CN110227146A - Metrnl蛋白或基因在防治认知障碍方面的应用 - Google Patents
Metrnl蛋白或基因在防治认知障碍方面的应用 Download PDFInfo
- Publication number
- CN110227146A CN110227146A CN201810183174.4A CN201810183174A CN110227146A CN 110227146 A CN110227146 A CN 110227146A CN 201810183174 A CN201810183174 A CN 201810183174A CN 110227146 A CN110227146 A CN 110227146A
- Authority
- CN
- China
- Prior art keywords
- metrnl
- gene
- albumen
- knock
- present
- 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.)
- Pending
Links
- 101150115803 metrnl gene Proteins 0.000 title claims abstract description 77
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 28
- 208000010877 cognitive disease Diseases 0.000 title abstract description 10
- 230000019771 cognition Effects 0.000 claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 8
- 229940079593 drug Drugs 0.000 claims abstract description 8
- 230000004064 dysfunction Effects 0.000 claims abstract description 6
- 230000002265 prevention Effects 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims description 11
- 235000013305 food Nutrition 0.000 claims description 5
- 230000036541 health Effects 0.000 claims description 5
- 239000000556 agonist Substances 0.000 claims description 3
- 239000003381 stabilizer Substances 0.000 claims description 3
- 238000011813 knockout mouse model Methods 0.000 abstract description 19
- 230000013016 learning Effects 0.000 abstract description 14
- 241000699670 Mus sp. Species 0.000 abstract description 13
- 230000006386 memory function Effects 0.000 abstract description 10
- 230000014509 gene expression Effects 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 7
- 210000001320 hippocampus Anatomy 0.000 abstract description 7
- 206010012289 Dementia Diseases 0.000 abstract description 6
- 230000002490 cerebral effect Effects 0.000 abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 6
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- 238000012347 Morris Water Maze Methods 0.000 abstract description 5
- 201000010099 disease Diseases 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 5
- 230000006698 induction Effects 0.000 abstract description 4
- 230000009885 systemic effect Effects 0.000 abstract description 4
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 abstract description 3
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 abstract description 3
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 abstract description 3
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 26
- 241001465754 Metazoa Species 0.000 description 23
- 238000002474 experimental method Methods 0.000 description 12
- 238000012360 testing method Methods 0.000 description 11
- 210000004556 brain Anatomy 0.000 description 10
- 239000000047 product Substances 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 8
- 244000144993 groups of animals Species 0.000 description 7
- 108700019745 Disks Large Homolog 4 Proteins 0.000 description 6
- 102000047174 Disks Large Homolog 4 Human genes 0.000 description 6
- 102000053171 Glial Fibrillary Acidic Human genes 0.000 description 6
- 101710193519 Glial fibrillary acidic protein Proteins 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 230000003920 cognitive function Effects 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 210000005046 glial fibrillary acidic protein Anatomy 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 230000000975 bioactive effect Effects 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 239000008194 pharmaceutical composition Substances 0.000 description 5
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 4
- 101150069842 dlg4 gene Proteins 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 210000001130 astrocyte Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 102000003952 Caspase 3 Human genes 0.000 description 2
- 108090000397 Caspase 3 Proteins 0.000 description 2
- 102000006587 Glutathione peroxidase Human genes 0.000 description 2
- 108700016172 Glutathione peroxidases Proteins 0.000 description 2
- 102100039669 Meteorin-like protein Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 102000004874 Synaptophysin Human genes 0.000 description 2
- 108090001076 Synaptophysin Proteins 0.000 description 2
- 108090000704 Tubulin Proteins 0.000 description 2
- 102000004243 Tubulin Human genes 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- 239000002552 dosage form Substances 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
- 150000002632 lipids Chemical class 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- -1 upper adjustment Substances 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 101001034286 Homo sapiens Meteorin-like protein Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 208000026139 Memory disease Diseases 0.000 description 1
- 101710146512 Meteorin-like protein Proteins 0.000 description 1
- 101150100800 Metrn gene Proteins 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 102000007072 Nerve Growth Factors Human genes 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000005775 apoptotic pathway Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000005754 cellular signaling Effects 0.000 description 1
- 210000004720 cerebrum Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000003792 cranial nerve Anatomy 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 210000003027 ear inner Anatomy 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000008451 emotion Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 208000016354 hearing loss disease Diseases 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 238000003754 machining Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 239000003900 neurotrophic factor Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000027272 reproductive process Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 230000000946 synaptic effect Effects 0.000 description 1
- 230000003956 synaptic plasticity Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/185—Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- 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
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Neurology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Neurosurgery (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Psychology (AREA)
- Hospice & Palliative Care (AREA)
- Psychiatry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及Metrnl蛋白或基因在防治认知障碍方面的应用。本发明采用Metrnl全身性敲除小鼠及相应的野生型对照小鼠，制备D‑半乳糖诱导的痴呆模型，运用Morris水迷宫实验对学习与记忆功能进行检测，结果显示：与野生型对照小鼠比较，Metrnl敲除小鼠的学习记忆功能下降，大脑海马区的脑源性神经营养因子等蛋白表达降低，表明Metrnl可用于认知功能障碍类疾病的防治。本发明为防治认知障碍提供了一种新方法，且由于Metrnl本身是一种人体内源性物质，其对人体可能的副作用很小，因此作为潜在药物的安全性很高。
Description
技术领域
本发明涉及医药技术领域，具体地说，是Metrnl蛋白或基因在防治认知障碍方面的应用。
背景技术
认知是机体认识和获取知识的智能加工过程，涉及学习、记忆、语言、思维、精神、情感等行为。认知障碍指与上述学习记忆以及思维判断有关的大脑高级智能加工过程出现异常，从而引起学习、记忆障碍。认知是大脑的正常功能，任何引起大脑功能和结构异常的因素均可导致认知障碍。随着世界向老龄社会进入步伐的不断加快，认知障碍、老年痴呆(阿尔茨海默病)等疾病已经成为威胁人类晚年健康，降低老年人生活质量的重要危险因素，给社会及家庭带来了沉重的经济负担和精神压力。因此，开发改善认知功能障碍的药物成为医疗工作者的研究重点。
Metrnl又称Subfatin、Cometin、Interleukin39，是近年来利用限食模型找到的一个新的脂肪因子，其在皮下脂肪组织、肠道上皮组织等部位高表达，与Metrn是同源蛋白，但表达分布却大不相同。目前，关于Metrnl功能的研究较少。国际专利公布号WO2010009732A1和中国专利公布号CN102164611A公开了Metrnl在减轻内耳听力损害方面的应用。中国专利公布号CN103536903A、CN103536904A和国际专利公布号WO2015062167A8公开了Metrnl在降血脂、降血糖中的应用。中国专利公布号CN104808004A公开了Metrnl在诊断结肠癌中的应用。但是，关于Metrnl在防治认知障碍类疾病方面的应用目前尚未见报道。
发明内容
本发明的目的是针对现有技术中的不足，提供Metrnl蛋白或基因的新用途。
本发明的第一方面，提供Metrnl蛋白或基因或它们的增效剂在制备防治认知功能障碍的药物中的应用。
本发明的第二方面，提供Metrnl蛋白或基因或它们的增效剂在制备改善学习记忆能力的药物中的应用。
本发明的第三方面，提供Metrnl蛋白或基因或它们的增效剂在制备改善认知功能障碍的食品和保健品中的应用。
本发明的第四方面，提供Metrnl蛋白或基因或它们的增效剂在制备改善学习记忆能力的食品和保健品中的应用。
作为本发明的一种优选实施方式，所述的增效剂选自激动剂、上调剂或稳定剂等。
本发明优点在于：
本发明采用Metrnl全身性敲除小鼠及相应的野生型对照小鼠，制备D-半乳糖诱导的痴呆模型，运用Morris水迷宫实验对学习与记忆功能进行检测，结果显示：与野生型对照小鼠比较，Metrnl敲除小鼠的学习记忆功能下降，大脑海马区的脑源性神经营养因子等蛋白表达降低，表明Metrnl可用于认知功能障碍类疾病的防治。本发明为防治认知障碍提供了一种新方法，且由于Metrnl本身是一种人体内源性物质，其对人体可能的副作用很小，因此作为潜在药物的安全性很高。
附图说明
附图1为D-gal模型学习记忆功能下降。生理盐水组，n＝10；D-gal组，n＝12。
附图2为D-gal模型脑组织的细胞凋亡和脂质过氧化损伤增加。生理盐水组，n＝10；D-gal组，n＝12。
附图3为雌性Metrnl敲除小鼠制备D-gal模型实验结果。在雌性动物D-gal模型上，与野生型小鼠比较，Metrnl敲除小鼠的学习记忆功能下降；WT组，n＝12；KO组，n＝16。
附图4为雄性Metrnl敲除小鼠制备D-gal模型实验结果。在雄性动物D-gal模型上，与野生型小鼠比较，Metrnl敲除小鼠的学习功能下降；WT组，n＝17；KO组，n＝17。
附图5为雄性Metrnl敲除小鼠制备D-gal模型大脑海马区Western-blot实验结果。左图：在D-gal雄性动物模型上，与野生型小鼠比较，Metrnl敲除小鼠的大脑海马区BDNF、GFAP和PSD95蛋白表达下降；右图：在不造模条件下，这些蛋白表达在KO组与WT组之间没有明显差别。
具体实施方式
下面结合具体实施方式，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。此外应理解，在阅读了本发明记载的内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
Metrnl蛋白及基因
本文中，术语“Metrnl蛋白”等同于“Metrnl多肽”，二者可互换使用。
本文中，所用的Metrnl蛋白可以是天然存在的，比如其可被分离纯化自哺乳动物。
此外，所述Metrnl蛋白也可以是人工制备的，比如根据常规基因工程技术来制备得到。任何合适的Metrnl蛋白均可适用于本发明。所述Metrnl蛋白包括全长的Metrnl蛋白或其生物活性片段。作为一种具体实施方式，所述Metrnl蛋白可以是一种人Metrnl蛋白，大小为30kDa左右，氨基酸序列见NP_001004431.1。
经过一个或多个氨基酸残基的取代、缺失或添加而形成的Metrnl蛋白的氨基酸序列也包括在本发明中。Metrnl蛋白或其生物活性片段包括一部分保守氨基酸的替代序列，所述氨基酸替代的序列并不影响其活性或保留了其部分活性。适当替换氨基酸是本领域内的公知技术，所述技术可以很容易地被实施并且确保不改变已知分子的生物活性。这些技术使本领域人员认识到，一般来说，在一种多肽的非必需氨基酸区域改变单个氨基酸并不会改变生物活性。
任何一种Metrnl蛋白的生物活性片段均可以应用到本发明中。在这里，Metrnl蛋白的生物活性片段的含义是指一种多肽，其仍然能保持全长的Metrnl蛋白的全部或部分功能。通常情况下，所述的生物活性片段至少保持50％、60％至99％或者100％的全长Metrnl蛋白的活性。
本发明也可以采用经修饰或改良的全部或部分氨基酸的Metrnl蛋白，比如，可以为了促进半衰期、有效性、代谢和/或蛋白的效力而加以修饰或改良的Metrnl蛋白。所述经过修饰或改良的Metrnl蛋白可以是一种Metrnl蛋白的共轭物，或其可被取代的或人工的氨基酸。所述经过修饰或改良的Metrnl蛋白或基因可以与天然Metrnl蛋白或基因有一定的不同点，但也具有本发明所述的功能，且不会带来其它不良反应或毒性。也就是说，任何不影响Metrnl蛋白的生物活性或者说是基因的生物学功能的变化形式都可用于本发明中。
Metrnl增效剂及其用途
所述的“Metrnl增效剂”包括了激动剂、上调剂、稳定剂等，是指任何可提高Metrnl的活性、提高Metrnl的稳定性、上调Metrnl的表达、增加Metrnl有效作用时间的物质，这些物质均可用于本发明。它们可以是化合物、化学小分子、生物分子等。所述的生物分子可以是核酸水平(包括DNA、RNA)的，蛋白水平的，也可以是上调Metrnl表达的病毒产品等。
药物组合物
依据本发明的方法，Metrnl蛋白或基因或它们的增效剂可以制备防治认知功能障碍类疾病的药物组合物，所述的药物组合物可由药物活性成分Metrnl蛋白或基因或它们的增效剂和药学上可接受的载体组成。
所述的药物组合物可采用医学领域常规的方法，将Metrnl蛋白或基因或它们的增效剂作为活性成分，与药学上可接受的辅料制成各种剂型。当用于口服时，可将其制备成常规的固体制剂如片剂、粉剂或胶囊剂等；用于注射时，可将其制备成注射剂。在各种制剂中，活性成分的重量含量为0.01％～99.9％。
所述的药物组合物可以用于治疗认知功能障碍类疾病。可以按剂型通过口服、腹腔注射、皮下注射、静脉注射、肌肉注射、粘膜用药等途径应用于需要治疗的个体，所述个体可以是人或动物。
实施例D-半乳糖诱导的痴呆模型的制备及Metrnl蛋白参与大脑认知功能的调控
一、实验方法
1.基因工程实验动物的培育。Metrnl全身性敲除小鼠采用Cre-LoxP条件性基因敲除系统(Diabetes 2015；64:4011-4022)，经交配繁殖而得。交配繁殖过程需要三种小鼠，其来源为：MetrnlloxP/loxP为我们独家拥有(Diabetes 2015；64:4011-4022)，Ella-Cre小鼠购自上海南方模式生物科技发展有限公司，C57小鼠购自上海西普尔-必凯实验动物有限公司。交配繁殖后可同时获得Metrnl全身性敲除小鼠(knock out,KO)及野生型正常对照小鼠(wild type,WT)。
2.D-半乳糖(D-galactose,D-gal)诱导的痴呆模型的制备。该模型主要原理是过量的D-gal在体内蓄积，在代谢的过程中可以产生大量的活性氧，对机体造成氧化应激损伤。D(+)galactose购自生工生物工程(上海)股份有限公司，产品编号A600215-0025。造模时，选择8～12周龄的小鼠，常选择雌性小鼠，造模组连续腹腔注射D-gal生理盐水溶液60天，剂量为100～200mg/kg；溶剂对照组连续腹腔注射10ml/kg体积的生理盐水溶液60天。
3.Morris水迷宫(Morris water maze,MWM)学习记忆功能检测。该实验是开展学习和记忆研究的首选经典实验。实验采集分析系统(型号XR-XM101，购自上海欣软信息科技有限公司)位于一个独立的实验室内，以避免外界干扰。在黑色水盆(直径120cm、高度35cm)内注水，水温保持在22±1℃，用无毒染料将水染成白色；水池人为分成四个象限，将一个直径10cm的透明平台固定在其中一个象限的中心，保持在水面以下1cm，水池周围用窗帘遮挡，并悬挂不同颜色和形状的塑料板构成方向标志物，水池上方设置视频跟踪监测系统，连接在计算机上。实验连续进行，共计6天：第1天到第5天是空间采集实验(acquisitiontrial)，第6天是探测试验(probe trial)。在前5天，动物在盆壁四个固定位置面对盆壁投放入水，每天按不同的顺序进行。在每次试验中，给动物60s时间找到隐藏在水下的平台，如果无法在60s内找到平台，则引导动物到平台并停留15s。实验顺序按照动物序号依次进行。由视频跟踪监测系统自动记录动物到达平台的潜伏期、游泳距离等指标。在第6天，撤去平台，将动物在平台所在象限的对侧盆壁最远处投放入水，由计算机自动记录动物穿越平台区域次数和在平台所在象限的停留时间。之后对上述参数进行分析。
4.分子指标检测。采用试剂盒测定的分子指标包括Caspase3、丙二醛(MDA)、总SOD、谷胱甘肽过氧化物酶(GSH-px)，试剂盒产品编号分别为C1116、S0131、S0101、S0056，均购自碧云天生物技术有限公司；采用Western-blot方法测定的分子指标包括BDNF(brainderived neurotrophic factor)、PSD95(postsynaptic density protein-95)、Synaptophysin、GFAP(glial fibrillary acidic protein)、Tubulin，其中：BDNF、Synaptophysin、GFAP抗体产品编号分别为ab205067、ab14692、ab7260，购自Abcam公司；PSD95抗体产品编号为2507S，购自Cell Signaling公司；Tubulin抗体产品编号为AT819，购自碧云天生物技术有限公司。
5.统计学分析。实验数据除Morris水迷宫空间采集实验采用平均值±标准误(Mean±SEM)之外，其余均采用平均值±标准差(Mean±SD)，两组之间比较时，计量资料实验数据采用t检验，计数资料实验数据采用Mann-Whitney U秩和检验，p＜0.05时认为两组有统计学差异(图中*代表p＜0.05；***代表p＜0.001)。
二、实验结果
1.正常野生型小鼠制备D-gal模型实验结果。见图1，从前五天的空间采集实验中可见，在第4天和第5天两组动物到达平台的潜伏期有统计学差异，给予D-gal的模型组动物寻找平台的时间明显长于对照组给予生理盐水的动物，说明模型组动物的学习能力较差；在第6天的探测试验中可以发现模型组动物在平台所在象限停留的时间明显短于对照组动物，但两组动物在穿越平台次数指标中没有明显的统计学差异，说明模型组动物的记忆功能也较差。见图2，在蛋白水平上的检测结果显示，模型组动物脑组织中的凋亡通路Caspase3酶的含量、脂质氧化产物丙二醛(MDA)含量以及谷胱甘肽过氧化物酶(GSH-px)含量均明显高于对照组动物，说明D-gal促进模型组动物脑神经细胞的凋亡、造成模型组动物脑组织脂质过氧化损伤，但两组动物脑组织中总的超氧化物歧化酶(SOD)含量并无明显差异。上述实验结果证明，采用D-gal制备的痴呆模型成功，D-gal可以损伤动物的学习和记忆功能。
2.雌性Metrnl敲除小鼠制备D-gal模型实验结果。见图3，从前五天的空间采集实验中可见，从第3天开始，两组呈现差别，其中第3天和第5天两组动物到达平台的潜伏期有统计学差异，KO组动物寻找平台的时间明显长于WT组动物，说明Metrnl敲除小鼠的学习能力比野生型小鼠较差；在第6天的探测试验中两组动物在平台所在象限停留的时间和穿越平台次数，在统计学上没有显著意义，但是穿越平台次数的均值在KO组显然较低，说明Metrnl敲除小鼠的记忆功能较野生型小鼠有所下降。
3.雄性Metrnl敲除小鼠制备D-gal模型实验结果。见图4。从前五天的空间采集实验中可见，从第3天开始，两组呈现差别，其中第3天和第5天两组动物到达平台的潜伏期有统计学差异，KO组动物寻找平台的时间明显长于WT组动物，与之前的雌性动物模型结论相吻合，说明Metrnl敲除小鼠的学习能力比野生型小鼠较差，没有性别差异；在第6天的探测试验中两组动物在平台所在象限停留的时间和穿越平台次数都没有明显的统计学差异，说明两组动物的记忆功能没有明显的差异。
4.雄性Metrnl敲除小鼠制备D-gal模型大脑海马区Western-blot实验结果。图5左图显示，在D-gal模型上，与野生型小鼠大脑海马区的蛋白表达水平相比，Metrnl敲除小鼠大脑海马区BDNF、GFAP和PSD95含量明显低于野生型对照小鼠，其中：BDNF是脑源性神经营养因子，与认知功能密切相关，PSD95是突触可塑性的标记物，而GFAP则是星形胶质细胞活化的标志，星形胶质细胞参与血脑屏障的构成。图5右图显示，在不造模条件下，上述这些蛋白表达在KO组与WT组之间没有明显差别。这些结果表明，在痴呆模型上，Metrnl蛋白可参与星形胶质细胞的活化及突触连接的形成，并调控脑源性神经营养因子的分泌，从而参与大脑认知功能的调控。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员，在不脱离本发明方法的前提下，还可以做出若干改进和补充，这些改进和补充也应视为本发明的保护范围。
Claims (5)
1.Metrnl蛋白或基因或它们的增效剂在制备防治认知功能障碍的药物中的应用。
2.Metrnl蛋白或基因或它们的增效剂在制备改善学习记忆能力的药物中的应用。
3.Metrnl蛋白或基因或它们的增效剂在制备改善认知功能障碍的食品和保健品中的应用。
4.Metrnl蛋白或基因或它们的增效剂在制备改善学习记忆能力的食品和保健品中的应用。
5.根据权利要求1-4任一所述的应用，其特征在于，所述的增效剂选自激动剂、上调剂或稳定剂。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810183174.4A CN110227146A (zh) | 2018-03-06 | 2018-03-06 | Metrnl蛋白或基因在防治认知障碍方面的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810183174.4A CN110227146A (zh) | 2018-03-06 | 2018-03-06 | Metrnl蛋白或基因在防治认知障碍方面的应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110227146A true CN110227146A (zh) | 2019-09-13 |
Family
ID=67862140
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810183174.4A Pending CN110227146A (zh) | 2018-03-06 | 2018-03-06 | Metrnl蛋白或基因在防治认知障碍方面的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110227146A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113462769A (zh) * | 2021-08-05 | 2021-10-01 | 中国医学科学院医药生物技术研究所 | 抑制剂/CaMKII体系及作为生物标志物中的应用 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102164611A (zh) * | 2008-07-24 | 2011-08-24 | Ns基因公司 | 生长因子metrnl的治疗用途 |
-
2018
- 2018-03-06 CN CN201810183174.4A patent/CN110227146A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102164611A (zh) * | 2008-07-24 | 2011-08-24 | Ns基因公司 | 生长因子metrnl的治疗用途 |
Non-Patent Citations (4)
| Title |
|---|
| JESPER ROLAND JØRGENSEN等: "Cometin is a novel neurotrophic factor that promotes neurite outgrowth and neuroblast migration in vitro and supports survival of spiral ganglion neurons in vivo", 《EXPERIMENTAL NEUROLOGY》 * |
| SI-LI ZHENG 等: "Metrnl: a secreted protein with new emerging functions", 《ACTA PHARMACOLOGICA SINICA》 * |
| SURACE C, PIAZZOLLA S,等: "Mild ring 17 syndrome shares common phenotypic features irrespective of the chromosomal breakpoints location", 《CLIN GENET》 * |
| 洪辰: "metrnl对小鼠认知功能的影响及机制研究", 《中国优秀硕士学位论文 医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113462769A (zh) * | 2021-08-05 | 2021-10-01 | 中国医学科学院医药生物技术研究所 | 抑制剂/CaMKII体系及作为生物标志物中的应用 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lu et al. | Depolarizing stimuli regulate nerve growth factor gene expression in cultured hippocampal neurons. | |
| Vaughan et al. | A morphometric study of light-induced damage in transgenic rat models of retinitis pigmentosa | |
| Foik et al. | Detailed visual cortical responses generated by retinal sheet transplants in rats with severe retinal degeneration | |
| KR101919081B1 (ko) | 인간 태반 추출물을 유효성분으로 포함하는 근 위축증 또는 근육감소증의 예방 또는 치료용 및 근육 기능 개선용 조성물 | |
| CN101466380A (zh) | 治疗帕金森病的方法和组合物 | |
| Hao et al. | CDKL5 deficiency augments inhibitory input into the dentate gyrus that can be reversed by deep brain stimulation | |
| Corey-Bloom et al. | Disease modifying potential of glatiramer acetate in Huntington's disease | |
| Borelli et al. | Distribution of Fos immunoreactivity in the rat brain after freezing or escape elicited by inhibition of glutamic acid decarboxylase or antagonism of GABA-A receptors in the inferior colliculus | |
| CN110227146A (zh) | Metrnl蛋白或基因在防治认知障碍方面的应用 | |
| O'Dell et al. | Disruption of rat deep cerebellar perineuronal net alters eyeblink conditioning and neuronal electrophysiology | |
| US20230346713A1 (en) | Methods and compositions for treating neurological conditions | |
| US10137156B2 (en) | Composition comprising neural cell and elastin-like polypeptide for treating Parkinson's disease | |
| CN115463218A (zh) | Shh通路调控生物节律及其相关应用 | |
| Fan et al. | Light prevents exogenous 11-cis retinal from maintaining cone photoreceptors in chromophore-deficient mice | |
| CN109675037B (zh) | Atp及其受体在制备治疗自闭症药物中的应用 | |
| Thompson et al. | Visual evoked potential evidence of albino-like chiasmal misrouting in a patient with Angelman syndrome with no ocular features of albinism | |
| Chang | Mouse models as tools to identify genetic pathways for retinal degeneration, as exemplified by Leber’s congenital amaurosis | |
| CN109223817A (zh) | 一种微小非编码rna的拮抗剂及其应用 | |
| CN102772418B (zh) | 富贵草胺碱g的用途 | |
| Ingensiep | Establishment of multielectrode array-based retinal ex vivo stress models: hypoxia and pressure | |
| CN106619590B (zh) | 一种治疗神经退行性疾病的药物组合物 | |
| Zhang et al. | Short-wavelength artificial light affects visual neural pathway development in mice | |
| Sun | Mitochondrial morphology and motility in retinal ganglion cells | |
| Evgenia | Enhancing visual cortical plasticity in mice by enriching their environment: a combined imaging and behavioural study | |
| Bhandare | Microglial and neurochemical mechanisms of seizure-induced central autonomic cardiovascular dysfunction |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220826 Address after: 200433 No. 800 Xiang Yin Road, Shanghai, Yangpu District Applicant after: PLA Naval Military Medical University Address before: Room 1A27, Building 3, No. 500, Songbin Road, Baoshan District, Shanghai, 200940 Applicant before: SHANGHAI FENGJING BIOMEDICAL TECHNOLOGY Co.,Ltd. |
|
| TA01 | Transfer of patent application right |