-c在制备治疗帕金森药物中的应用。
背景技术
帕金森病(Parkinson’s disease，PD)是全球第二大常见的神经退行性疾病，其发病率随着年龄增长而增高，严重影响中老年人的身体健康及生活质量。PD的主要病理特征是中脑黑质多巴胺神经元的缺失和路易小体(Lewy body)形成，随着纹状体多巴胺的减少，出现静止性震颤、肌肉强直、运动迟缓、姿态步态异常等常见的临床表现，症状呈进行性发展，严重时患者生活不能自理。目前其病因尚未完全清楚，主要认为是遗传，环境和衰老等多种因素共同相互作用的结果,尤其是环境因素在散发性PD发病机制中具有重要作用。大量的流行病和小动物实验研究发现农药鱼藤酮与PD发病相关，鱼藤酮染毒大鼠可成功构建拟PD的动物模型。在已经公认的PD发病机制中，氧化应激是导致黑质纹状体多巴胺能神经元损伤的重要因素，在PD发病进程中发挥重要的作用。
目前PD主要以药物治疗为主，常用的多巴胺前体左旋多巴可以通过血脑屏障进入脑内，代谢为多巴胺后补充内源性多巴胺缺乏，以缓解PD的症状。但是，大多数患者在长期用药后，其疗效逐渐减退，且出现各种运动并发症(包括开关现象，易动症等)，反而进一步增加了治疗难度。尽管目前临床上开始采用多种联合用药的方法以延长药物使用时间，但是仍然未能取得良好的临床治疗效果。因此寻找治疗PD的新药是神经科学方向重要的研究方向。随着对PD发病机制认识的深入，从多环节入手，着重保护多巴胺能神经元也成为了新的治疗PD药物的研发思路之一。
线粒体衍生肽(mitochondrial derived peptides，MDPs)是线粒体DNA(Mitochondrial DNA，mtDNA)开放阅读框编码的具有生物活性的短肽，参与机体代谢稳态的调节，在人体内发挥广泛的生理作用。已有的研究指出，MOTS-c作为目前已知的三种内源性MDPs之一，其可参与调节线粒体生物医学功能，影响线粒体介导的代谢过程，是研究线粒体相关疾病重要的靶点。此外，肽MOTS-c可在代谢应激条件下保护细胞，还参与糖脂代谢等过程，并发挥调节胰岛素抵抗，改善脂质沉积，调节骨代谢等多种作用。不仅如此，由于多肽类药物具有高效能、高选择性、作用靶点广泛及潜在毒性低等优点，使得肽MOTS-c具有巨大的研究价值和应用前景。
然而，肽MOTS-c发挥作用的机制还未完全阐明，目前也没有相关肽MOTS-c可用于治疗帕金森病的相关报道。
发明内容
本发明的目的是提供肽MOTS-c在制备治疗帕金森药物中的应用，所述肽MOTS-c治疗由鱼藤酮诱导的帕金森效果显著，其可以降低大鼠中脑的氧化损伤水平，保护多巴胺神经元，显著减少纹状体多巴胺神经元的丢失，并提高纹状体多巴胺的含量，进一步改善大鼠神经行为学症状，为帕金森病的治疗药物提供了新的选择。
本发明的技术方案是：
肽MOTS-c在制备治疗帕金森药物中的应用。
所述帕金森为由鱼藤酮诱导的。
所述肽MOTS-c的氨基酸序列如SEQ.ID.NO.1所示。
所述药物为多巴胺能神经元的保护剂。
所述药物增加纹状体多巴胺的含量。
所述药物采用腹腔注射方式给药。
所述药物的用量为3-5mg/kg/日。
本发明用鱼藤酮诱导帕金森大鼠模型，鱼藤酮使用梯度给药，第一阶段注射剂量为2mg/kg，共给药3天。第二阶段注射剂量为1mg/kg，共给药7天。第三阶段注射剂量为0.5mg/kg，共给药20天。合计给药30天，建立鱼藤酮诱导大鼠帕金森动物模型。治疗组给予模型组腹腔注射MOTS-c，持续30天，剂量为3-5mg/kg。
申请人的动物实验证明，肽MOTS-c能显著改善鱼藤酮诱导的帕金森大鼠的神经行为症状。肽MOTS-c能降低帕金森大鼠中脑内的氧化损伤水平，进一步保护大鼠多巴胺神经元，减少纹状体多巴胺神经元的丢失，并显著提高纹状体内的多巴胺的含量。在行为学方面，大鼠在新环境下的自主探索行为和运动协调情况均得到了显著的改善。提示肽MOTS-c为治疗PD提供了一种新的思路。
附图说明
图1为大鼠中脑氧化损伤水平的检测结果，其中，
A为MDA(丙二醛)含量检测结果；图B为GSH(谷胱甘肽)含量的检测结果；图C为SOD(超氧化物歧化酶)活性的检测结果(注：*P<0.05，模型组与对照组比较；**P<0.01，模型组与对照组比较；#P<0.05，治疗组与模型组比较)
图2为大鼠纹状体HE染色结果，其中，箭头所指位置为细胞核。
图3为大鼠纹状体免疫组化的结果，其中，A为各组纹状体免疫组化染色(TH)结果；B为各组TH阳性面积的统计结果(注：*P<0.05，模型组与对照组比较；**P<0.01，模型组与对照组比较；#P<0.05，治疗组与模型组比较)。
图4大鼠纹状体蛋白免疫印迹的检测结果，其中，TH是多巴胺合成限速酶酪氨酸羟化酶，PSD95是突触后膜的标志物，SYP是突触前膜标志物。
图5为旷场实验结果，其中，A为大鼠运动总距离；B为大鼠运动轨迹的热图；C为大鼠后肢站立次数；D为静止不动时间(注：*P<0.05，模型组与对照组比较；**P<0.01，模型组与对照组比较；#P<0.05，治疗组与模型组比较)。
图6为转棒实验结果，图中，注：*P<0.05，模型组与对照组比较；**P<0.01，模型组与对照组比较；#P<0.05，治疗组与模型组比较。
具体实施方式
一、材料和试剂
1.动物
健康的SD雄性大鼠(5-7周龄)，体重220～270g之间，购于中国人民解放军陆军军医大学动物中心。
2.试剂
其他试剂均采用市售的分析纯产品。
3.主要仪器设备
二.实施例
实施例1动物处理
肽MOTS-c委托武汉百意欣生物科技有限公司合成，纯度为>95％，氨基酸序列如SEQ.ID.NO.1所示：
Met Arg Trp Gln Glu Met Gly Tyr Ile Phe Tyr Pro Arg Lys Leu Arg。
SD雄性大鼠44只，体重220～270g之间，饲养在室内温度为23±2℃的动物房，期间给大鼠提供自由饮水和进食。大鼠随机分为3组：对照组，模型组和治疗组。模型组(梯度剂量皮下注射)：第一阶段注射剂量为2mg/kg，共给药3天。第二阶段注射剂量为1mg/kg，共给药7天。第三阶段注射剂量为0.5mg/kg，共给药20天。合计给药30天，建立鱼藤酮诱导大鼠帕金森动物模型。治疗组：在模型组给药的同时，腹腔注射MOTS-c(3-5mg/kg)持续30天。处理结束后先检测大鼠行为学指标，包括旷场实验和转棒实验。行为学检测结束后麻醉处死大鼠，置于冰上迅速分离纹状体(一部分用多聚甲醛固定，一部分用液氮速冻)并收集其余脑组织。
实施例2实施各组大鼠神经行为学实验检测
1)旷场实验：旷场实验是评价实验动物在新环境中自主行为，探究行为以及紧张度的一种方法，以评价动物的活动度是否正常且均一。调整软件和系统参数后，将大鼠至于旷场反应箱内，让其自由活动10分钟，系统将记录大鼠的所有活动记录。实验结束后软件将记录分析其活动距离，活动轨迹和后肢站立次数等参数(每次实验前需要用酒精擦拭反应箱，以消除气味对动物的影响)。
2)转棒式疲劳仪实验：通过测量动物在滚轮上所停留的时间，用于评价动物平衡力，运动协调和中枢抑制等情况。实验开始前半小时将大鼠放到实验室适应环境。设置仪器转动参数，使得其以12rpm的速度匀速转动。实验动物依次放置于滚轮上，记录每只大鼠在转棒上保持平衡所能持续的时间，最长持续时间不超过3分钟。
实施例3大鼠纹状体多巴胺的含量(高分辨液质联用法)
将大鼠的纹状体称重并记录重量，精密加入1mL的甲醇(含0.1％甲酸)，漩涡震荡1min，匀浆3min。高速离心14000rpm，10min，取上清进样分析。色谱Waters T3(150×2.1mm，3μm)。流速0.3mL/min，水相0.1％甲酸水溶液，有机相0.1％甲酸乙腈，洗针液甲醇，柱温箱温度35℃。
实施例4免疫组织化学染色
纹状体组织用4％多聚甲醛固定后，常规脱水，石蜡包埋，切片，片厚5μm。组织切片经脱蜡水化，抗原修复，封闭内源性过氧化氢酶，抗体杂交(TH抗体)，DAB显色，复染，脱水，透明和封片后镜检。进一步用组织切片数字扫描仪扫描图像，应用赛维尔图像分析软件自动读取组织测量区域，首先进行阳性等级划分：阴性无着色，计为0分；弱阳淡黄色，计为1分，中阳性棕黄色，记为2分；强阳性棕褐色，计为3分，最终分析计算测量区域内阳性面积比，反应阳性面积的多少。
实施例5HE染色
纹状体组织用4％多聚甲醛固定后，常规脱水，石蜡包埋，切片，片厚5μm。切片经过脱蜡水化后按照HE试剂盒说明书进行苏木素-伊红染色，苏木素染核，盐酸乙醇分化，伊红染细胞质，透明化后封片。在光学显微镜下观察纹状体结构及损伤改变，并进行拍照。
实施例6免疫印迹法
纹状体组织提取总蛋白；BCA法进行蛋白浓度测定；然后进行SDS-聚丙烯酰胺凝胶电泳，转膜，封闭，一抗孵育过夜(4度)，对应种属二抗孵育(室温)，化学发光液进行显影。拍照后用Image J软件进行灰度值分析。
实施例7大鼠脑组织内GSH，MDA和SOD的含量测定
GSH，MDA和SOD检测试剂盒均购自上海碧云天生物科技有限公司，操作步骤均按照试剂盒说明书进行。
三.结果
1、肽MOTS-c可缓解鱼藤酮帕金森大鼠中脑氧化应激损伤
结果显示，治疗组大鼠大脑内MDA含量有所下降，同时GSH含量和SOD活性也有所提升，表明肽MOTS-c对鱼藤酮帕金森大鼠中脑内的氧化应激损伤水平有明显缓解作用(见图1)。
2、肽MOTS-c减轻鱼藤酮帕金森大鼠纹状体多巴胺神经元损伤
HE染色结果显示(见图2)，治疗组纹状体组织形态基本规则，细胞形态虽有回缩但接近于正常，核固缩现象明显好转。表明肽MOTS-c对鱼藤酮大鼠纹状体神经元有显著保护作用。
3、肽MOTS-c可减少帕金森大鼠纹状体多巴胺神经元丢失
如图所示(图3，图4)，模型组大鼠纹状体TH阳性着色减少明显，同时纹状体TH蛋白表达水平下降，提示帕金森大鼠纹状体多巴胺能神经元有显著丢失，这也是PD的重要标志。而治疗组大鼠纹状体的TH阳性细胞数量和TH蛋白表达水平明显提高。此外，治疗组纹状神经元突触标志物PSD95和SYP的表达水平相较模型组显著增高，上述结果提示MOTS-c对纹状体多巴胺神经元有明显的保护作用。
4、肽MOTS-c可增加帕金森大鼠纹状体多巴胺含量
表1结果显示，模型组纹状体DA含量显著降低，进一步说明模型复制成功；肽MOTS-c能显著提高纹状体多巴胺水平的作用。
表1：3组大鼠纹状体DA含量的测定结果
(注：*P<0.05，模型组与对照组比较；**P<0.01，模型组与对照组比较；#P<0.05，治疗组与模型组比较)
5、肽MOTS-c可改善鱼藤酮帕金森大鼠探究能力
旷场实验结果如图5所示，模型组大鼠后肢站立次数减少，运动距离缩短，休息时间增加，表明模型组动物在新环境中的自主探究行为能力显著下降，也提示帕金森模型复制成功。而治疗组的上述指标都得到了一定程度的好转，提示肽MOTS-c给药可以改善鱼藤酮致帕金森大鼠新环境下自主探究行为的能力。
6、肽MOTS-c可改善鱼藤酮帕金森大鼠运动协调能力
模型组大鼠运动协调障碍明显，且四肢活动欠佳，其在转棒上活动的时间显著减短，甚至个别大鼠活动极度不协调，很难在转棒上持续活动。而治疗组大鼠运动协调能力显著改善，在转棒上活动的时间明显延长。提示肽MOTS-c对缓解帕金森大鼠运动协调能力有积极的作用(见图6)。
综上，本发明建立鱼藤酮诱导帕金森大鼠模型，同时给予肽MOTS-c进行干预。实验结果显示肽MOTS-c能够缓解帕金森大鼠大脑的氧化应激损伤水平，保护多巴胺神经元，减少多巴胺神经元的丢失，且提高纹状体多巴胺的含量，改善帕金森大鼠的神经行为学症状。因此，肽MOTS-c对帕金森大鼠有明显的保护作用。肽MOTS-c能够应用于制备治疗帕金森病的药物。
序列表
<110> 中国人民解放军陆军军医大学
<120> 肽MOTS-c在制备治疗帕金森药物中的应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 16
<212> PRT
<213> 人工合成(Artificial synthesis)
<400> 1
Met Arg Trp Gln Glu Met Gly Tyr Ile Phe Tyr Pro Arg Lys Leu Arg
1 5 10 15
Claims (7)
1.肽MOTS-c在制备治疗帕金森药物中的应用。
2.根据权利要求1所述的应用，其特征在于：所述帕金森为由鱼藤酮诱导的。
3.根据权利要求1所述的应用，其特征在于：所述肽MOTS-c的氨基酸序列如SEQ.ID.NO.1所示。
4.根据权利要求1所述的应用，其特征在于：所述药物为多巴胺能神经元的保护剂。
5.根据权利要求1所述的应用，其特征在于：所述药物可增加纹状体多巴胺的含量。
6.根据权利要求1所述的应用，其特征在于：所述药物采用腹腔注射的方式给药。
7.根据权利要求1所述的应用，其特征在于：所述药物的用量为3-5mg/kg/日。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111389409.3A CN113876929B (zh) | 2021-11-22 | 2021-11-22 | 肽MOTS-c在制备治疗帕金森药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111389409.3A CN113876929B (zh) | 2021-11-22 | 2021-11-22 | 肽MOTS-c在制备治疗帕金森药物中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113876929A true CN113876929A (zh) | 2022-01-04 |
| CN113876929B CN113876929B (zh) | 2024-02-23 |
Family
ID=79015472
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111389409.3A Active CN113876929B (zh) | 2021-11-22 | 2021-11-22 | 肽MOTS-c在制备治疗帕金森药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113876929B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115486415A (zh) * | 2022-08-11 | 2022-12-20 | 中国农业大学 | 蜂帕金森模型的建立方法及应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170049853A1 (en) * | 2013-03-15 | 2017-02-23 | The Regents Of The University Of California | Mitochondrial-derived peptide mots3 regulates metabolism and cell survival |
| CN110072883A (zh) * | 2016-09-28 | 2019-07-30 | 科巴公司 | 治疗性mots-c相关的肽 |
| CN110818804A (zh) * | 2019-10-21 | 2020-02-21 | 兰州大学 | 脑靶向肽及其在制备增强记忆药物中的应用 |
| US20210030835A1 (en) * | 2018-03-27 | 2021-02-04 | Cohbar, Inc. | Peptide-containing formulations |
-
2021
- 2021-11-22 CN CN202111389409.3A patent/CN113876929B/zh active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20170049853A1 (en) * | 2013-03-15 | 2017-02-23 | The Regents Of The University Of California | Mitochondrial-derived peptide mots3 regulates metabolism and cell survival |
| CN110072883A (zh) * | 2016-09-28 | 2019-07-30 | 科巴公司 | 治疗性mots-c相关的肽 |
| US20200181197A1 (en) * | 2016-09-28 | 2020-06-11 | Cohbar, Inc. | Therapeutic peptides |
| US20210030835A1 (en) * | 2018-03-27 | 2021-02-04 | Cohbar, Inc. | Peptide-containing formulations |
| CN110818804A (zh) * | 2019-10-21 | 2020-02-21 | 兰州大学 | 脑靶向肽及其在制备增强记忆药物中的应用 |
Non-Patent Citations (3)
| Title |
|---|
| AMICA C MÜLLER-NEDEBOCK ET AL.: "The unresolved role of mitochondrial DNA in Parkinson\'s disease: An overview of published studies, their limitations, and future prospects", NEUROCHEM INT, vol. 129, 31 October 2019 (2019-10-31), pages 1 - 34 * |
| SU-JEONG KIM ET AL.: "Mitochondrial peptides modulate mitochondrial function during cellular senescence", AGING (ALBANY NY), vol. 10, no. 6, 10 June 2018 (2018-06-10), pages 1239 - 1256 * |
| 庞笑丽 等: "线粒体衍生肽MOTS-c生物学特性及病理生理功能", 生命的化学, vol. 41, no. 9, 18 September 2021 (2021-09-18), pages 1927 - 1934 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115486415A (zh) * | 2022-08-11 | 2022-12-20 | 中国农业大学 | 蜂帕金森模型的建立方法及应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113876929B (zh) | 2024-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | GIP has neuroprotective effects in Alzheimer and Parkinson’s disease models | |
| Lima et al. | Neuroprotective activities of Spirulina platensis in the 6-OHDA model of Parkinson’s disease are related to its anti-inflammatory effects | |
| Li et al. | Puerarin attenuates neuronal degeneration in the substantia nigra of 6-OHDA-lesioned rats through regulating BDNF expression and activating the Nrf2/ARE signaling pathway | |
| Gao et al. | Fucoidan, a sulfated polysaccharide from brown algae, improves cognitive impairment induced by infusion of Aβ peptide in rats | |
| Tang et al. | Neuroprotective effects of tanshinone IIA and/or tetramethylpyrazine in cerebral ischemic injury in vivo and in vitro | |
| Cai et al. | A GLP-1/GIP Dual Receptor Agonist DA4-JC Effectively Attenuates Cognitive Impairment and Pathology in the APP/PS1/Tau Model of Alzheimer’s Disease 1 | |
| EP3505530B1 (en) | Scorpion venom heat-resistant synthesized peptide and uses thereof | |
| Jiang et al. | Inhibitory activities of kukoamines A and B from Lycii Cortex on amyloid aggregation related to Alzheimer’s disease and type 2 diabetes | |
| CN113876929A (zh) | 肽MOTS-c在制备治疗帕金森药物中的应用 | |
| Liu et al. | Melatonin alters amino acid metabolism and inflammatory responses in colitis mice | |
| Li et al. | Coeloglossum viride var. bracteatum extract attenuates Aβ-induced toxicity by inhibiting RIP1–driven inflammation and necroptosis | |
| Han et al. | Changes in insulin-signaling transduction pathway underlie learning/memory deficits in an Alzheimer’s disease rat model | |
| CN108473533B (zh) | 新型抗微生物肽及其用途 | |
| CN106456606A (zh) | 吲哚基及吲哚啉基异羟肟酸于治疗神经退化病症或认知缺乏的用途 | |
| CN110041408B (zh) | 一种小分子多肽及其在制备防治帕金森综合症药物中的应用 | |
| Mendieta et al. | The C-terminal domain of the heavy chain of tetanus toxin given by intramuscular injection causes neuroprotection and improves the motor behavior in rats treated with 6-hydroxydopamine | |
| He et al. | HSPA1A ameliorated spinal cord injury in rats by inhibiting apoptosis to exert neuroprotective effects | |
| CN113491763B (zh) | 眼镜蛇神经毒素及其制剂在制备预防和/或治疗帕金森病的药物中的应用 | |
| Yang et al. | Mesencephalic astrocyte-derived neurotrophic factor: a treatment option for Parkinson’s disease | |
| Yao et al. | Pretreatment with intravenous FGF-13 reduces infarct volume and ameliorates neurological deficits following focal cerebral ischemia in rats | |
| Dabravolski | Mitochondria-derived peptides in healthy ageing and therapy of age-related diseases | |
| CN111870683A (zh) | 一种抗菌肽在制备药物或化妆品中的用途 | |
| Huang et al. | Dendrobium nobile Lindl. alkaloid decreases Tau hyperphosphorylation via regulating PI3K/Akt/GSK-3β pathway in vitro and in vivo | |
| CN111358784B (zh) | 己糖激酶抑制剂在制备预防和/或治疗阿尔茨海默病的药物中的用途 | |
| Lin et al. | Mechanisms of Cong Rong Shu Jing Compound Effects on Endoplasmic Reticulum Stress in a Rat Model of Parkinson’s Disease |
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 |