CN110754431A - Building method and application of osteoporosis and Alzheimer's disease combined animal model - Google Patents
Building method and application of osteoporosis and Alzheimer's disease combined animal model Download PDFInfo
- Publication number
- CN110754431A CN110754431A CN201911146361.6A CN201911146361A CN110754431A CN 110754431 A CN110754431 A CN 110754431A CN 201911146361 A CN201911146361 A CN 201911146361A CN 110754431 A CN110754431 A CN 110754431A
- Authority
- CN
- China
- Prior art keywords
- alzheimer
- osteoporosis
- disease
- animal model
- establishing
- 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
- 208000001132 Osteoporosis Diseases 0.000 title claims abstract description 82
- 208000024827 Alzheimer disease Diseases 0.000 title claims abstract description 79
- 238000010171 animal model Methods 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 26
- 239000003814 drug Substances 0.000 claims abstract description 34
- 229940079593 drug Drugs 0.000 claims abstract description 26
- FSBUXLDOLNLABB-ISAKITKMSA-N echinacoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FSBUXLDOLNLABB-ISAKITKMSA-N 0.000 claims abstract description 14
- NJYVDFDTLLZVMG-UHFFFAOYSA-N echinacoside Natural products CC1OC(OC2C(O)C(OCCc3ccc(O)c(O)c3)OC(COC4OC(CO)C(O)C(O)C4O)C2OC(=O)C=Cc5cc(O)cc(O)c5)C(O)C(O)C1O NJYVDFDTLLZVMG-UHFFFAOYSA-N 0.000 claims abstract description 14
- KDSWDGKIENPKLB-QJDQKFITSA-N verbascoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)CCC=2C=C(O)C(O)=CC=2)[C@@H](CO)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O KDSWDGKIENPKLB-QJDQKFITSA-N 0.000 claims abstract description 14
- QFRYQWYZSQDFOS-UHFFFAOYSA-N verbascoside Natural products CC1OC(COC2C(O)C(COC3OC(C(O)C(O)C3O)C(=O)O)OC(Oc4cc(O)cc5OC(=CC(=O)c45)c6ccc(O)c(O)c6)C2O)C(O)C(O)C1O QFRYQWYZSQDFOS-UHFFFAOYSA-N 0.000 claims abstract description 14
- 230000000857 drug effect Effects 0.000 claims abstract description 6
- 241000700159 Rattus Species 0.000 claims description 52
- 230000000694 effects Effects 0.000 claims description 42
- 210000000988 bone and bone Anatomy 0.000 claims description 39
- 210000001519 tissue Anatomy 0.000 claims description 27
- 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 claims description 22
- 238000001514 detection method Methods 0.000 claims description 20
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 claims description 17
- 238000004458 analytical method Methods 0.000 claims description 17
- 238000011156 evaluation Methods 0.000 claims description 17
- 238000002474 experimental method Methods 0.000 claims description 17
- 229940118019 malondialdehyde Drugs 0.000 claims description 17
- 102000019197 Superoxide Dismutase Human genes 0.000 claims description 16
- 108010012715 Superoxide dismutase Proteins 0.000 claims description 16
- 238000009806 oophorectomy Methods 0.000 claims description 16
- 230000002146 bilateral effect Effects 0.000 claims description 12
- 102000012440 Acetylcholinesterase Human genes 0.000 claims description 11
- 108010022752 Acetylcholinesterase Proteins 0.000 claims description 11
- 229940022698 acetylcholinesterase Drugs 0.000 claims description 11
- 238000003304 gavage Methods 0.000 claims description 10
- 238000012360 testing method Methods 0.000 claims description 10
- 238000012347 Morris Water Maze Methods 0.000 claims description 9
- RSEPBGGWRJCQGY-RBRWEJTLSA-N Estradiol valerate Chemical compound C1CC2=CC(O)=CC=C2[C@@H]2[C@@H]1[C@@H]1CC[C@H](OC(=O)CCCC)[C@@]1(C)CC2 RSEPBGGWRJCQGY-RBRWEJTLSA-N 0.000 claims description 8
- XWAIAVWHZJNZQQ-UHFFFAOYSA-N donepezil hydrochloride Chemical compound [H+].[Cl-].O=C1C=2C=C(OC)C(OC)=CC=2CC1CC(CC1)CCN1CC1=CC=CC=C1 XWAIAVWHZJNZQQ-UHFFFAOYSA-N 0.000 claims description 8
- 229960004766 estradiol valerate Drugs 0.000 claims description 8
- 230000000971 hippocampal effect Effects 0.000 claims description 8
- 239000007928 intraperitoneal injection Substances 0.000 claims description 8
- 238000013425 morphometry Methods 0.000 claims description 8
- 210000002966 serum Anatomy 0.000 claims description 8
- 238000010603 microCT Methods 0.000 claims description 7
- 230000003562 morphometric effect Effects 0.000 claims description 7
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims description 6
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims description 6
- 229960003135 donepezil hydrochloride Drugs 0.000 claims description 6
- 238000012795 verification Methods 0.000 claims description 6
- 238000011565 Alzheimer-disease animal model Methods 0.000 claims description 5
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 claims description 4
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 claims description 4
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 claims description 4
- 230000002378 acidificating effect Effects 0.000 claims description 4
- 238000007619 statistical method Methods 0.000 claims description 4
- 229940095064 tartrate Drugs 0.000 claims description 4
- 210000000683 abdominal cavity Anatomy 0.000 claims description 3
- 238000001543 one-way ANOVA Methods 0.000 claims description 3
- 238000012353 t test Methods 0.000 claims description 3
- 230000002496 gastric effect Effects 0.000 claims description 2
- 230000007246 mechanism Effects 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 230000002265 prevention Effects 0.000 abstract description 3
- 238000012216 screening Methods 0.000 abstract description 2
- 210000004291 uterus Anatomy 0.000 description 12
- 210000004556 brain Anatomy 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 201000010099 disease Diseases 0.000 description 7
- 210000001320 hippocampus Anatomy 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 210000004907 gland Anatomy 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 230000000877 morphologic effect Effects 0.000 description 6
- 230000037182 bone density Effects 0.000 description 5
- 210000000981 epithelium Anatomy 0.000 description 5
- 239000010410 layer Substances 0.000 description 5
- 210000002569 neuron Anatomy 0.000 description 5
- 210000001672 ovary Anatomy 0.000 description 5
- 210000002303 tibia Anatomy 0.000 description 5
- 210000000689 upper leg Anatomy 0.000 description 5
- 102100023460 Choline O-acetyltransferase Human genes 0.000 description 4
- 108010058699 Choline O-acetyltransferase Proteins 0.000 description 4
- 208000035896 Twin-reversed arterial perfusion sequence Diseases 0.000 description 4
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium chloride Substances Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 4
- 239000000890 drug combination Substances 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- -1 Estradiol Valerate Chemical compound 0.000 description 3
- 241000700157 Rattus norvegicus Species 0.000 description 3
- 229960004373 acetylcholine Drugs 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000007876 drug discovery Methods 0.000 description 3
- 210000004696 endometrium Anatomy 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000011633 osteoporosis animal model Methods 0.000 description 3
- 230000008506 pathogenesis Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 102000007591 Tartrate-Resistant Acid Phosphatase Human genes 0.000 description 2
- 108010032050 Tartrate-Resistant Acid Phosphatase Proteins 0.000 description 2
- 210000003489 abdominal muscle Anatomy 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 2
- 210000000577 adipose tissue Anatomy 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 230000007131 anti Alzheimer effect Effects 0.000 description 2
- 230000003262 anti-osteoporosis Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 229960003530 donepezil Drugs 0.000 description 2
- 238000007877 drug screening Methods 0.000 description 2
- 230000003937 effect on alzheimer disease Effects 0.000 description 2
- 230000002876 effect on osteoporosis Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 230000003387 muscular Effects 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 230000004792 oxidative damage Effects 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 208000001685 postmenopausal osteoporosis Diseases 0.000 description 2
- 238000011552 rat model Methods 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 230000009182 swimming Effects 0.000 description 2
- 238000010200 validation analysis Methods 0.000 description 2
- 208000010392 Bone Fractures Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 206010013710 Drug interaction Diseases 0.000 description 1
- 206010017076 Fracture Diseases 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
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 210000000702 aorta abdominal Anatomy 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000008416 bone turnover Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000004349 growth plate Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000007786 learning performance Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007087 memory ability Effects 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 210000002682 neurofibrillary tangle Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000004154 testing of material Methods 0.000 description 1
- 210000001694 thigh bone Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/25—Animals on a special diet
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/30—Animals modified by surgical methods
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0306—Animal model for genetic diseases
- A01K2267/0312—Animal model for Alzheimer's disease
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The application provides a method for establishing an animal model with osteoporosis and Alzheimer's disease and application thereof3And obtaining the animal model of osteoporosis combined with Alzheimer's disease. The application is the application of the establishment method of the animal model with osteoporosis and Alzheimer's disease in verifying the drug effect of the drug, wherein the drug comprises one or more of the drugs for treating osteoporosis, the drugs for treating Alzheimer's disease, echinacoside and verbascoside. The model has the advantages of short experimental period, high success rate, low cost and easy copying, application and popularization; the animal model of osteoporosis and Alzheimer's disease can be used as a research object to explore the complication mechanism, early prevention and simultaneous treatment of osteoporosis and Alzheimer's disease, and can also be used for discovery and screening of a multi-effect drug.
Description
Technical Field
The invention relates to an animal model establishing method and application thereof, in particular to an animal model establishing method for osteoporosis combined Alzheimer's disease.
Background
Osteoporosis (OP) and Alzheimer's Disease (AD) are the most common clinical senile diseases and are frequently complicated, and their complicated pathological mechanisms always limit effective prevention and treatment of the two diseases. AD is a degenerative disease of the nervous system characterized primarily by the formation of age spots, neurofibrillary tangles and neuronal loss. OP is a systemic bone disease characterized by decreased bone density and deterioration of the microstructure of bone tissue, resulting in increased bone fragility and increased risk of fracture. With the increasing aging of population, OP and AD become important public health problems facing China. The total disease rate of OP in China is 13% on average, the AD disease rate can reach 5-7%, and the tendency of year-by-year rising is presented, and the elderly and women are susceptible to both diseases. Increasing research has shown that there are several common pathogenic factors and pathogenesis of OP and AD. The traditional Chinese medicine theory considers that the kidney governs bones, produces marrow and leads to the brain, the pathogenesis of AD and OP in traditional Chinese medicine is closely related, the pathogenesis of 'deficiency of kidney essence, no marrow production of essence and marrow loss is embodied in two diseases', and the possibility of 'treating the abnormal diseases simultaneously' is suggested.
In clinic, OP and AD are mainly treated by drug combination, but generally have the problems of unsatisfactory curative effect, large dose, strong toxic and side effects, unknown drug interaction and the like.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a method for establishing an animal model with osteoporosis and Alzheimer's disease; the second purpose of the invention is to provide the application of the establishing method of the osteoporosis and Alzheimer's disease animal model in verifying the drug effect of the drug.
The technical scheme is as follows: the invention provides a method for establishing an animal model with osteoporosis and Alzheimer's disease, wherein rats subjected to bilateral ovary extirpation are used as experimental objects, D-galactose is injected into abdominal cavities, and AlCl is perfused into stomachs at the same time3And obtaining the animal model of osteoporosis combined with Alzheimer's disease.
Preferably, the intraperitoneal injection of D-galactose is initiated at week 4 after bilateral ovariectomy.
Further, the dose of the D-galactose for intraperitoneal injection is 150 mg/kg/day; intragastric AlCl3The dose of (A) was 30 mg/kg/day for 8 weeks.
Preferably, the steps further comprise one or both of alzheimer model evaluation comprising Morris water maze test and determination of one or more of acetylcholinesterase activity, superoxide dismutase activity and malondialdehyde content in hippocampal tissues and osteoporosis model evaluation or intervention validation of an osteoporosis-combined alzheimer animal model;
the osteoporosis model evaluation comprises one or more of measuring the contents of bone-specific alkaline phosphatase activity, tartrate-resistant acidic phosphatase activity, superoxide dismutase activity and malondialdehyde in serum and one or more of uterine tissue morphology observation, morphometric detection, bone biomechanical detection and Micro-CT detection and analysis.
Wherein, acetylcholinesterase is called AChE for short; the acetylcholine transferase is abbreviated as ChAT; superoxide dismutase (SOD) for short; malondialdehyde is abbreviated as MDA; bone-specific alkaline phosphatase, BALP for short; tartrate-resistant acid phosphatase is abbreviated as TRAP.
Further, the intervention is verified by respectively applying estradiol valerate and donepezil hydrochloride to the osteoporosis combined Alzheimer's disease animal model, and performing Alzheimer's disease model evaluation and osteoporosis model evaluation; the Alzheimer disease model evaluation comprises a Morris water maze experiment and determination of one or more of acetylcholinesterase activity, superoxide dismutase activity and malondialdehyde content in hippocampal tissues;
the osteoporosis model evaluation comprises one or more of measuring the contents of bone-specific alkaline phosphatase activity, tartrate-resistant acidic phosphatase activity, superoxide dismutase activity and malondialdehyde in serum and one or more of uterine tissue morphology observation, morphometric detection, bone biomechanical detection and Micro-CT detection and analysis.
Further, the estradiol valerate was administered at 0.6 mg/kg/day and donepezil hydrochloride at 0.8 mg/kg/day at the beginning of gavage at 4 weeks after the bilateral ovariectomy.
Preferably, the gavage administration is for 8 weeks.
The invention also provides application of the establishing method of the osteoporosis and Alzheimer disease animal model in verifying drug effect, wherein the drugs comprise one or more of osteoporosis drugs, Alzheimer disease drugs, echinacoside and verbascoside.
Preferably, the osteoporosis drug, the Alzheimer disease drug, the echinacoside and/or the verbascoside are/is 0-10 parts of echinacoside and 10-0 parts of verbascoside.
Wherein, the echinacoside is ECH for short; the verbascoside is abbreviated ACT.
Preferably, the verification of the drug effect is a group verification, wherein the group verification comprises that estradiol valerate is dosed at 0.6 mg/kg/day, donepezil hydrochloride is dosed at 0.8 mg/kg/day, echinacoside is dosed at 90 mg/kg/day, verbascoside is dosed at 90 mg/kg/day, or echinacoside and verbascoside with the mass ratio of (1-10) and (10-1) are respectively administered by gastric gavage to rats in the experimental group from the 4 th week after bilateral ovariectomy, and the results are detected and statistically analyzed.
Preferably, the administration by gavage is continuous for 8 weeks.
Furthermore, the statistical analysis data result is that two groups are compared by adopting a t test, a plurality of groups are compared by adopting One-way ANOVA test for analysis, two groups are compared by adopting Tukey's multiple comparison, data are analyzed by adopting One or more of Graph Pad Prism5 software enzymatic activity and malondialdehyde content, and uterine tissue morphology observation and morphology measurement, and the difference with the set P <0.05 has significance.
Has the advantages that: 1. the method comprises the steps of firstly, constructing an Alzheimer disease model based on a classical ovariectomized osteoporosis rat model to obtain an osteoporosis combined Alzheimer disease animal model; then performing Morris water maze experiment, serum biochemical index analysis, brain hippocampal tissue biochemical index analysis and morphological analysis, uterus tissue morphological analysis, bone biomechanical detection, and bone Micro-CT analysisAnalyzing and other detection indexes, and the application is based on combined intraperitoneal injection of D-galactose and gavage AlCl for rat ovariectomy3The modeling method of (2) constructs a rat model of osteoporosis combined with Alzheimer's disease, and the model has the advantages of short experimental period, high success rate, low cost, easy replication and convenient application and popularization;
2. the animal model of osteoporosis combined with Alzheimer disease constructed by the application is evaluated, and double-effect medicines which are simultaneously anti-osteoporosis and anti-Alzheimer disease are discovered and screened by taking the model as an index, so that a research foundation and a data support are provided for discovering multi-effect active monomers from traditional Chinese medicines and discussing a mechanism of 'treating different diseases simultaneously', wherein the multi-effect active monomers including echinacoside and verbascoside have obvious pharmacological activity on bones and a nervous system, and show the effects of resisting osteoporosis of rats caused by ovariectomy and dementia of rats caused by bilateral ventricular injection A β;
3. the application meets the relevant requirements of the animal model of osteoporosis and Alzheimer's disease at the same time, not only can the animal model of osteoporosis and Alzheimer's disease be used as a research object to explore the complication mechanism, early prevention and simultaneous treatment of osteoporosis and Alzheimer's disease, but also can be used for discovering and screening multiple-effect medicines of one medicine.
Drawings
FIG. 1 is a schematic diagram of the method for establishing an animal model with osteoporosis and Alzheimer's disease and the pharmaceutical intervention thereof, provided by the invention;
FIG. 2 is a structural diagram of potential dual effect anti-osteoporosis and anti-Alzheimer's disease compounds provided by the present invention;
FIG. 3 shows the uterine tissue morphology of rats in each group according to the embodiment of the present invention, wherein n is 3, LP represents the inherent layer of uterus, EP represents the uterine mucosal epithelium, ▲ represents the uterine gland, the uterus of Sham group is the thick-walled muscular organ, the uterine mucosal epithelium is the single-layer columnar epithelium, the inherent layer is distributed with uterine glands and blood vessels, the uterus of OVX group, OVX + D group is obviously shriveled, the lumen is narrowed, the endometrium is obviously thinned, the number of uterine glands is reduced, particularly OVX + D group is more obvious, the uterine tissue morphology of ECH group, ACT group and P1 group is obviously improved compared with that of OVX + D group, the uterus is obviously filled, the endometrium is obviously thickened, and the number of uterine glands is increased.
Detailed Description
In the following, embodiments of the present invention will be described in detail with reference to the accompanying drawings, and unless otherwise specified, the reagents used in the present invention are commercially available.
Example one
Construction of osteoporosis and Alzheimer's disease animal model
1. Establishment of postmenopausal osteoporosis model
8-week-old female Wistar rats, SPF grade, were subjected to bilateral ovariectomy after 1-week acclimatization feeding. Rats after fixed anesthesia in prone position were prepared on both sides of the back. Firstly, a small opening is cut through a pair of surgical scissors from one side of the back and abdomen, the skin, the abdominal muscle and the peritoneum are cut in sequence, then the adipose tissues are stripped to find out a red-brown vegetable-shaped ovary, the ovary is removed after the uterine horn is ligated by a surgical suture line, then the abdominal muscle and the skin are sutured in sequence, and the wound is externally coated with penicillin powder. The other ovary was removed in the same way. The sham group removed only the same volume of adipose tissue near the ovaries, and the other procedures were identical. After the operation is finished, the animal is raised for 3 weeks.
2. Establishment of Alzheimer's disease model
In the establishment of the postmenopausal osteoporosis model, the raised rats are grouped as follows: sham group (Sham group), independent ovariectomy model group (OVX group), independent D-galactose intraperitoneal injection combined intragastric perfusion AlCl3Model group, namely group D, ovariectomy and D-galactose intraperitoneal injection combined intragastric AlCl3Model groups, i.e., OVX + D groups, were 9 per group.
The model for Alzheimer's disease was established starting at week 4 after bilateral ovariectomy. D-galactose is injected into abdominal cavity at a dose of 150mg/kg/D, and AlCl is infused into stomach3The dose was 30mg/kg/d for 8 weeks.
3. Alzheimer's disease model evaluation
3.1Morris Water maze experiment
After molding for 8 weeks, the learning and memory abilities of the rats were measured using the Morris water maze. A cylindrical hiding platform is placed in the center of 1 quadrant of 4 quadrants of the Morris water maze optionally, and the position of the platform is kept unchanged all the time during the whole experiment. The water level in the water maze is 1cm higher than the hidden platform, so that the hidden platform cannot be seen directly on the water level, and the water temperature is controlled to be (26 +/-1) DEG C. The video recorder is installed above the water maze and is connected with the computer, and the condition of the platform is searched and analyzed by the automatically recorded rat.
Positioning navigation test: the duration is 5 days, the rats are trained for 4 times in a fixed time period every day, and the time required for the rats to find the hidden platform is recorded, namely the escape latency. If the rat does not find a hidden platform within 60s, it is directed by the experimenter to the hidden platform, its latency is recorded as 60s and the platform dwell time is 10 s. The average of 4 training latencies per day was taken as the learning performance of the rat on the current day.
And (3) space exploration test: and (4) removing the platform after the positioning navigation test is finished, putting the rat into water from the alignment quadrant of the target quadrant where the platform is located, and recording the times of crossing the position of the original platform within 60s and the activity time of the rat in the target quadrant.
Wherein, the escape latency in each group of rats in the positioning navigation test is compared and shown in table 1; the number of platform crossings and target quadrant residence times in the spatial exploration experiments for each group of rats are shown in table 2.
TABLE 1 comparison of escape latencies in localized voyage experiments for groups of rats described in the examples of the present invention
Note: data are expressed as mean ± standard deviation (n ═ 8) in seconds(s). Compared with the group of the Sham,#P<0.05，##P<0.01; compared with the group ratio of OVX + D,*P<0.05，**P<0.01。
TABLE 2 number of passes across the platform and residence time in the target quadrant for each group of rats in the space exploration test described in the examples of the present invention
Note: data are expressed as mean ± standard deviation (n ═ 8). Compared with the group of the Sham,#P<0.05，##P<0.01; compared with the group ratio of OVX + D,*P<0.05。
3.2 Biochemical index detection
After anaesthetizing, rats quickly pick up the brain on ice, peel off the cortex and hippocampus, place in an EP tube without RNase, and store at-80 ℃. According to the ELISA kit specification, the acetylcholinesterase (AChE) activity, the acetylcholinesterase (ChAT) activity, the superoxide dismutase (SOD) activity and the Malondialdehyde (MDA) content in the hippocampal tissues are measured.
3.3 Hippocampus morphological Observation
After anaesthetizing, rats are perfused to take out the brain, and placed in tissue fixing liquid for fixing and storing. Taking a picture of the stained section of the coronal section HE of the brain tissue by using an inverted fluorescence microscope, and observing the morphological changes of a CA1 area, a CA2 area, a CA3 area and a DG area of the hippocampal tissue with emphasis.
4. Osteoporosis model evaluation
4.1 Biochemical index detection
After rat anesthesia, blood is taken from abdominal aorta, the rat is kept still for 2-3 h at room temperature, then the rat is centrifuged for 10min at 3500r, and serum is collected and subpackaged and stored at minus 80 ℃. The contents of bone-specific alkaline phosphatase (BALP activity), tartrate-resistant acid phosphatase (TRAP activity), superoxide dismutase (SOD activity) and Malondialdehyde (MDA) in serum were determined according to the ELISA kit instructions.
4.2 morphological Observation and morphometric detection of uterine tissue
After anaesthetizing, the rat takes out the uterus, cleans the uterus with normal saline, sucks excessive moisture, weighs, records the wet weight of the uterus, and then places the uterus in tissue fixing liquid for fixed storage for HE staining. Taking a picture and observing the stained section of the uterine cross section HE by adopting an inverted fluorescence microscope, randomly selecting 5 visual fields respectively, carrying out morphological measurement on the diameter of the uterine cross section and the thickness of uterine mucosa epithelium by using Image-Pro Plus 6.0 software, and counting the number of the inherent layer uterine glands.
4.3 Micro-CT detection
After anesthetizing, the rat takes the left femur, the right femur and the tibia along the joints, removes the attached muscle and connective tissue, fixes and stores the right femur with tissue fixing liquid, each group comprises 3, scans along the long axis direction of the femur by using Micro-CT, and the scanning resolution is 18 μm. And after the scanning is finished, performing three-dimensional reconstruction on the sample, and introducing the reconstructed sample into CTAn software. The growth plate is used as a reference surface, a position 1mm below the reference surface is selected as an upper limit, 100 layers are selected downwards to be used as VOI, a three-dimensional reconstruction diagram is directly used for measuring a trabecular bone micro-tissue structure, bone density values and bone shape measurement parameters are obtained, wherein the bone mass percentage is bone volume/tissue volume, BV/TV, the number of trabecular bone beams is trabecular number, Tb.N, the thickness of the trabecular bone beams is trabecular thickness, Tb.Th, trabecular bone gap, Tb.Sp, structural model index, SMI, bone surface density is bone surface/tissue volume, BS/TV and trabecular bone mode factor are trabecular pattern factor, Pf..
4.4 biomechanical testing
Taking out the right tibia samples frozen at-80 ℃, and soaking 4 tibia samples in physiological saline for 3-4 hours. The right tibia full section is taken and tested on a universal material testing machine. The sample was placed on two supporting bars with a distance of 25mm between the bars and the physiological curvature of the sample was upward. The loading rod of the load is positioned in the center of the specimen, the specimen is loaded at a constant speed at a loading rate of 1.5mm/min until the specimen is broken, and a mechanical experiment tester records a load-displacement curve with software and analyzes data. The biomechanical parameters measured include maximum load, abbreviated as N, energy absorption, abbreviated as J, maximum stress, abbreviated as MPa.
5. Statistical analysis
The experimental data are expressed by mean +/-standard deviation, t test is adopted for comparison between two groups, One-way ANOVA test is adopted for comparison between multiple groups for analysis, Tukey's multiple comparison is adopted for pairwise comparison, Graph Pad Prism5 software is adopted for analysis, and the difference with the set P <0.05 is significant.
6. Results of the experiment
Compared with the Sham group, the wet weight of the uterus of the rats in the OVX + D group is obviously reduced, namely P is less than 0.01, the uterus of the rats in the OVX group and the OVX + D group is obviously shrunk, the lumen is narrowed, the endometrium and the muscular layer are obviously thinned, the diameter of the transverse section of the uterus, the thickness of the epithelium of the uterine mucous membrane and the quantity of uterine glands are obviously reduced, namely P is less than 0.01, and P is less than 0.001, thus prompting the success of the bilateral ovariectomy operation. Compared with the Sham group, the activity of BALP and TRAP of the OVX group and the OVX + D group is obviously increased, namely P is less than 0.001, and the activity is typical of the high bone turnover osteoporosis caused by ovariectomy. Bone histopathology and bone morphometry parameters show that the rat femoral cancellous bone density of the OVX group and the rat of the OVX + D group is reduced, the tibial trabecular bone is broken, the trabecular space is widened, the structure is disordered, and bone trabecular related parameters such as the bone mass percentage, the bone trabecular thickness and the like are obviously reduced compared with the Sham group. In addition, bone biomechanical results show that compared with the Sham group, the maximum load and the maximum stress borne by the OVX + D group are both obviously reduced, namely P is less than 0.05, and the energy absorption is also greatly reduced. These results all suggest that the model animal has the pathological change of osteoporosis, which indicates that the osteoporosis model is successfully constructed.
The Morris water maze behavioural experiment found that the mean latency of OVX + D group rats was much higher than that of Sham group from day 2. Compared with the Sham group, the number of times of crossing the platform and the swimming time in the target quadrant of the rats in the D group and the OVX + D group are obviously reduced within 60s, namely P is less than 0.05, and P is less than 0.01, which indicates that the symptoms of the Alzheimer disease of the model animals are changed. The analysis result of the biochemical indexes of the brain hippocampus tissue shows that the AChE activity of the OVX + D group is obviously improved, namely P is less than 0.05, which shows that the hydrolysis of neurotransmitter acetylcholine in the brain of rats in the group is accelerated; ChAT activity was significantly decreased in all of OVX, D and OVX + D groups, i.e., P <0.05 and P <0.001, indicating that synthesis of acetylcholine, a neurotransmitter in the brain, was slowed. In addition, compared with the normal neuron cells of the Sham group rats, the number of the neuron cells in each region of the hippocampus of the rats in the OVX group, the rats in the D group and the OVX + D group is obviously reduced, the cell arrangement is disordered and the gap is increased, and particularly, the number of the neuron cells in the hippocampus of the rats in the D group and the rats in the OVX + D group is less, and the neuron cells are disorderly scattered and distributed, the cell nucleus is fixed and contracted, the cell morphology is not obvious, and the neuron cells are. The results show that the model animal has obvious Alzheimer disease pathological changes, and the construction of the Alzheimer disease model is successful.
In conclusion, the animal model for osteoporosis combined with Alzheimer disease is successfully constructed.
Wherein, the change of uterine wet weight of each group of rats is shown in a table 4:
TABLE 4 uterine wet weight changes in groups of rats described in the examples of the present invention
Note: data are expressed as mean ± standard deviation (n ═ 6). Compared with the group of the Sham,##P<0.01。
example two
Intervention validation in animal models of osteoporosis and alzheimer's disease
1. Experimental methods
An animal model of Wistar rat osteoporosis combined with Alzheimer's disease is established according to the method of the embodiment, Estradiol Valerate, namely Estradiol Valerate, which is a classic drug for treating osteoporosis in clinic, EV for short, and Donepezil HCl, namely Donepezil HCL, which is a classic drug for treating Alzheimer's disease in clinic, are respectively applied to the animal model of the invention, DPZ for short. Wherein EV is administrated by gavage at the dose of 0.6mg/kg/d and DPZ at the dose of 0.8mg/kg/d in the stage of Alzheimer disease molding. After 8 weeks of continuous administration, the quantitative indicators of osteoporosis and Alzheimer's disease were determined according to the first example of the present invention.
2. Results of the experiment
In the Morris water maze experiment, compared with the OVX + D group, the incubation periods of the EV group and the DPZ group are obviously shortened, namely P is less than 0.05, P is less than 0.01, the platform crossing times and the swimming time of a target quadrant are increased to different degrees, and the part has significant difference, namely P is less than 0.05. Compared with OVX + D group, the activity of the classic osteoporosis indexes of BALP and TRAP and the activity of the classic Alzheimer disease indexes of AChE and ChAT are obviously improved, namely P is less than 0.05. The drug dry prognosis, uterine morphology, bone tissue morphology and bone biomechanical parameters of rats in the EV group, neuronal morphology of each region of the hippocampus of rats in the DPZ group and the like all show improvement in different degrees.
The results of the embodiment show that the two classic drugs can respectively and obviously improve the symptoms of osteoporosis and Alzheimer's disease of the osteoporosis and Alzheimer's disease combined animal model of the invention, and prove that the model of the osteoporosis and Alzheimer's disease combined animal model of the invention is successfully made, and simultaneously, the possibility that the model is used for double-effect drug discovery and drug screening for resisting osteoporosis and Alzheimer's disease is also prompted.
EXAMPLE III
Double-effect drug discovery of echinacoside, verbascoside and combination thereof based on animal model of osteoporosis combined with Alzheimer's disease
1. Experimental methods
Animal models of Wistar rat osteoporosis combined with Alzheimer's disease were established as described in example one, and EV and DPZ were administered to the animal models of the present invention as two positive control groups, respectively, as described in example two. Candidate drugs echinacoside, namely ECH group, verbascoside, namely ACT group, and two monomer drug combination, namely P1 group are respectively applied to the osteoporosis and Alzheimer disease animal model. Wherein, ECH group is administrated by intragastric gavage at the dose of 90mg/kg/d, ACT group is administrated at the dose of 90mg/kg/d, and P1 group is administrated at the dose of ECH and ACT 1:1 ratio at the model building stage of Alzheimer disease. The two positive control groups were dosed and timed according to example two. After 8 weeks of continuous administration, the quantitative indicators of osteoporosis and Alzheimer's disease were examined according to the first example of the present invention. Compared with the classical medicines for treating osteoporosis and Alzheimer's disease, the effect of ECH, ACT and the combination thereof on the quantitative indexes of the osteoporosis and the Alzheimer's disease is compared, and the effect of the potential double-effect medicine and the medicine combination thereof is evaluated.
2. Results of the experiment
Ovariectomy and intraperitoneal injection of D-galactose combined intragastric AlCl3Can cause large-scale oxidative damage to rats and reduce SOD activity thereofThe MDA content is increased; after ECH and ACT are dried, the SOD activity in the blood serum and the brain hippocampus tissue is obviously increased, namely P<0.05，P<0.001 and simultaneously the MDA content is also obviously reduced, namely P<0.05，P<0.001, suggesting that both monomers may show related efficacy by alleviating oxidative damage in model animals. According to the results of biochemical index analysis, brain hippocampal tissue morphology analysis, uterine tissue morphology analysis, bone morphology analysis and bone biomechanical detection experiments, compared with OVX + D group, ECH and ACT can simultaneously improve the symptoms and quantitative indexes of osteoporosis and Alzheimer's disease of model animals to a certain extent after single or combined treatment, and show the curative effects similar to those of classical drugs EV and DPZ. Especially, the ECH and ACT have better relieving effect on osteoporosis and Alzheimer disease when being intervened by drug combination, and the effect is better than that of a single drug.
The embodiment proves that the animal model constructed in the invention can be used for simultaneously improving the feasibility of double-effect and multi-effect drug discovery and drug screening of osteoporosis and Alzheimer's disease. The candidate drugs ECH, ACT and the combination thereof can show the improvement effect on osteoporosis and Alzheimer disease related quantitative indexes of an osteoporosis and Alzheimer disease animal model to different degrees, and the ECH, ACT and the combination thereof are proved to have the double-effect of resisting osteoporosis and Alzheimer disease. In addition, the invention also finds that the drug effect of the drug combination of echinacoside and verbascoside is better than that of a single drug, and suggests that the combination of the two compounds with different proportions can have the functions of decrement and synergy.
Wherein the morphometric determination of the uterine tissue of the rats in each group is shown in Table 5:
TABLE 5 morphometric determination of uterine tissue in groups of rats described in the examples of the invention
Note: data are expressed as mean ± standard deviation (n ═ 5). Compared with the group of the Sham,##P<0.01，###P<0.001; compared with the group ratio of OVX + D,*P<0.05。
the measurement of the bone density and bone trabecular morphological parameters of the thighbone of the rats in each group is shown in the table 6:
TABLE 6 measurement of bone Density and trabecular bone morphology parameters of femurs of rats of the groups described in the examples of the present invention
Note: data are expressed as mean ± standard deviation (n ═ 3). Compared with the group of the Sham,#P<0.05，##P<0.01; compared with the group ratio of OVX + D,*P<0.05。
the biomechanical parameters of the tibia of the rats in each group are determined as shown in the table 7:
TABLE 7 measurement of tibial biomechanical parameters for various groups of rats described in the examples of the invention
Note: data are expressed as mean ± standard deviation (n ═ 4). Compared with the group of the Sham,#P<0.05，###P<0.001; compared with the group ratio of OVX + D,*P<0.05。
Claims (10)
1. a method for establishing an animal model with osteoporosis and Alzheimer's disease, which is characterized by comprising the following steps: the method takes a rat subjected to bilateral ovariectomy as an experimental object, D-galactose is injected into the abdominal cavity, and AlCl is perfused into the stomach3And obtaining the animal model of osteoporosis combined with Alzheimer's disease.
2. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 1, wherein: the intraperitoneal injection of the D-galactose is started at the 4 th week after bilateral ovariectomy.
3. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 1, wherein: the dose of the D-galactose for intraperitoneal injection is 150 mg/kg/day; gavage stomachAlCl3The dose of (A) was 30 mg/kg/day for 8 weeks.
4. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 1, wherein: the step also comprises one or two of Alzheimer disease model evaluation and osteoporosis model evaluation or intervention verification of an osteoporosis combined Alzheimer disease animal model, wherein the Alzheimer disease model evaluation comprises a Morris water maze experiment and determination of one or more of acetylcholinesterase activity, superoxide dismutase activity and malondialdehyde content in hippocampal tissues;
the osteoporosis model evaluation comprises one or more of measuring the contents of bone-specific alkaline phosphatase activity, tartrate-resistant acidic phosphatase activity, superoxide dismutase activity and malondialdehyde in serum and one or more of uterine tissue morphology observation, morphometric detection, bone biomechanical detection and Micro-CT detection and analysis.
5. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 3, wherein: the intervention verification is that estradiol valerate and donepezil hydrochloride are respectively applied to the osteoporosis combined Alzheimer's disease animal model, and Alzheimer's disease model evaluation and osteoporosis model evaluation are carried out; the Alzheimer disease model evaluation comprises a Morris water maze experiment, and one or more of acetylcholinesterase activity, superoxide dismutase activity and malondialdehyde content in hippocampal tissues are measured;
the osteoporosis model evaluation comprises one or more of measuring the contents of bone-specific alkaline phosphatase activity, tartrate-resistant acidic phosphatase activity, superoxide dismutase activity and malondialdehyde in serum and one or more of uterine tissue morphology observation, morphometric detection, bone biomechanical detection and Micro-CT detection and analysis.
6. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 4, wherein: the estradiol valerate was dosed at 0.6 mg/kg/day and donepezil hydrochloride at 0.8 mg/kg/day, beginning gavage at week 4 after bilateral ovariectomy.
7. The method for establishing an animal model of osteoporosis combined with alzheimer's disease according to claim 5, wherein: the gavage administration is continued for 8 weeks.
8. The use of the method of establishing an animal model of osteoporosis and alzheimer's disease as set forth in claim 1 for verifying the efficacy of a drug, wherein: the medicine comprises one or more of osteoporosis treatment medicine, Alzheimer disease treatment medicine, echinacoside and verbascoside.
9. The use of the method for establishing an animal model of osteoporosis and alzheimer's disease according to claim 7 for verifying the efficacy of a drug, wherein: the drug effect of the verification drug is that the rats in the experimental group are respectively subjected to gastric lavage administration, detection and statistical analysis of data results at the 4 th week after bilateral ovariectomy, wherein the dose of estradiol valerate is 0.6 mg/kg/day, the dose of donepezil hydrochloride is 0.8 mg/kg/day, the dose of echinacoside is 90 mg/kg/day, the dose of verbascoside is 90 mg/kg/day, or the mass ratio of echinacoside and verbascoside is (1-10) (10-1).
10. The method for establishing an animal model of osteoporosis complicated with alzheimer's disease according to claim 9, wherein: the statistical analysis data result is that two groups are compared by adopting t test, the multiple groups are compared by adopting One-way ANOVA test for analysis, two-two comparison adopts Tukey's multiple comparison, data are analyzed by adopting Graph Pad Prism5 software, and the difference with the set P <0.05 has significance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911146361.6A CN110754431B (en) | 2019-11-21 | 2019-11-21 | Building method and application of osteoporosis and Alzheimer's disease combined animal model |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911146361.6A CN110754431B (en) | 2019-11-21 | 2019-11-21 | Building method and application of osteoporosis and Alzheimer's disease combined animal model |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110754431A true CN110754431A (en) | 2020-02-07 |
| CN110754431B CN110754431B (en) | 2021-09-28 |
Family
ID=69338839
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911146361.6A Active CN110754431B (en) | 2019-11-21 | 2019-11-21 | Building method and application of osteoporosis and Alzheimer's disease combined animal model |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110754431B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113440534A (en) * | 2020-03-26 | 2021-09-28 | 上海医药工业研究院 | Application of verbascoside in preparation of medicines |
| CN114159454A (en) * | 2021-12-16 | 2022-03-11 | 中国药科大学 | Traditional Chinese medicine composition for preventing and treating Alzheimer disease and/or osteoporosis, preparation method and application |
| CN114158524A (en) * | 2021-12-28 | 2022-03-11 | 上海市中医老年医学研究所 | Method for constructing atherosclerosis and osteoporosis comorbid animal model |
| CN114794014A (en) * | 2022-06-09 | 2022-07-29 | 辽宁中医药大学 | Animal model for establishing Alzheimer disease yin deficiency syndrome and/or Alzheimer disease non-yin deficiency syndrome by coprophilous fungus transplantation method, evaluation and application |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103636555A (en) * | 2013-11-11 | 2014-03-19 | 徐又佳 | Method for establishing mouse castrating high-ferrum osteoporotic model |
| WO2016137841A1 (en) * | 2015-02-26 | 2016-09-01 | William Paterson University Of New Jersey | Transgenic mice |
| CN107760713B (en) * | 2016-08-23 | 2020-05-05 | 中国科学院上海药物研究所 | Method for establishing animal model of neuropsychiatric disease of non-human mammal and application thereof |
| CN107197828A (en) * | 2017-07-07 | 2017-09-26 | 四川普莱美生物科技集团有限公司 | A kind of construction method of new osteoporosis model and the raising formula of use |
-
2019
- 2019-11-21 CN CN201911146361.6A patent/CN110754431B/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113440534A (en) * | 2020-03-26 | 2021-09-28 | 上海医药工业研究院 | Application of verbascoside in preparation of medicines |
| CN114159454A (en) * | 2021-12-16 | 2022-03-11 | 中国药科大学 | Traditional Chinese medicine composition for preventing and treating Alzheimer disease and/or osteoporosis, preparation method and application |
| CN114158524A (en) * | 2021-12-28 | 2022-03-11 | 上海市中医老年医学研究所 | Method for constructing atherosclerosis and osteoporosis comorbid animal model |
| CN114158524B (en) * | 2021-12-28 | 2023-03-10 | 上海市中医老年医学研究所 | Construction method of atherosclerosis and osteoporosis co-morbidity animal model |
| CN114794014A (en) * | 2022-06-09 | 2022-07-29 | 辽宁中医药大学 | Animal model for establishing Alzheimer disease yin deficiency syndrome and/or Alzheimer disease non-yin deficiency syndrome by coprophilous fungus transplantation method, evaluation and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110754431B (en) | 2021-09-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110754431B (en) | Building method and application of osteoporosis and Alzheimer's disease combined animal model | |
| Goettsch et al. | A single injection of gain-of-function mutant PCSK9 adeno-associated virus vector induces cardiovascular calcification in mice with no genetic modification | |
| Tian et al. | Efficacy of novel highly specific bromodomain-containing protein 4 inhibitors in innate inflammation–driven airway remodeling | |
| Yu et al. | Kinetic alterations of collagen and elastic fibres and their association with cardiac function in acute myocardial infarction | |
| Chen et al. | Establishment of the concurrent experimental model of osteoporosis combined with Alzheimer's disease in rat and the dual-effects of echinacoside and acteoside from Cistanche tubulosa | |
| Das et al. | Syndecan-4 enhances therapeutic angiogenesis after hind limb ischemia in mice with type 2 diabetes | |
| CN109600992A (en) | The method for treating or preventing Alzheimer disease and related conditions | |
| CN110520112A (en) | Dominant activity Yap as Hippo effector induces chromatin accessibility and cardiac muscle cell to update | |
| Liquitaya-Montiel et al. | Insulin growth factor-I promotes functional recovery after a focal lesion in the dentate gyrus | |
| CN110755450B (en) | Application of extracellular vesicles derived from mesenchymal stem cells in treatment of subarachnoid hemorrhage | |
| CN108785308B (en) | application of antagonist of nuclear receptor Rev-erb α in preparation of anti-abdominal aortic aneurysm drugs | |
| Qin et al. | Cardioprotective effect of ultrasound‐targeted destruction of Sirt3‐loaded cationic microbubbles in a large animal model of pathological cardiac hypertrophy | |
| Epstein et al. | Disintegration and esophageal irritation profiles of alendronate formulations: implications for clinical safety and efficacy | |
| CN110251677A (en) | For treating the pharmaceutical composition and its application of pulmonary fibrosis | |
| CN110794143B (en) | Method for researching interaction between adjacent cells | |
| CN110511930B (en) | Sal-miR-58 and application thereof in inhibition of vascular inflammatory reaction and aneurysm formation | |
| CN111647640A (en) | Method for rapidly and accurately realizing classification of cardiac function and course of chronic heart failure | |
| CN111686119A (en) | Traditional Chinese medicine monomer for treating early endometriosis and application thereof | |
| Zolfaghari et al. | microRNA-96 targets the INS/AKT/GLUT4 signaling axis: Association with and effect on diabetic retinopathy | |
| CN111588763A (en) | Thrombus dredging medicine, preparation method and content determination method | |
| CN111084776A (en) | Use of pilenicacid and derivatives thereof for treating and/or preventing neurodegenerative diseases | |
| Tang et al. | Cornel iridoid glycoside improves locomotor impairment and decreases spinal cord damage in rats | |
| CN116059366A (en) | Construction of nano-drug carrying p16-siRNA and application of nano-drug in treatment of post-infarction ventricular remodeling | |
| CN108420804A (en) | A kind of load Astragaloside IV pla-pcl film and its preparation method and application | |
| Meng et al. | An Innovative Model of ISS‐Based Multiple Fractures and Gastrointestinal Dysfunction Related to c‐Kit Protein Expression on Interstitial Cells of Cajal |
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 |