CN114796179A - Application of myristic acid in preparation of anti-brain-aging drugs - Google Patents
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
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- 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
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Abstract
The invention provides an application of myristic acid in preparation of a brain aging resistant medicine. The invention discovers that myristic acid can effectively improve the exploration capacity and the spatial memory of a naturally aging mouse, improve the degeneration of hippocampus neurons, increase the percentage of hippocampus nissl positive cells and reduce hippocampus aging marker proteins P16 and P21. Thus, myristic acid is effective against brain aging, and can be used for treating brain aging and brain aging-related diseases, including Alzheimer's disease and brain atrophy.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of myristic acid in preparation of a brain aging resistant medicine.
Background
With the rapid increase of the global aging population, brain aging and related diseases (such as alzheimer's disease, brain atrophy, etc.) place a huge burden on patients, families, and socioeconomic. Alzheimer's Disease (AD), also known as alzheimer's disease, dementia of the elderly, senile dementia, cerebral degenerative disease, is a persistent neurological dysfunction, which is also the most common cause of dementia, and the symptoms are manifested as gradually severe cognitive disorders (memory disorders, learning disorders, attention disorders, spatial cognitive functions, disorders of problem solving ability), gradually failing to adapt to society. At present, no medicine for specifically reversing brain aging exists, so that the search for a therapeutic medicine for brain aging is imminent. The hippocampus is an important component of the limbic system of the brain and contains various messenger receptors, which are not only the higher regulatory centres of the neuroendocrine system, but also the most responsive areas. Research proves that the hippocampus function is closely related to aging, and the hippocampus neurons are reduced along with the aging of the organism. Therefore, a more thorough understanding of hippocampus aging is crucial for improving the treatment of brain aging-related diseases.
Myristic acid (hereinafter referred to as MA, IUPAC name: myristic acid, molecular formula: CH3(CH2)12COOH, CAS #: 544-63-8) is a saturated fatty acid, mainly present in cardamom oil, coconut oil and palm kernel oil. The effects of MA on hippocampal neurons and brain aging have not been reported.
Disclosure of Invention
Based on the above, the invention aims to provide the application of myristic acid in preparing a medicament for resisting brain aging. Myristic acid is effective in improving the exploration ability, spatial memory and hippocampus aging of naturally aging mice.
In order to achieve the purpose, the invention adopts the following technical scheme.
Application of myristic acid in preparing medicine for resisting brain aging is provided.
In some embodiments, the drug may improve spatial memory.
In some embodiments, the drug may enhance exploratory ability.
In some embodiments, the brain aging is hippocampus aging.
In some embodiments, the medicament may improve hippocampal neuronal degeneration.
In some embodiments, the drug can increase the number of niemann-positive cells in the hippocampus.
In some embodiments, the drug may reduce the expression of p16Ink4a in hippocampus.
The invention also provides application of myristic acid in preparation of a medicine for treating Alzheimer disease.
The invention also provides application of myristic acid in preparation of a medicine for treating brain atrophy.
The invention also provides a medicament for resisting brain aging, which comprises myristic acid and a pharmaceutically acceptable carrier.
The invention discovers that myristic acid can effectively improve the exploration capacity and the spatial memory of a naturally aging mouse, improve the degeneration of hippocampus neurons, increase the percentage of hippocampus nissl positive cells and reduce hippocampus aging marker proteins P16 and P21. Thus, myristic acid is effective against brain aging, and can be used for treating brain aging and brain aging-related diseases, including Alzheimer's disease and brain atrophy.
Drawings
FIG. 1 shows the results of the evaluation of the exploratory ability and spatial memory ability of MA-treated naturally aging mice; wherein, A: naturally aging mice (22M) and MA intervention group (MA +22M) open field test parameters, P < 0.05; b: a Morris water maze track diagram from 1d to 4d, and a positioning navigation test; 5d, space exploration test; c: morris water maze escape latency,. P <0.01 and. P < 0.001; d: number of platform crossings by Morris water maze experiment,. P < 0.05.
FIG. 2 shows the result of MA treatment for the evaluation of the morphology of the hippocampus and the aging index of the spontaneously aging mice; wherein, A: h & E and niemann staining of hippocampus; B-C: number of hippocampus ni positive cells and neuronal degeneration rate, # P < 0.01; d: IHC shows an under-mirror image of hippocampus senescence markers P16 and P21; E-F: relative quantification of IHC for aging markers P16 and P21, × P < 0.01.
Detailed Description
Experimental procedures according to the invention, in which no particular conditions are specified in the following examples, are generally carried out under conventional conditions, or under conditions recommended by the manufacturer. The various chemicals used in the examples are commercially available.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, article, or apparatus that comprises a list of steps is not limited to only those steps or modules recited, but may alternatively include other steps not recited, or may alternatively include other steps inherent to such process, method, article, or apparatus.
The following description will be given with reference to specific examples.
Example 1
This example uses a naturally aging mouse to perform an experiment to investigate the therapeutic effect of MA on brain aging.
Experimental materials and methods
1. Mouse
C57BL/6 male mice were purchased at 22 months of age from the university of traditional Chinese medicine laboratory animal center (GZUCM) (animal production certificate # (Yue)20180034) in guangzhou. All animal experiments were approved by the ethical committee of the first subsidiary hospital of the university of traditional Chinese medicine in Guangzhou (approval document TCMF 1-2021026). 22-month-old mice were randomly divided into two groups: 22-month-old mice (22M, i.p. with equal amounts of solvent) and MA (MA +22M, i.p. with 2 mg/kg. d MA). The medicine is administered by intraperitoneal injection for 2 months. Following behavioral assessments (open field and water maze experiments), mouse brain samples were taken for subsequent experimental assessments.
2. Open field experiment
Open field experiments (for evaluating mouse exploration behavior) the procedure was as follows: mice were transported 2h in advance to the laboratory to adapt to the pre-experimental environment. The reaction chamber of the C57BL/6 mouse in the open field experiment is 30cm high, the bottom surface is 72cm long, and the inner wall is white. The bottom of the reaction chamber was wiped with 75% alcohol in open field experiments. The animals were gently removed from their cages and quickly placed in the open field laboratory reaction box. Parameters are set in SuperMaze software (Shanghai Xin Soft information technology Co., Ltd., version 3.0), and the activity of a mouse in an open field experiment reaction box is automatically recorded for 5 min. After the first mouse completed the experiment, the bottom of the box was wiped with 75% alcohol to remove the odor, and the next mouse was tested in the same manner as before. The parameters include: total Distance of movement of the mouse (Total Distance), Time of rest of the mouse (Quiescent Time), Distance of movement of the central zone (Distance Center), Number of entries of the central zone (Number of Center), residence Time of the central zone (Time Center), Distance of movement of the peripheral zone (Distance perimeter), Number of entries of the peripheral zone (Number of perimeter), residence Time of the peripheral zone (Time perimeter), residence Time of the central zone/residence Time of the peripheral zone (Time Center/Time perimeter).
Morris Water maze
The Morris water maze was used to assess spatial memory in mice. After the open field experiment, the Morris water maze test was performed. The mice are placed into the pool from four quadrants facing the pool wall, and the time required from entering the water to standing on an underwater hidden platform is recorded as the escape latency by SuperMaze software (Shanghai Xin Soft information technology Co., Ltd., version 3.0). If the mouse did not find the platform after 60s into the water, the mouse was guided to stay on the platform for 10 s. Four quadrants per day for a total of four days. On the fifth day, a space exploration experiment was performed. The water maze platform was removed and the mice were placed into the water from a position opposite the platform. The swimming time of the mice is 60 s. The quadrants trained each day are randomly arranged. Recording parameters: escape Latency (Escape Latency) and number of platform crossings (Platformcrossover number).
4. Hippocampus HE staining and Nie staining
The cerebral hemispheres were removed intact on ice, fixed with 4% paraformaldehyde for 48h, and then embedded in paraffin. The hemispheres were dissected in the midsagittal plane to the hippocampal region at a slice thickness of 5 μm and stained with H & E staining solution (Sericebio, G1005, Wuhan) and Nissl staining solution (Sericebio, G1036, Wuhan), respectively. Hippocampus slices H & E and Nissl staining were intelligently taken by a digital pathology scanning system (3D HISTEC ltd., panoramic MIDI).
5. Immunohistochemical staining (IHC)
IHC was used to detect the expression of hippocampus P16 and P21 proteins. Brain tissue slices are sequentially deparaffinized and antigen repaired. Primary anti-P16 (Affinity Biosciences Cat # AF0228, RRID: AB _2833403, Rabbit,1:200) and P21(Beyotime Cat # AP021,1:200) antibodies were added and incubated overnight at 4 ℃. Biotin-labeled secondary antibody was then added and incubated for 1h at room temperature. After conventional DAB staining, hematoxylin was stained and mounted. Digital pathology scanning system (3D HISTEC ltd., panoramic MIDI) smart photography. Image J1.51 analytical software (Wayne Rasband, national institute of health, usa) was used to calculate relative protein expression.
6. Statistical analysis
All data were checked for normality and homogeneity of variance. If two independent sample data were normally distributed, the results were expressed as mean ± Standard Deviation (SD) and a two-sided independent sample t-test was used. If the two independent sample data do not fit normally, the Mann-Whitney U test is used and a box plot is used. Morris water maze data were analyzed using two-way analysis of variance (ANOVA) in combination with Bonferroni multiple comparisons. P <0.05 was considered to be statistically different. Mapping and statistical analysis were performed using GraphPad Prism version 8.4.3 (GraphPad Software, San Diego, California, USA).
Second, experimental results
MA improving exploration ability and spatial memory ability of naturally aging mice
Results as shown in fig. 1, we observed an increase in the exploratory capacity of MA-treated mice in the open field experiments, including an increase in Total Distance of movement (Total Distance) of the mice, Number of central zone entries (Number of Center), central zone entry Time/peripheral zone entry Time (Time Center/Time period). Reduce the resting Time (Quiescent Time) of mice (fig. 1A). In the Morris water maze, we observed a decrease in Escape Latency (Escape Latency) and an increase in number of Platform crossings (Platform cross number) in mice, indicating that MA improved hippocampus-dependent spatial memory (fig. 1B-D).
MA improvement of mouse hippocampus neuronal degeneration
H & E and niemann staining were further used to demonstrate the effect of MA on hippocampus histomorphology. As shown in figure 2, the disordered hippocampus neurons were improved in aged mice after MA treatment (figure 2A). Quantitative results indicated that the percentage of nissl positive cells increased and the neurodegeneration rate decreased (fig. 2B-C). IHC showed a significant decrease in the classical senescence marker P16, with a trend toward a decrease in P21 after MA intervention (fig. 2D-F).
In summary, MA can effectively treat brain aging, especially improve hippocampus aging, and can be used for preparing medicines for treating brain aging and brain aging related diseases.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, however, as long as there is no contradiction between the combinations of the technical features, the scope of the present description should be considered as being described in the present specification.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. Application of myristic acid in preparing medicine for resisting brain aging is provided.
2. The use of claim 1, wherein the medicament improves spatial memory.
3. The use of claim 1, wherein the medicament is capable of enhancing exploratory power.
4. The use of claim 1, wherein the brain aging is hippocampus aging.
5. The use of claim 4, wherein the medicament ameliorates hippocampal neuronal degeneration.
6. The use of claim 4, wherein the medicament is capable of increasing the number of Neisseria positive cells in the hippocampus.
7. The use of claim 4, wherein the medicament is capable of reducing the expression of P16in the hippocampus.
8. Application of myristic acid in preparing medicine for treating Alzheimer disease is provided.
9. Application of myristic acid in preparing medicine for treating brain atrophy is provided.
10. An anti-brain-aging medicament, comprising myristic acid and a pharmaceutically acceptable carrier.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102151268A (en) * | 2011-03-28 | 2011-08-17 | 王义明 | Compound preparation for treating Alzheimer's disease and preparation method thereof |
| DE202022100164U1 (en) * | 2022-01-12 | 2022-02-24 | Charan Chabbanahalli | Herbal composition with anti-amnesia and cerebro-protective potentials |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102151268A (en) * | 2011-03-28 | 2011-08-17 | 王义明 | Compound preparation for treating Alzheimer's disease and preparation method thereof |
| DE202022100164U1 (en) * | 2022-01-12 | 2022-02-24 | Charan Chabbanahalli | Herbal composition with anti-amnesia and cerebro-protective potentials |
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| Title |
|---|
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| HABIB NAZEM等: ""Chitosan‑myristate nanogel as an artificial chaperone protects neuroserpin from misfolding"", 《ADVANCED BIOMEDICAL RESEARCH》, no. 5, pages 1 - 6 * |
| MAAZ AHMED KHAN等: "Development of Synergy-Based Combination for Learning and Memory Using in vitro, in vivo and TLC-MS-Bioautographic Studies", FRONTIERS IN PHARMACOLOGY, vol. 12, 31 July 2021 (2021-07-31), pages 1 - 12 * |
| TO DAO CUONG等: "Potent Acetylcholinesterase Inhibitory Compounds from Myristica fragrans", NATURAL PRODUCT COMMUNICATIONS, vol. 9, no. 4, 30 April 2014 (2014-04-30), pages 499 - 502 * |
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