CN112535289A - NMN-containing composition capable of delaying senescence and preparation method and application thereof - Google Patents
NMN-containing composition capable of delaying senescence and preparation method and application thereof Download PDFInfo
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- CN112535289A CN112535289A CN202011493479.9A CN202011493479A CN112535289A CN 112535289 A CN112535289 A CN 112535289A CN 202011493479 A CN202011493479 A CN 202011493479A CN 112535289 A CN112535289 A CN 112535289A
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Abstract
The invention discloses a composition containing NMN and capable of delaying senescence, a preparation method and an application thereof.
Description
Technical Field
The invention relates to the technical field of food health care, in particular to a composition containing NMN and capable of delaying senescence, and a preparation method and application thereof.
Background
Aging, also called aging, is an unavoidable stage in life, is a spontaneous inevitable process of all multicellular organisms over time, has harmful changes at all levels of organisms and tissues, shows a decline process of functions, adaptability and resistance, and can be divided into physiological aging and pathological aging, wherein the physiological aging refers to gradual irreversible degenerative change of a complex morphological structure and physiological functions of the whole body, influenced by genetic factors, of an organism from the beginning of the maturation stage and along with the aging, also called normal aging. Pathological aging refers to accelerated aging caused by diseases or abnormal factors, also called abnormal aging. Aging is closely related to senile diseases, and the more serious the aging degree is, the greater the probability of the disease is. Aging is a continuous dynamic and slowly progressive process, which gradually starts from the end of the growth period, and the influence of the aging is gradually shown by the dysfunction, the decline of organ functions, the cell degeneration and the change of protein and enzyme activity of a human body system in the old period, mainly showing that the adaptive capacity of the organism to the environmental stimulation is weakened or even lost, and the decline of various tissue and organ functions appears; with the continuous development of medical level and scientific technology, human beings have conducted many studies on aging mechanism and life-prolonging and anti-aging method, and there is an urgent need to develop better dietary or pharmaceutical intervention measures for aging and aging-related disease mechanism.
beta-Nicotinamide Mononucleotide (NMN) is used as one of important precursors of mammal NAD +, and the in vivo generation reaction depends on the catalytic action of nicotinamide ribotransferase phosphate compounds, wherein the NMN in human bodies is mainly derived from foreign food intake and self-synthesis of organisms; research finds that artificial supplementation has a plurality of advantages, on one hand, in peripheral tissues and organs, the endocrine of the body is regulated, and the retinal degenerative disease is treated; improving and repairing islet sensitivity, promoting insulin secretion, and preventing and controlling metabolic diseases such as type II diabetes and obesity; protecting heart and kidney, treating cardiovascular diseases and acute renal injury, and treating brain injury and apoplexy in central nervous system, and repairing mitochondrial respiratory function deficiency of brain. In fact, one of the hallmarks of aging is the development of neurodegenerative diseases. Our common neurodegenerative diseases such as alzheimer's disease, parkinson's disease, huntington's disease, amyotrophic lateral sclerosis and frontotemporal dementia have increased susceptibility with age. Research shows that the artificial NMN supplementation can play a certain saving role in AD, PD and other neurodegenerative diseases. More importantly, NMN has non-toxic characteristics. The results show that NMN has important potential as an effective anti-aging intervention means.
In the prior art, most of products with anti-aging efficacy adopt a traditional Chinese medicine formula, but the traditional Chinese medicine formula has complex components, the efficacy and the toxic and side effects are easy to question, and although other anti-aging products on the market can enhance the function of a human body to a certain extent, the using effect is not ideal, and the anti-aging effect needs to be enhanced.
Disclosure of Invention
The invention aims to provide a composition containing NMN and capable of delaying senescence, a preparation method and application thereof, and solves the problems that anti-senescence products in the prior art are poor in effect, toxic and side effects exist and the like.
The technical scheme of the invention is as follows:
the NMN-containing composition capable of delaying senescence comprises main materials and auxiliary materials, wherein the main materials of the composition comprise: NMN, crab apple extract, pumpkin seed extract, wall-broken pine pollen, grape seed extract and blueberry extract.
Preferably, the main materials of the composition comprise the following raw materials in parts by weight: NMN1-10 parts, crab apple extract 10-20 parts, pumpkin seed extract 10-20 parts, wall-broken pine pollen 2-5 parts, grape seed extract sugar 10-15 parts, and blueberry extract 10-15 parts.
Preferably, the main materials of the composition comprise the following raw materials in parts by weight: 5 parts of NMN, 15 parts of crab apple extract, 15 parts of pumpkin seed extract, 3 parts of wall-broken pine pollen, 12 parts of grape seed extract sugar and 12 parts of blueberry extract.
Preferably, the preparation method of the malus micromalus makino extract, the pumpkin seed extract, the grape seed extract and the blueberry extract comprises the following steps:
the preparation method of the malus micromalus makino extract comprises the following steps: cleaning fresh Malus micromalus makino, naturally drying in the shade, removing core, collecting pulp, drying the collected fruit residue at low temperature and constant temperature, pulverizing, sieving with 80-100 mesh sieve, mixing Malus micromalus makino powder with 40% volume fraction ethanol solution according to a solid-to-liquid ratio of 1:35, performing microwave treatment at microwave frequency of 2450MHz and power of 550W for 180-240s, filtering to obtain extractive solution, rotary evaporating for concentration, removing ethanol, and vacuum freeze drying to obtain Malus micromalus makino extract.
The preparation method of the pumpkin seed extract comprises the following steps: drying defatted pumpkin seeds at a constant temperature at a low temperature, grinding, sieving with a 80-100-mesh sieve to obtain pumpkin seed powder, mixing the pumpkin seed powder with 70% ethanol solution by volume ratio of 1:20, performing reflux extraction at 80 ℃ for 120min, centrifuging and standing after extraction to obtain supernatant, performing rotary evaporation and concentration on the supernatant to remove ethanol, and performing vacuum freeze drying to obtain the pumpkin seed extract.
The preparation method of the grape seed extract comprises the following steps: drying grape seeds at low temperature and constant temperature, crushing and sieving with a 80-100 mesh sieve to obtain grape seed powder, mixing the grape seed powder with an ethanol solution with the volume fraction of 60% according to the solid-to-liquid ratio of 1:25, performing microwave treatment for 120-180 s at the microwave frequency of 2450MHz and the power of 450W, filtering to obtain an extracting solution, performing rotary evaporation and concentration on the extracting solution to remove ethanol, and performing vacuum freeze drying to obtain the grape seed extract.
The preparation method of the blueberry extract comprises the following steps: cleaning fresh blueberries, removing impurities, preparing blueberry pulp, carrying out vacuum freeze drying on the obtained pulp, crushing the obtained pulp, sieving the crushed pulp with a sieve of 80-100 meshes to obtain blueberry powder, mixing the blueberry seed powder with an extraction solution according to a solid-to-liquid ratio of 1:30, carrying out microwave treatment for 240-360 s at a microwave frequency of 2450MHz and a power of 450W, filtering out an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a blueberry extract.
Preferably, the pine pollen is pulverized at the rotating speed of 25000-.
Preferably, the adjuvant is selected from at least one of a filler, a disintegrant, a lubricant, a suspending agent, a binder, a sweetener, a flavoring agent, and a preservative.
Wherein, the filler includes: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.;
the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, cross-linked sodium carboxymethyl cellulose, etc.;
the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like;
the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like;
the adhesive comprises starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.;
the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, and the like;
the flavoring agent comprises: sweeteners and various essences;
the preservative comprises: nipagin, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chloroacetidine acetate, eucalyptus oil, etc
The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.;
the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, cross-linked sodium carboxymethyl cellulose, etc.;
preferably, the dosage form of the composition is an oral preparation, and the oral preparation comprises: one or more of tablet, powder, capsule, soft capsule, pill, granule and oral liquid.
An NMN-containing composition for delaying aging, comprising the steps of:
1) preparing crab apple extract, pumpkin seed extract, grape seed extract, blueberry extract and wall-broken pine pollen;
2) weighing the crab apple extract, the pumpkin seed extract, the wall-broken pollen pini, the grape seed extract and the blueberry extract according to the weight ratio, and stirring and mixing the raw material powder for 5-20min to obtain the slow-mixing uniform composition.
An anti-aging composition containing NMN can be used for preparing food and health product for delaying aging.
Has the advantages that:
1. the invention provides a NMN-containing composition capable of delaying senescence, which takes MN, a crab apple extract, a pumpkin seed extract, wall-broken pine pollen, a grape seed extract and a blueberry extract as main raw materials; one of the important precursors of the NMN mammal NAD + has better treatment and repair effects on cardiovascular and cerebrovascular diseases, neurodegenerative diseases, aging degenerative diseases and the like by regulating the level of NMN in a living body, and the crab apple extract, the pumpkin seed extract, the wall-broken pine pollen, the grape seed extract and the blueberry extract have good antioxidant functions, can remove free radicals and have obvious effect on delaying senescence;
2. according to the preparation method provided by the invention, the raw material components are directly prepared into powder and then mixed and stirred to obtain the composition, so that the problem that the stability of NMN in the composition is damaged by the existing preparation method is avoided, and therefore, the prepared composition containing the NMN has high stability, is convenient to store and transport, and has high dissolution rate, good absorption effect for a human body and high content of effective components;
3. the composition prepared by the preparation method can effectively improve the activity of superoxide dismutase-SOD of mice, obviously improve the activity level of peroxidase-CAT and reduced glutathione-GSH, and reduce the content of malondialdehyde-MDA.
Drawings
FIG. 1 shows the effect of the composition reagents on the activity of superoxide dismutase in the serum, brain tissue, liver, kidney and spleen of D-galactose subacute aging mice;
FIG. 2 is a graph showing the effect of the composition reagents on peroxidase activity in serum, brain tissue and liver, kidney and spleen of D-galactose subacute aging mice;
FIG. 3 is a graph showing the effect of the composition reagents on reduced glutathione activity in serum, brain tissue and liver, kidney, spleen of D-galactose subacute aging mice;
FIG. 4 shows the effect of the composition on the serum, brain tissue and the content of malondialdehyde in the liver, kidney and spleen of D-galactose subacute aging mice.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention, and that various changes or modifications may be suggested to one skilled in the art after reading the teachings herein, and that such equivalents are within the scope of the appended claims.
In the description of the present invention, it is to be understood that the terms "first", "second" and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
The weight of the related components mentioned in the description of the embodiments of the present invention may not only refer to the specific content of each component, but also represent the proportional relationship of the weight among the components, and therefore, the content of the related components is scaled up or down within the scope disclosed in the description of the embodiments of the present invention as long as it is in accordance with the description of the embodiments of the present invention. Specifically, the weight described in the description of the embodiment of the present invention may be a unit of mass known in the chemical industry field, such as μ g, mg, g, and kg.
When the composition of the invention is added with proper auxiliary materials to be prepared into tablets, powder, capsules, soft capsules, pills, granules and oral liquid, the components can be fully stirred, added with proper amount of filling agent and adhesive to be prepared into soft materials, and then granulated to obtain granules; directly encapsulating the granule or directly encapsulating the concentrated extract powder to obtain capsule; adding a certain amount of glidant into the obtained granules, and pressing the granules into tablets to obtain tablets and the like;
the various dosage forms described above are readily prepared by those skilled in the art without requiring special conditions for ordinary formulation.
The present invention will be described in more detail with reference to examples. It is to be understood that the practice of the invention is not limited to the following examples, and that any variations and/or modifications may be made thereto without departing from the scope of the invention.
In the invention, all parts and percentages are weight units, and all equipment, raw materials and the like can be purchased from the market or are commonly used in the industry, if not specified. The methods in the following examples are conventional in the art unless otherwise specified.
Example 1
Preparation of the compositions of the invention
The composition comprises the following raw materials in parts by weight: 5 parts of NMN, 15 parts of crab apple extract, 15 parts of pumpkin seed extract, 3 parts of wall-broken pine pollen, 12 parts of grape seed extract sugar and 12 parts of blueberry extract.
Wherein, the preparation method of the malus micromalus makino extract, the pumpkin seed extract, the grape seed extract and the blueberry extract comprises the following steps:
the preparation method of the malus micromalus makino extract comprises the following steps: cleaning fresh Malus micromalus makino, naturally drying in the shade, removing core, collecting pulp, drying the collected fruit residue at low temperature and constant temperature, pulverizing, sieving with 80-100 mesh sieve, mixing Malus micromalus makino powder with 40% volume fraction ethanol solution according to a solid-to-liquid ratio of 1:35, performing microwave treatment at microwave frequency of 2450MHz and power of 550W for 180-240s, filtering to obtain extractive solution, rotary evaporating for concentration, removing ethanol, and vacuum freeze drying to obtain Malus micromalus makino extract.
The preparation method of the pumpkin seed extract comprises the following steps: drying defatted pumpkin seeds at a constant temperature at a low temperature, grinding, sieving with a 80-100-mesh sieve to obtain pumpkin seed powder, mixing the pumpkin seed powder with 70% ethanol solution by volume ratio of 1:20, performing reflux extraction at 80 ℃ for 120min, centrifuging and standing after extraction to obtain supernatant, performing rotary evaporation and concentration on the supernatant to remove ethanol, and performing vacuum freeze drying to obtain the pumpkin seed extract.
The preparation method of the grape seed extract comprises the following steps: drying grape seeds at low temperature and constant temperature, crushing and sieving with a 80-100 mesh sieve to obtain grape seed powder, mixing the grape seed powder with an ethanol solution with the volume fraction of 60% according to the solid-to-liquid ratio of 1:25, performing microwave treatment for 120-180 s at the microwave frequency of 2450MHz and the power of 450W, filtering to obtain an extracting solution, performing rotary evaporation and concentration on the extracting solution to remove ethanol, and performing vacuum freeze drying to obtain the grape seed extract.
The preparation method of the blueberry extract comprises the following steps: cleaning fresh blueberries, removing impurities, preparing blueberry pulp, carrying out vacuum freeze drying on the obtained pulp, crushing the obtained pulp, sieving the crushed pulp with a sieve of 80-100 meshes to obtain blueberry powder, mixing the blueberry seed powder with an extraction solution according to a solid-to-liquid ratio of 1:30, carrying out microwave treatment for 240-360 s at a microwave frequency of 2450MHz and a power of 450W, filtering out an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a blueberry extract.
Weighing the crab apple extract, the pumpkin seed extract, the wall-broken pollen pini, the grape seed extract and the blueberry extract obtained in the steps according to the weight ratio, and stirring and mixing the raw material powder for 12min to obtain the slow-mixing uniform composition.
Example 2
Preparation of the compositions of the invention
The composition comprises the following raw materials in parts by weight: NMN1 parts, crab apple extract 10 parts, pumpkin seed extract 10 parts, wall-broken pine pollen 2 parts, grape seed extract sugar 10 parts, and blueberry extract 10 parts.
Wherein, the preparation method of the malus micromalus makino extract, the pumpkin seed extract, the grape seed extract and the blueberry extract comprises the following steps:
the preparation method of the malus micromalus makino extract comprises the following steps: cleaning fresh Malus micromalus makino, naturally drying in the shade, removing core, collecting pulp, drying the collected fruit residue at low temperature and constant temperature, pulverizing, sieving with 80-100 mesh sieve, mixing Malus micromalus makino powder with 40% volume fraction ethanol solution according to a solid-to-liquid ratio of 1:35, performing microwave treatment at microwave frequency of 2450MHz and power of 550W for 180-240s, filtering to obtain extractive solution, rotary evaporating for concentration, removing ethanol, and vacuum freeze drying to obtain Malus micromalus makino extract.
The preparation method of the pumpkin seed extract comprises the following steps: drying defatted pumpkin seeds at a constant temperature at a low temperature, grinding, sieving with a 80-100-mesh sieve to obtain pumpkin seed powder, mixing the pumpkin seed powder with 70% ethanol solution by volume ratio of 1:20, performing reflux extraction at 80 ℃ for 120min, centrifuging and standing after extraction to obtain supernatant, performing rotary evaporation and concentration on the supernatant to remove ethanol, and performing vacuum freeze drying to obtain the pumpkin seed extract.
The preparation method of the grape seed extract comprises the following steps: drying grape seeds at low temperature and constant temperature, crushing and sieving with a 80-100 mesh sieve to obtain grape seed powder, mixing the grape seed powder with an ethanol solution with the volume fraction of 60% according to the solid-to-liquid ratio of 1:25, performing microwave treatment for 120-180 s at the microwave frequency of 2450MHz and the power of 450W, filtering to obtain an extracting solution, performing rotary evaporation and concentration on the extracting solution to remove ethanol, and performing vacuum freeze drying to obtain the grape seed extract.
The preparation method of the blueberry extract comprises the following steps: cleaning fresh blueberries, removing impurities, preparing blueberry pulp, carrying out vacuum freeze drying on the obtained pulp, crushing the obtained pulp, sieving the crushed pulp with a sieve of 80-100 meshes to obtain blueberry powder, mixing the blueberry seed powder with an extraction solution according to a solid-to-liquid ratio of 1:30, carrying out microwave treatment for 240-360 s at a microwave frequency of 2450MHz and a power of 450W, filtering out an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a blueberry extract.
Weighing the crab apple extract, the pumpkin seed extract, the wall-broken pollen pini, the grape seed extract and the blueberry extract obtained in the steps according to the weight ratio, and stirring and mixing the raw material powder for 5min to obtain the slow-mixing uniform composition.
Example 3
Preparation of the compositions of the invention
The composition comprises the following raw materials in parts by weight: NMN10 parts, resveratrol 10 parts, angelica polysaccharide 20 parts, lophatherum gracile polysaccharide 20 parts, codonopsis pilosula polysaccharide 5 parts, wolfberry polysaccharide 25 parts, cordyceps sinensis extract 20 parts, ginseng extract 20 parts, pilose antler extract 20 parts, fleece-flower root extract 12 parts, rhodiola rosea extract 10 parts, epimedium extract 20 parts and vitamin E10 parts.
Wherein, the preparation method of the malus micromalus makino extract, the pumpkin seed extract, the grape seed extract and the blueberry extract comprises the following steps:
the preparation method of the malus micromalus makino extract comprises the following steps: cleaning fresh Malus micromalus makino, naturally drying in the shade, removing core, collecting pulp, drying the collected fruit residue at low temperature and constant temperature, pulverizing, sieving with 80-100 mesh sieve, mixing Malus micromalus makino powder with 40% volume fraction ethanol solution according to a solid-to-liquid ratio of 1:35, performing microwave treatment at microwave frequency of 2450MHz and power of 550W for 180-240s, filtering to obtain extractive solution, rotary evaporating for concentration, removing ethanol, and vacuum freeze drying to obtain Malus micromalus makino extract.
The preparation method of the pumpkin seed extract comprises the following steps: drying defatted pumpkin seeds at a constant temperature at a low temperature, grinding, sieving with a 80-100-mesh sieve to obtain pumpkin seed powder, mixing the pumpkin seed powder with 70% ethanol solution by volume ratio of 1:20, performing reflux extraction at 80 ℃ for 120min, centrifuging and standing after extraction to obtain supernatant, performing rotary evaporation and concentration on the supernatant to remove ethanol, and performing vacuum freeze drying to obtain the pumpkin seed extract.
The preparation method of the grape seed extract comprises the following steps: drying grape seeds at low temperature and constant temperature, crushing and sieving with a 80-100 mesh sieve to obtain grape seed powder, mixing the grape seed powder with an ethanol solution with the volume fraction of 60% according to the solid-to-liquid ratio of 1:25, performing microwave treatment for 120-180 s at the microwave frequency of 2450MHz and the power of 450W, filtering to obtain an extracting solution, performing rotary evaporation and concentration on the extracting solution to remove ethanol, and performing vacuum freeze drying to obtain the grape seed extract.
The preparation method of the blueberry extract comprises the following steps: cleaning fresh blueberries, removing impurities, preparing blueberry pulp, carrying out vacuum freeze drying on the obtained pulp, crushing the obtained pulp, sieving the crushed pulp with a sieve of 80-100 meshes to obtain blueberry powder, mixing the blueberry seed powder with an extraction solution according to a solid-to-liquid ratio of 1:30, carrying out microwave treatment for 240-360 s at a microwave frequency of 2450MHz and a power of 450W, filtering out an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a blueberry extract.
Weighing the crab apple extract, the pumpkin seed extract, the wall-broken pollen pini, the grape seed extract and the blueberry extract obtained in the steps according to the weight ratio, and stirring and mixing the raw material powder for 20min to obtain the slow-mixing uniform composition.
Examples of the experiments
The experimental examples required drugs and reagents:
d-galactose: purchased from Nanjing Kangkui chemical reagents, Inc.;
superoxide dismutase (SOD), Malondialdehyde (MDA), reduced Glutathione (GSH), peroxidase (CAT) kit: purchased from Nanjing to build a bioengineering institute;
the composition of the invention: example 1
Experimental animals and feeding conditions:
60 male Kunming mice, aged two months, of clean grade and body weight (22 +/-2) g, are provided by the center of the laboratory animal of the Sichuan antibiotic research institute;
feeding conditions are as follows: test animal house is SPF grade, temperature: 20-25 deg.C, 40-70% relative humidity, and is fed with normal drinking water and diet.
The experimental method comprises the following steps:
1. experimental grouping and processing
After 7 days of adaptive feeding, mice were randomized into three groups: a normal control group, a D-galactose aging group and an experimental group, wherein each group comprises 20 animals, and the experimental group adopts the composition of the invention to victory saline; wherein D-galactose is injected into abdominal cavity of the aging group and the experimental group according to the dosage of 150 mg/(kg. D), and 0.9% sterile physiological salt with the same amount is injected into the normal control group, and a subacute aging model is established for 42 days continuously; performing intragastric intervention according to the body weight after intraperitoneal injection every day by referring to pharmacological intragastric dose (0.1mL/10g), and performing intragastric administration once every day, wherein the normal control group and the D-galactose aging group are both intragastric administered with 0.9% sterile normal saline; the test group is filled with 100mg/kg of stomach, the weight of the mouse is weighed and recorded every week, and the stomach filling dosage is adjusted for 42 days continuously; fasting is carried out for 12 hours after the last administration, cervical vertebra dislocation is killed, blood is taken from eyeball veins, serum is sucked for standby, meanwhile, brain, liver, kidney, thymus, spleen and the like are removed, after ice physiological saline is cleaned, the weight of the thymus and the spleen is weighed by an electronic balance, and brain tissues, the serum and part of the liver are stored at the temperature of minus 80 ℃ for subsequent experiments.
2. Behavioural testing
Weight changes of the mice were recorded during the administration, and changes in the vitality and morphology of the mice were observed.
3. Biochemical index detection
3.1 thymus and spleen indices calculation using the following formula:
mouse thymus index-weight thymus (mg)/weight (g);
mouse spleen index ═ spleen weight (mg)/body weight (g);
3.2 determination of serum Biochemical indicators of laboratory animals
After the last administration, blood is taken from the eyeball plexus vein, the blood is placed in a centrifuge tube and centrifuged for 10min at 4200r/min at 4 ℃ to obtain serum, and the activity of superoxide dismutase-SOD, the activity of peroxidase-CAT, the activity of reduced glutathione-GSH and the content of malonaldehyde-MDA in the serum of the mouse are respectively measured according to the using instruction of the kit.
3.3 biochemical index determination of brain tissue and partial liver
Weighing about 50mg (wet weight) of each tissue sample of the mouse, adding 450 mu L of precooled 0.9% sterile physiological saline, and preparing 10% homogenate by a homogenizer; then centrifuging at 4000r/min for 10min at 4 ℃ to obtain supernatant of each tissue homogenate. The activity of superoxide dismutase-SOD, the activity of peroxidase-CAT, the activity of reduced glutathione-GSH and the content of malondialdehyde-MDA in brain tissues and livers of mice are respectively measured according to the operation of the instruction of the kit.
4. Data processing
All experimental data were analyzed using SPSS 18.0 for Window software and expressed as (x. + -.s) using one-way analysis of variance, and if no significant differences between groups were noted, the groups were analyzed for pairwise comparisons using LSD, and if the variance was not uniform, the Dunnett's method was used.
Results and comparison
TABLE 1 behavioral testing of groups of mice
Note that + indicates cachexia, significantly reduced activity, easy grasping, and a significant aging status; + indicates a poor spirit and a decreased activity; + + + + indicates good spirit and normal activity; + + + + + indicates normal spirit, vigorous activity and not easy to grab; (M)1-M2) Represents the difference in body weight; (M)1-M2)/M2The weight gain is shown.
As can be seen from Table 1, at the end of the experiment, the mice in the D-galactose aging model group had an aging state, which is mainly characterized by dark hair, lassitude, weakness of limbs, reduced activity, relatively small vitality felt during grabbing, easy grabbing, and reliable modeling of the aging model mice; the weight of the mice in the D-galactose aging model group increased slowly compared to the normal control group, and the experimental group fed with the composition physiological saline solution promoted the slow weight increase of the mice in the D-galactose aging model group to be higher than that in the galactose aging model group, but the increase was still lower than that in the normal control group.
TABLE 2 changes in thymus index and spleen index in each group of mice
| Group of | Thymus index (mg/g) | Spleen index (mg/g) |
| Normal control group | 1.48±0.26 | 3.27±0.95 |
| D-galactose senescence group | 1.13±0.1911) | 2.05±0.4311) |
| Experimental group | 1.40±0.1122) | 3.19±0.2522) |
Note: comparison with normal control group: ) P < 0.05; comparison with model groups: 2) p < 0.05;
thymus and spleen are important immune organs of the body and show more obvious change-degeneration phenomenon along with age than other organs, namely the body mass index of the thymus and the spleen is reduced, so that the thymus and the spleen are atrophied and regarded as the symptoms of the body aging; as can be seen from Table 2, the thymus index and spleen index of the D-galactose-senescent group were both significantly lower than those of the control group (P < 0.05); the thymus index and spleen index of the experimental group are obviously higher than those of the model group (P <0.05), experiments show that the composition has the weight-gaining effect on thymus and spleen of immune organs of mice, and the composition of the experimental group has the effect of delaying thymus and spleen wilting aging.
GSH is one of important antioxidants and free radical scavengers in vivo, can effectively relieve oxidative damage in organisms, and the content of GSH can measure an important index of the oxidation resistance of the organisms; MDA is a product of lipid peroxidation initiated by free radicals in the body and is commonly used to assess the degree of aging; SOD is an important antioxidant enzyme in vivo, can effectively remove superoxide anion free radicals, and generates hydrogen peroxide; CAT can decompose hydrogen peroxide, and can cooperate with SOD to completely remove superoxide ions, reduce MDA and free radical metabolites, and prevent cells from being broken;
as shown in fig. 1-4, compared with the normal control group, the activities of superoxide dismutase-SOD, peroxidase-CAT and reduced glutathione-GSH in serum, brain tissue and liver of the D-galactose aging group are significantly reduced, and the malondialdehyde-MDA content is significantly increased; compared with the D-galactose aging group, the activity of superoxide dismutase-SOD, the activity of peroxidase-CAT and the activity level of reduced glutathione-GSH in serum, brain tissues and liver in the experimental group are obviously improved, the malondialdehyde-MDA content is reduced, and statistical difference exists; the results show that the composition of the invention can enhance the activity of antioxidant enzyme in serum and organs, inhibit the growth of free radicals by improving the activity of free radical scavengers in the body, reduce lipid peroxidation and protect tissues from the attack of oxidants and free radicals, thereby playing the role of anti-aging.
Claims (10)
1. An NMN-containing composition for retarding aging, comprising: the composition comprises main materials and auxiliary materials, wherein the main materials of the composition comprise: NMN, crab apple extract, pumpkin seed extract, wall-broken pine pollen, grape seed extract and blueberry extract.
2. The NMN-containing senescence delaying composition according to claim 1, wherein: the main material of the composition comprises the following raw materials in parts by weight: NMN1-10 parts, crab apple extract 10-20 parts, pumpkin seed extract 10-20 parts, wall-broken pine pollen 2-5 parts, grape seed extract sugar 10-15 parts, and blueberry extract 10-15 parts.
3. The NMN-containing senescence delaying composition according to claim 2, wherein: the main material of the composition comprises the following raw materials in parts by weight: 5 parts of NMN, 15 parts of crab apple extract, 15 parts of pumpkin seed extract, 3 parts of wall-broken pine pollen, 12 parts of grape seed extract sugar and 12 parts of blueberry extract.
4. A NMN-containing senescence delaying composition according to any one of claims 1-3, wherein: the preparation method of the malus micromalus makino extract, the pumpkin seed extract, the grape seed extract and the blueberry extract comprises the following steps:
the preparation method of the malus micromalus makino extract comprises the following steps: cleaning fresh Malus micromalus makino, naturally drying in the shade, removing core, taking pulp part, drying the collected fruit residue at low temperature and constant temperature, then crushing and sieving by a 80-100 mesh sieve, mixing Malus micromalus makino powder with 40% volume fraction ethanol solution according to a solid-liquid ratio of 1:35, performing microwave treatment for 180-240s at microwave frequency of 2450MHz and power of 550W, filtering to obtain an extracting solution, performing rotary evaporation and concentration on the extracting solution, removing ethanol, and performing vacuum freeze drying to obtain Malus micromalus makino extract;
the preparation method of the pumpkin seed extract comprises the following steps: drying defatted pumpkin seeds at a constant temperature at a low temperature, grinding, sieving with a 80-100-mesh sieve to obtain pumpkin seed powder, mixing the pumpkin seed powder with 70% ethanol solution by volume fraction according to a solid-to-liquid ratio of 1:20, performing reflux extraction at 80 ℃ for 120min, standing in a centrifuge after extraction to obtain supernatant, performing rotary evaporation and concentration on the supernatant to remove ethanol, and performing vacuum freeze drying to obtain a pumpkin seed extract;
the preparation method of the grape seed extract comprises the following steps: drying grape seeds at a low temperature and a constant temperature, crushing and sieving the grape seeds with a sieve of 80-100 meshes to obtain grape seed powder, mixing the grape seed powder with an ethanol solution with a volume fraction of 60% according to a solid-to-liquid ratio of 1:25, carrying out microwave treatment for 120-180 s at a microwave frequency of 2450MHz and a power of 450W, filtering an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a grape seed extract;
the preparation method of the blueberry extract comprises the following steps: cleaning fresh blueberries, removing impurities, preparing blueberry pulp, carrying out vacuum freeze drying on the obtained pulp, crushing the obtained pulp, sieving the crushed pulp with a sieve of 80-100 meshes to obtain blueberry powder, mixing the blueberry seed powder with an extraction solution according to a solid-to-liquid ratio of 1:30, carrying out microwave treatment for 240-360 s at a microwave frequency of 2450MHz and a power of 450W, filtering out an extracting solution, carrying out rotary evaporation and concentration on the extracting solution to remove ethanol, and carrying out vacuum freeze drying to obtain a blueberry extract.
5. A NMN-containing senescence delaying composition according to any one of claims 1-3, wherein: the preparation method of the pine pollen wall-broken comprises the following steps: pulverizing pollen Pini at 25000-.
6. The NMN-containing senescence delaying composition according to claim 1, wherein: the adjuvant is selected from at least one of bulking agent, disintegrating agent, lubricant, suspending agent, binder, sweetener, correctant and antiseptic.
7. The NMN-containing senescence delaying composition according to claim 1, wherein: the dosage form of the composition is an oral preparation, and the oral preparation comprises: one or more of tablet, powder, capsule, soft capsule, pill, granule and oral liquid.
8. A method for preparing a composition containing NMN for delaying aging, comprising the steps of: the method comprises the following steps:
1) preparing crab apple extract, pumpkin seed extract, grape seed extract, blueberry extract and wall-broken pine pollen;
2) weighing the crab apple extract, the pumpkin seed extract, the wall-broken pollen pini, the grape seed extract and the blueberry extract according to the weight ratio, and stirring and mixing the raw material powder for 5-20min to obtain the slow-mixing uniform composition.
9. The method for preparing an NMN-containing senescence delaying composition according to claim 8, wherein: and 2) adding related auxiliary materials, uniformly mixing, and preparing one or more of tablets, powder, capsules, soft capsules, pills, granules and oral liquid.
10. Use of NMN-containing senescence delaying compositions according to claim 1, wherein: the composition can be used for preparing food and health care products for delaying senility.
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| CN115010772A (en) * | 2022-06-16 | 2022-09-06 | 国珍健康科技（北京）有限公司 | Nicotinamide mononucleotide-containing pine pollen extract, extract and preparation method thereof |
| WO2024026740A1 (en) * | 2022-08-03 | 2024-02-08 | 大汉酵素生物科技股份有限公司 | Fermentation composition with nmn anti-aging effect |
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