CN113575927B - Health-care product composition with neuroprotection and memory improving functions and preparation method thereof - Google Patents
Health-care product composition with neuroprotection and memory improving functions and preparation method thereof Download PDFInfo
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Classifications
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- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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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
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- A—HUMAN NECESSITIES
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- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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- Animal Behavior & Ethology (AREA)
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- Neurology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
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- Polymers & Plastics (AREA)
- Alternative & Traditional Medicine (AREA)
- Nutrition Science (AREA)
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- Hospice & Palliative Care (AREA)
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Abstract
The invention discloses a health care product composition with nerve protection and memory improvement and a preparation method thereof, wherein the health care product composition is prepared into a compound preparation according to the combination theory of traditional Chinese medicine compatibility theory and traditional Chinese medicine and western medicine, mixed extracts of coralloid hericium erinaceus, armillaria mellea, boletus and Morchella are taken as main raw materials, and extracts obtained after mixing in a certain proportion are taken as compound health care products, and the memory and cognitive ability are improved through synergistic effect.
Description
Technical Field
The invention belongs to the technical field of health-care product processing, in particular to a health-care product composition with the functions of protecting nerves and improving memory, and also provides a preparation method of the health-care product composition.
Background
Alzheimer's Disease (AD) results in decreased memory and other cognitive abilities, which is responsible for more than 60% of cases of dementia. The existing medicines on the market are mainly used as brain metabolism improvers or neuroprotectors aiming at single targets, but the effects are very low. In recent years, with the gradual penetration of research on pharmacological activity of traditional Chinese medicines, the clinical application of bacterial medicines to the treatment of AD patients is remarkable in auxiliary treatment effect, and the alleviation of AD symptoms and progress of the bacterial medicines show considerable development trend. Since Alzheimer's disease cannot be cured at present, prevention becomes the only choice for middle-aged and elderly people, and related functional foods become needed.
Disclosure of Invention
The invention provides a health care product composition with neuroprotection and memory improving functions, which can improve memory and cognitive ability and has obvious antioxidant and anti-aging effects.
The invention provides a health care product composition with nerve protection and memory improvement, which comprises the following raw materials in parts by weight: 40-65 parts of coralloid hericium erinaceus powder, 20-40 parts of armillaria mellea powder, 5-15 parts of boletus powder and 10-20 parts of morchella esculenta powder.
Preferably, the health-care product composition with the functions of neuroprotection and memory improvement comprises the following raw materials in parts by weight: 50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of Morchella esculenta powder.
The invention provides a preparation method of a health care product composition with nerve protection and memory improvement, which is prepared by the following steps:
(1) Weighing the raw materials according to a proportion, respectively sieving with a 50-mesh sieve, uniformly mixing, adding water, and decocting for 1.5-3 hours, wherein the mass of each time of adding water is 10-40 times of the mass of the composition, so as to obtain decoction;
(2) Filtering the decoction, concentrating under reduced pressure, and spray drying to obtain powder.
Preferably, the health-care product composition with the functions of neuroprotection and memory improvement is prepared by the following steps:
(1) Weighing the raw materials according to a proportion, respectively sieving with a 50-mesh sieve, uniformly mixing, adding water, and decocting for 2.5 hours, wherein the mass of each time of adding water is 30 times of the mass of the composition, so as to obtain decoction;
(2) Filtering the decoction, concentrating under reduced pressure, and spray drying to obtain powder.
The invention also provides application of the health-care product composition with the functions of neuroprotection and improving memory in preparation of a preparation for improving memory and cognitive ability.
The preparation for improving the memory can be powder, oral liquid, granule, tablet, capsule, pill, syrup or paste.
The preparation for improving memory and cognitive ability can be added with pharmaceutically acceptable conventional auxiliary materials.
[ usage amount ] 3.0-6.0 g/(60 kg human body);
[ applicable ] memory decline; neurasthenia and has antioxidant effect.
The pharmacological properties of the invention are as follows: the polysaccharide components are taken as main components in different fungus extracts, and as different fungus polysaccharides have different structures, polarities and molecular weights, the action targets are not necessarily the same, so that the fungus extracts have the effects of protecting and enhancing the central nervous system by combining coralloid Hericium erinaceus, delaying senescence, hypnotizing and sedating, resisting oxidation and fatigue of bolete, resisting fatigue and aging of Morchella, protecting cardiovascular system and the like, and the four fungus powders are mixed according to a certain proportion, and the extracts obtained through extraction are taken as compound health products, so that the memory and cognitive ability are improved through synergistic effect, and the fungus powder has an antioxidant effect and a better effect compared with the fungus powder singly used.
The invention has the following beneficial effects:
the compound preparation is prepared according to the combination theory of traditional Chinese medicine compatibility theory and traditional Chinese and Western medicine, mixed extracts of coralloid Hericium erinaceus, armillariella mellea, boletus and Morchella are taken as main raw materials, and the extracts obtained after mixing in a certain proportion are taken as compound health care products, so that the compound preparation can improve memory and cognitive ability through synergistic effect, has an antioxidation effect, and has better effect compared with the single use of one fungus powder.
Detailed Description
The invention is further illustrated below with reference to examples. These examples are merely illustrative and do not limit the scope of the invention in any way. It will be understood by those skilled in the art that various changes and substitutions of details and forms of the technical solution of the present invention may be made without departing from the spirit and scope of the present invention, but these changes and substitutions fall within the scope of the present invention.
The coralloid Hericium erinaceus powder used in examples and comparative examples was obtained by pulverizing fruiting bodies of coralloid Hericium erinaceus produced from Changbai mountain in a pulverizer, and was brown yellow powder for subsequent experiments.
The armillaria mellea powder used in the examples and the comparative examples is armillaria mellea mycelium obtained by submerged liquid fermentation, and the fungus powder after centrifugation and freeze-drying and crushing by a crusher is pale yellow powder and is used for subsequent experiments.
The bolete powder used in examples and comparative examples was obtained by pulverizing bolete fruiting bodies in a pulverizer, and was brown powder for use in the subsequent experiments.
The morchella powder used in the examples and the comparative examples is obtained by crushing morchella fruiting bodies by a crusher, and is light yellow powder used for subsequent experiments.
Example 1
The health care product composition with the nerve protection and memory improvement comprises the following raw material medicines:
50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of Morchella esculenta powder.
The preparation method of the composition comprises the following steps:
(1) Drying coralloid Hericium Erinaceus fruiting body, morchella mycelium and Armillariella mellea, pulverizing, and sieving with 50 mesh sieve to obtain fungus powder;
(2) 50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of morchella powder are respectively and uniformly mixed to obtain the health-care product composition with the functions of neuroprotection and memory improvement of the embodiment 1.
Example 2
The health care product composition with the nerve protection and memory improvement comprises the following raw material medicines:
40 parts of coralloid hericium erinaceus powder, 30 parts of armillaria mellea powder, 10 parts of boletus powder and 20 parts of Morchella esculenta powder.
The preparation method of the composition comprises the following steps:
(1) Drying coralloid Hericium Erinaceus fruiting body, morchella mycelium and Armillariella mellea, pulverizing, and sieving with 50 mesh sieve to obtain fungus powder;
(2) Respectively taking 40 parts of coralloid hericium erinaceus powder, 30 parts of armillaria mellea powder, 10 parts of boletus powder and 20 parts of morchella powder, adding deionized water according to a liquid-to-material ratio of 20:1, and decocting for 3 hours to obtain a decoction;
(3) Filtering the decoction, concentrating under reduced pressure, spray drying to obtain the health product composition with neuroprotection and memory improving effects of example 2, and making into various dosage forms such as granule, tablet, capsule, powder, pill, syrup or unguent according to conventional food preparation process, which is used for the following pharmacodynamic experiment.
Example 3
The health care product composition with the nerve protection and memory improvement comprises the following raw material medicines:
50 parts of coralloid hericium erinaceus powder, 20 parts of armillaria mellea powder, 10 parts of boletus powder and 20 parts of Morchella esculenta powder.
The preparation method of the composition comprises the following steps:
(1) Drying coralloid Hericium Erinaceus fruiting body, morchella mycelium and Armillariella mellea, pulverizing, and sieving with 50 mesh sieve to obtain fungus powder;
(2) Respectively taking 50 parts of coralloid hericium erinaceus powder, 20 parts of armillaria mellea powder, 10 parts of boletus powder and 20 parts of morchella powder, adding deionized water according to a liquid-to-material ratio of 25:1, and decocting for 2 hours to obtain a decoction;
(3) Filtering the decoction, concentrating under reduced pressure, spray drying to obtain the health product composition with neuroprotection and memory improving effects of example 3, and making into various dosage forms such as granule, tablet, capsule, powder, pill, syrup or unguent according to conventional food preparation process, which is used for the following pharmacodynamic experiment.
Example 4
The health care product composition with the nerve protection and memory improvement comprises the following raw material medicines:
60 parts of coralloid hericium erinaceus powder, 20 parts of armillaria mellea powder, 5 parts of boletus powder and 15 parts of Morchella esculenta powder.
The preparation method of the composition comprises the following steps:
(1) Drying coralloid Hericium Erinaceus fruiting body, morchella mycelium and Armillariella mellea, pulverizing, and sieving with 50 mesh sieve to obtain fungus powder;
(2) 60 parts of coralloid hericium erinaceus powder, 20 parts of armillaria mellea powder, 5 parts of boletus powder and 15 parts of morchella powder are respectively taken, deionized water is added according to a liquid-to-material ratio of 30:1, and the mixture is decocted for 1.5 hours to obtain decoction;
(3) Filtering the decoction, concentrating under reduced pressure, spray drying to obtain the health product composition with neuroprotection and memory improving effects of example 4, and making into various dosage forms such as granule, tablet, capsule, powder, pill, syrup or unguent according to conventional food preparation process, which is used for the following pharmacodynamic experiment.
Example 5
The health care product composition with the nerve protection and memory improvement comprises the following raw material medicines:
50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of Morchella esculenta powder.
The preparation method of the composition comprises the following steps:
(1) Drying coralloid Hericium Erinaceus fruiting body, morchella mycelium and Armillariella mellea, pulverizing, and sieving with 50 mesh sieve to obtain fungus powder;
(2) Respectively taking 50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of morchella powder, adding deionized water according to a liquid-to-material ratio of 30:1, and decocting for 2.5 hours to obtain decoction;
(3) Filtering the decoction, concentrating under reduced pressure, spray drying to obtain the health product composition with neuroprotection and memory improving effects of example 5, and making into various dosage forms such as granule, tablet, capsule, powder, pill, syrup or unguent according to conventional food preparation process, which is used for the following pharmacodynamic experiment.
Comparative example 1
The comparative example is Hericium coralloides fungus powder.
The preparation method of the comparative example comprises the following steps: drying fruiting body of Hericium coralloides, pulverizing with pulverizer, and sieving with 50 mesh sieve to obtain Hericium coralloides powder.
Comparative example 2
The comparative example is Armillariella mellea powder.
The preparation method of the comparative example comprises the following steps: freeze-drying the armillaria mellea mycelium obtained by fermentation, crushing by a crusher, and sieving by a 50-mesh sieve to obtain armillaria mellea powder.
Comparative example 3
The comparative example is bolete powder.
The preparation method of the comparative example comprises the following steps: drying the bolete fruiting body, pulverizing with pulverizer, and sieving with 50 mesh sieve to obtain bolete powder.
Comparative example 4
The comparative example is Morchella esculenta powder.
The preparation method of the comparative example comprises the following steps: drying Morchella fruit body, pulverizing with pulverizer, and sieving with 50 mesh sieve to obtain Morchella powder.
Test example 1
The test example is to detect the neuroprotective effect of the health care product composition with neuroprotection and memory improvement.
(1) Test materials
The test drugs were the healthcare product compositions with neuroprotection and memory improvement obtained in examples 1, 2, 3, 4 and 5 and the coralloid Hericium erinaceus powder, armillariella mellea powder, boletus powder and Morchella powder of comparative examples 1, 2, 3 and 4. The mouse cell line used in this experiment was a mouse primordial hippocampal neuronal cell line (HT 22; BNCC; 337709) purchased from North Naraxacum Biotechnology.
(2) Test method
The neuroprotection of individual bacterial compositions of the above examples and comparative examples against L-glutamate induced HT22 cell damage was examined. HT22 cells at 5X 10 3 Cell concentration of each was seeded in 96-well plates at 5% CO 2 Cells were pretreated with doses of 25 and 100. Mu.g/mL of each group of drugs for 3 hours after observing the cell status in a 37℃incubator culture of about 14 h, then modeled with L-glutamic acid (L-Glu) at a final concentration of 25 mM, and the blank group was not treated, while again being administered as a pretreatment, and incubated for 24 hours. 5 mg/mL MTT solution, 10 mu L/well, was added to the 96-well plate in the dark, incubated for 4 hours in the dark, the precipitate was dissolved in DMSO, absorbance was measured at 490/540 nm, and the change in cell viability was analyzed.
The survival rate of each group was calculated with the survival rate of the blank group being 100%. Experimental data are expressed as mean ± standard deviation (mean ± s.d.). Statistically significant differences were tested for group differences (post hoc Dunn's test) according to one-way ANOVA analysis of IBM SPSS Statistics software, P <0.05 considered the group differences significant, with statistical significance.
(3) Test results
The results in table 1 show the effect of single components of the health care product composition with neuroprotection and memory improvement obtained in examples 1, 2, 3, 4 and 5 and the coralloid Hericium erinaceus powder, armillaria mellea powder, boletus powder and Morchella esculenta powder of comparative examples 1, 2, 3 and 4 on the viability of HT22 cells, respectively. Compared with the blank, the cell viability of the control group is reduced to 51.7% (P < 0.01) after modeling by L-glutamic acid (final concentration of 25 mM), the four bacterial powders of the control group all show no obvious neuroprotection when the administration concentration is 25 mug/mL, the survival rate of cells is obviously improved (P < 0.05) when the dosage is increased to 100 mug/mL, except for boletus powder, hericium coralloides, armillariella mellea and Morchella, the neuroprotection effect of the example 1 obtained by mixing the four bacterial powders in a certain proportion on HT22 cells is better than that of the single component, and the bacterial composition obtained by leaching the comparative examples 2, 3, 4 and 5 by hot water has obvious neuroprotection (P < 0.05) when the administration concentration is 25 mug/mL, wherein the effect of the example 5 is the best, and the cell viability is improved to 86.5% (P < 0.01). The composition has the advantages that the pretreatment and the administration of the health-care product composition with the functions of neuroprotection and memory improvement have obvious neuroprotection effect on L-glutamic acid-induced cell injury, and compared with comparative examples 1-4, the composition extract has obvious neuroprotection effect compared with single-component fungus powder, and the composition has better effect after compatibility.
TABLE 1 neuroprotection and memory enhancing results of health care compositions on HT22 cells
| Cell viability/% | Drug administration concentration/(25. Mu.g/mL) | P value | Drug administration concentration/(100. Mu.g/mL) | P value |
| Blank group | 100.0±3.5 | --- | 100.0±3.5 | --- |
| ControlGroup of | 51.7±7.6 | P<0.01 | 51.7±7.6 | P<0.01 |
| Example 1 | 54.2±3.6 | P>0.05 | 62.8±6.7 | P>0.05 |
| Example 2 | 58.5±2.4 | P>0.05 | 64.0±4.4 | P<0.05 |
| Example 3 | 64.8±5.9 | P<0.01 | 73.5±3.9 | P<0.01 |
| Example 4 | 63.0±7.3 | P<0.05 | 75.5±4.5 | P<0.01 |
| Example 5 | 76.2±11.2 | P<0.01 | 86.5±6.2 | P<0.01 |
| Comparative example 1 | 53.5±2.7 | P>0.05 | 61.3±4.6 | P<0.01 |
| Comparative example 2 | 49.2±3.8 | P>0.05 | 60.3±5.4 | P<0.05 |
| Comparative example 3 | 52.0±2.8 | P>0.05 | 54.0±5.5 | P>0.05 |
| Comparative example 4 | 51.5±6.3 | P>0.05 | 62.7±5.0 | P<0.05 |
Note that: p >0.05 represents no significance, p <0.05 represents significant, and p <0.01 represents very significant.
Test example 2
The test example is to detect the influence of a health care product composition with neuroprotection and memory improvement on the memory improvement and antioxidation capability of mice.
(1) Test materials
Medicament: physiological saline, the health care product composition with neuroprotection and memory improvement obtained in examples 1, 2, 3, 4 and 5, and coralloid Hericium erinaceus powder, armillariella mellea powder, boletus powder and Morchella powder of comparative examples 1, 2, 3 and 4.
Animals: 60 SPF-grade C57BL/6 male mice (8 weeks old, body weight 18-22 g) were purchased from the vinca biologicals institute, license number: SCXK (gei) -2011-0003. All mice were kept in an environment with a temperature of 23.+ -. 1 ℃ and a humidity of about 55%. The lighting time is kept for 12 hours, natural light is evenly irradiated, and 6 mice/cage can freely ingest food and water.
The kit comprises: biochemical index detection includes ELISA kits of central cholinergic related indexes such as acetylcholine (ACh), acetylcholinesterase (AChE), choline acetyltransferase (ChAT), oxidative stress related indexes including ROS, superoxide dismutase (SOD), all of which are purchased from Jiangsu kott biotechnology, inc, all of which are operated according to instructions provided in the kit.
(2) Test method
Mice were grouped and dosed: SPF-class healthy C57BL/6 male mice are selected to be 60, randomly divided into 10 groups of 6 mice each, and the groups are respectively as follows: blank, example 1, example 2, example 3, example 4, example 5, comparative example 1 (Hericium coralloides powder), comparative example 2 (Armillariella mellea powder), comparative example 3 (Boletus powder) and comparative example 4 (Morchella powder) for 10 total. Each group was dissolved with distilled water to the appropriate drug concentration. The administration was performed by gavage at 9:00 a.m. each day at a dose of 10 mg/kg and a gavage volume of 0.1mL/20g body weight, and the blank group was gavaged with physiological saline for 3 weeks.
Animal behavioural test: the effect of the health-care product composition with neuroprotection and memory improvement on the cognition and memory improvement of mice is examined through Morris water maze.
The water maze consists of a stainless steel pool (120 cm a diameter, 50 a height cm a) and an immersed escape platform (10 a cm a diameter) 1cm below the water surface. Tap water is injected into the water maze to enable the tap water to pass through the escape platform 1 and cm, titanium dioxide powder is added into the water, and the water is stirred uniformly to enable the water to be white and opaque, and the water temperature is kept at about 25 ℃. The water maze is equally divided into 4 1/4 circles by the water maze operation system and is defined as four quadrants. The escape platform is placed in the second quadrant, and the mice are put into water from the same position each time. Each experiment ensures that the environment is quiet. The time required for the mice to find the hidden platform was recorded as escape latency for 4 consecutive days of spatial learning testing. The experimental time is set to be 2min, and the mice which do not find the platform within the set time are artificially guided to the escape platform. The time for the mice to find the platform was recorded by the water maze monitoring system on day 5.
After the last behavioral test, mice were fasted for 12 hours. Blood was sampled from the tail vein, allowed to stand at room temperature for 30min, centrifuged at 3500rpm for 5 minutes, and after centrifugation, the supernatant was collected and stored in a-80 ℃ refrigerator. Mice were then euthanized by intraperitoneal injection of sodium pentobarbital (100 mg/kg). Brain tissue was collected rapidly.
Central cholinergic related index determination in the hippocampus of mice: the collected hippocampal tissues were homogenized with physiological saline and the protein concentration was detected by BCA kit (Millipore). The assay was performed according to the instructions provided for the kit to detect AChE, ach, chAT levels in the hippocampus of mice.
Measurement of oxidative stress related index in mouse serum: the collected mouse serum is tested according to the instruction provided by the kit, and the SOD and ROS content in the mouse serum is detected.
The data processing method comprises the following steps: experimental data are expressed as mean ± standard deviation (mean ± s.d.). Statistically significant differences were tested for group differences (post hoc Dunn's test) according to one-way ANOVA analysis of IBM SPSS Statistics software, P <0.05 considered the group differences significant, with statistical significance.
(3) Experimental results
Morris water maze test results: the experimental results of the platform time of each group of mice, namely escape latency are shown in Table 2, example 1 is mixed powder of four bacteria of Hericium coralloides, armillariella mellea, boletus and Morchella, the platform time of the mice can be obviously shortened after 3 weeks of administration (P < 0.05), and the experimental results are superior to those of the single component administration, namely comparative examples 2-4 (P > 0.05), and the effect of the single component administration is similar to that of the coralloid Hericium coralloides powder of comparative example 1. The four bacterial powders in examples 2-5 are mixed in a certain proportion and then subjected to hot water leaching, so that the obtained healthcare product composition with the neuroprotection and memory improvement can remarkably shorten the time for searching the platform of the mice (P < 0.05), the time for searching the platform of the mice can be remarkably shortened in examples 3 and 5, and the spatial memory of the mice is improved, wherein the effect of example 5 is most obvious, and compared with mice in a blank control group, the time is shortened by 39.6% (P < 0.01). This demonstrates a significant improvement in cognitive and memory in mice following administration of the compositions of examples 1-5. In addition, as a result of comparing the examples with the comparative examples in the lateral direction, the effect of reducing the mouse seeking time due to the synergistic effect of the examples is more obvious, significance (P <0.05 and P < 0.01) is universally present, and the effect of the comparative examples is mostly insignificant (P > 0.05).
TABLE 2 influence of health care composition with neuroprotection and memory enhancement on mouse Water maze experiments
| Group of | Number of animals/animal | Escape latency/s | P value |
| Blank control group | 6 | 45.7±5.9 | |
| Example 1 | 6 | 37.5±5.4 | P<0.05 |
| Example 2 | 6 | 36.2±7.1 | P<0.05 |
| Example 3 | 6 | 38.0±3.7 | P<0.01 |
| Example 4 | 6 | 38.8±4.4 | P<0.05 |
| Example 5 | 6 | 31.3±4.8 | P<0.01 |
| Comparative example 1 | 6 | 37.7±5.8 | P<0.05 |
| Comparative example 2 | 6 | 39.8±4.8 | P>0.05 |
| Comparative example 3 | 6 | 39.5±4.4 | P>0.05 |
| Comparative example 4 | 6 | 40.3±5.2 | P>0.05 |
Note that: data are expressed as mean ± standard deviation. (n=6/group). P >0.05 represents no significance, P <0.05 represents significance, and P <0.01 represents extreme significance.
Central cholinergic related index determination results: acetylcholine (Ach) associated with learning memory and cognitive abilities is an important neurotransmitter in the brain reflecting the status of cholinergic neurons. Neurotransmitter Ach acts on many cholinergic receptors and is also an excitatory neurotransmitter of many motor nerves, and an increase in its content is associated with an improvement in alzheimer's disease. Both AchE as Ach hydrolase and ChAT as Ach synthase can be used to regulate Ach metabolism.
As shown in table 3, the results of experiments show that, after the mice were subjected to gastric administration for 3 weeks, the single fungus powder of comparative examples 1 to 4 was administered only to coralloid Hericium erinaceus to significantly reduce the level of AchE (P < 0.05), the levels of AchE in the hippocampus of the mice were not significantly changed by armillaria mellea, boletus and Morchella (P > 0.05), and the levels of AchE were significantly reduced in examples 2 to 5 obtained by hot water extraction of the four fungus mixtures mixed in a certain ratio. Examples 1-5 significantly increased the Ach content in the hippocampus of mice (P < 0.05) compared to the placebo group, with example 5 being optimally effective at 1.23 times the placebo group, whereas the administration of a single powder intragastric gavage in the comparative example did not significantly change the Ach content in the hippocampus of mice (P > 0.05). For ChAT, example 5 significantly increased ChAT levels in the hippocampus of mice (P < 0.05) compared to the placebo group, while examples 1-4 all significantly increased ChAT levels (P < 0.01), with example 3 being most effective. The experimental results of comparative examples 1 and 3 show that the single powder administration of armillaria mellea and morchella also significantly increases the ChAT level (P < 0.05) in the hippocampus of mice, but the effect is not significant in the examples. By combining AChE, ach and ChAT experimental results, it can be seen that the health care product composition with neuroprotection and memory improvement can achieve neuroprotection by improving the level of central cholinergic related indexes, wherein the level of Ach and ChAT can be obviously changed in example 1 prepared by mixing four bacterial powders in a certain proportion. The health-care product composition with the nerve protection and memory improvement obtained after hot water extraction shows better effect. The comparative examples have the same effects, but the effects of the individual effects are not obvious and are not remarkable because the action routes of the different bacteria are different.
TABLE 3 effects of health care compositions with neuroprotection and memory enhancement on Central cholinergic related indicators in mice
| Group of | Number of animals/animal | AChE/（U/mg prot） | Ach/（μg/ mg prot） | ChAT/（pg/ mg prot） |
| Blank control group | 6 | 1.3±0.2 | 367.7±21.4 | 150.8±14.4 |
| Example 1 | 6 | 1.0±0.2 | 394.0±16.9* | 175.2±10.0** |
| Example 2 | 6 | 1.0±0.0** | 398.2±14.1* | 181.7±12.8** |
| Example 3 | 6 | 0.9±0.1** | 392.0±15.0* | 201.8±11.7** |
| Example 4 | 6 | 0.8±0.1** | 404.6±22.1* | 189.7±8.5** |
| Example 5 | 6 | 0.8±0.1** | 451.7±75.2* | 175.2±19.3* |
| Comparative example 1 | 6 | 1.0±0.1* | 385.1±19.3 | 150.5±16.0 |
| Comparative example 2 | 6 | 1.1±0.2 | 362.6±35.8 | 173.2±17.2* |
| Comparative example 3 | 6 | 1.0±0.2 | 376.8±56.2 | 163.3±17.9 |
| Comparative example 4 | 6 | 1.2±0.2 | 381.2±49.2 | 177.7±22.7* |
Note that: in comparison with the blank group, * P<0.05， ** P<0.01。
oxidative stress related index measurement results: oxidative stress is the basis of pathophysiological important assumptions for many diseases, including neurodegenerative diseases. The SOD serving as natural enemy of oxygen free radicals is scavenger of organisms, can remove harmful substances generated in the organisms, has an anti-aging effect, can prevent the generation of the oxygen free radicals, and can prevent the generation of the oxygen free radicals from damaging cells, thereby repairing the damaged cells. The experimental results are shown in table 4, and compared with the blank control group, the SOD content in the serum of the mice can be obviously improved (P < 0.05) in the example 2, and meanwhile, the SOD content in the serum of the mice can be obviously improved (P < 0.01) in the examples 3, 4 and 5. In addition, the SOD content in the serum of the mice is also improved in example 1 and comparative example, but the effect is not significant. In addition, examples 2, 4 and 5 and comparative example 3 can significantly scavenge ROS in mouse serum, on the basis of which the scavenging effect of examples 2 and 3 on ROS is extremely pronounced (p < 0.01), wherein the ROS level of example 2 is reduced by 33.6% (p < 0.01) compared to the blank. In conclusion, the health care product composition with the function of neuroprotection and memory improvement can improve the oxidative stress state of mice. Compared with the single-component bacterial powder of comparative examples 1-4, the four bacterial powders compounded in a certain proportion show better antioxidation effect after hot water leaching.
TABLE 4 influence of compositions with neuroprotection and memory enhancing effects on oxidative stress index levels in mouse serum
| Group of | Number of animals/animal | SOD/（U/ mg prot） | ROS/（U/ mg prot） |
| Blank control group | 6 | 147.2±17.9 | 53.0±11.5 |
| Example 1 | 6 | 164.7±14.0 | 40.5±3.8* |
| Example 2 | 6 | 173.5±13.9* | 35.2±2.3** |
| Example 3 | 6 | 181.5±8.1** | 36.2±3.7** |
| Example 4 | 6 | 211.8±24.1** | 39.0±3.7* |
| Example 5 | 6 | 205.5±25.6** | 37.2±4.5* |
| Comparative example 1 | 6 | 157.5±19.2 | 44.5±2.3 |
| Comparative example 2 | 6 | 165.3±16.7 | 46.8±4.8 |
| Comparative example 3 | 6 | 155.7±21.2 | 40.0±3.3* |
| Comparative example 4 | 6 | 155.0±18.0 | 45.2±3.3 |
Note that: in comparison with the blank group, * P<0.05， ** P<0.01
in vitro cell experiments show that the health-care product composition with neuroprotection and memory improvement obtained after hot water leaching has better neuroprotection effect than single fungus powder, and the same experimental result is verified in animal experiments. The experimental results show that each example shows good memory and cognition improving ability and has obvious antioxidation and anti-aging effects by comparing with the blank control group and the comparative example, namely the single-component bacterial powder. Example 1 is an untreated mixed bacterial powder, which shows a certain memory improving effect, but examples 2-5 obtained after hot water leaching show more excellent effect, and in conclusion, the composition of the invention can be used as a health care product composition for improving cognition and memory.
Claims (1)
1. A health-care product composition is characterized by being prepared from the following raw materials in parts by weight:
50 parts of coralloid hericium erinaceus powder, 25 parts of armillaria mellea powder, 10 parts of boletus powder and 15 parts of Morchella esculenta powder;
the coralloid Hericium erinaceus powder is obtained by pulverizing fruiting body of coralloid Hericium erinaceus with pulverizer;
the armillaria mellea powder is armillaria mellea mycelium obtained by submerged liquid fermentation, and is obtained by crushing the armillaria mellea mycelium by a crusher after centrifugation and freeze-drying;
the boletus bacteria powder is obtained by crushing boletus fruiting bodies through a crusher;
the Morchella powder is obtained by pulverizing Morchella fruiting body with a pulverizer;
the preparation method of the health-care product composition comprises the following steps:
1) Weighing the raw materials according to a proportion, respectively sieving with a 50-mesh sieve, uniformly mixing, adding water, and decocting for 2.5 hours, wherein the mass of each time of adding water is 30 times of the mass of the composition, so as to obtain decoction;
2) Filtering the decoction, concentrating under reduced pressure, and spray drying to obtain powder.
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